From dd50cf58d9651c5e72f0d76ba3300b8dff6ecbcd Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 24 Jan 2025 11:57:52 +0100 Subject: [PATCH 001/317] Update main.nf to trigger tests From 8505da9dceea5e9305f0c58ff5cac75ff248a442 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 24 Jan 2025 12:00:47 +0100 Subject: [PATCH 002/317] Update test.config -> 8 GB RAM --- conf/test.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test.config b/conf/test.config index 8c566af..90aff90 100644 --- a/conf/test.config +++ b/conf/test.config @@ -13,7 +13,7 @@ process { resourceLimits = [ cpus: 4, - memory: '15.GB', + memory: '8.GB', time: '1.h' ] } From dcbc7adc694058a1b754a2bf04749e5ec2b04259 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Fri, 24 Jan 2025 14:21:42 +0100 Subject: [PATCH 003/317] samplesheet.csv, schema_input.json, subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf --- assets/samplesheet.csv | 13 ++++-- assets/schema_input.json | 42 +++++++++++++---- .../utils_nfcore_dmscore_pipeline/main.nf | 46 ++++++++++++------- 3 files changed, 72 insertions(+), 29 deletions(-) diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv index 5f653ab..7477367 100644 --- a/assets/samplesheet.csv +++ b/assets/samplesheet.csv @@ -1,3 +1,10 @@ -sample,fastq_1,fastq_2 -SAMPLE_PAIRED_END,/path/to/fastq/files/AEG588A1_S1_L002_R1_001.fastq.gz,/path/to/fastq/files/AEG588A1_S1_L002_R2_001.fastq.gz -SAMPLE_SINGLE_END,/path/to/fastq/files/AEG588A4_S4_L003_R1_001.fastq.gz, +sample,type,replicate,file1,file2 +a_1,input,1,/path1/forward.fastq,/path1/reverse.fastq +a_1,input,2,/path2/forward.fastq,/path2/reverse.fastq +a_1,output,1,/path3/blabla_se.bam, +a_3,input,1,/path4/forward.fastq,/path4/reverse.fastq +a_3,input,2,/path5/forward.fastq,/path5/reverse.fastq +a_3,output,1,/path6/forward.fastq,/path6/reverse.fastq +a_3,output,2,/path7/forward.fastq,/path7/reverse.fastq +b_1,quality,1,/path8/single_end.fastq,/path8/reverse.fastq +b_1,quality,2,/path9/forward.fastq,/path9/reverse.fastq diff --git a/assets/schema_input.json b/assets/schema_input.json index 2d9d8f6..8ceec99 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -9,25 +9,47 @@ "properties": { "sample": { "type": "string", - "pattern": "^\\S+$", - "errorMessage": "Sample name must be provided and cannot contain spaces", + "pattern": "^[^\\s/]+$", + "errorMessage": "Sample name must be provided, cannot contain spaces, and must not include special characters", "meta": ["id"] }, - "fastq_1": { + "file1": { "type": "string", "format": "file-path", "exists": true, - "pattern": "^\\S+\\.f(ast)?q\\.gz$", - "errorMessage": "FastQ file for reads 1 must be provided, cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'" + "pattern": "^\\S+\\.(bam|fastq|fq|fastq\\.gz|fq\\.gz)$", + "allOf": [ + { + "if": { + "pattern": "^\\S+\\.bam$" + }, + "then": { + "pattern": "^\\S+_(pe|se)\\.bam$", + "errorMessage": "If file1 ends with .bam, it must contain '_pe.bam' or '_se.bam', defining paired-end or single-end" + } + } + ], + "errorMessage": "File 1 must be provided, cannot contain spaces, and must have an allowed extension (.bam, .fastq, .fq, .fastq.gz, .fq.gz)" }, - "fastq_2": { - "type": "string", + "file2": { + "type": ["string", "null"], "format": "file-path", "exists": true, - "pattern": "^\\S+\\.f(ast)?q\\.gz$", - "errorMessage": "FastQ file for reads 2 cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'" + "pattern": "^\\S+\\.(fastq|fq|fastq\\.gz|fq\\.gz)$", + "errorMessage": "File 2 must have an allowed extension (.fastq, .fq, .fastq.gz, .fq.gz) or be null for single-end reads" + }, + "type": { + "type": "string", + "enum": ["input", "output", "quality"], + "errorMessage": "Type must be one of: input, output, or quality" + }, + "replicate": { + "type": "integer", + "minimum": 1, + "errorMessage": "Replicate must be a positive integer" } }, - "required": ["sample", "fastq_1"] + "required": ["sample", "file1", "type", "replicate"] } } + diff --git a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf b/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf index 14eba3c..512ab2d 100644 --- a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf @@ -73,25 +73,39 @@ workflow PIPELINE_INITIALISATION { // Channel - .fromList(samplesheetToList(params.input, "${projectDir}/assets/schema_input.json")) - .map { - meta, fastq_1, fastq_2 -> - if (!fastq_2) { - return [ meta.id, meta + [ single_end:true ], [ fastq_1 ] ] - } else { - return [ meta.id, meta + [ single_end:false ], [ fastq_1, fastq_2 ] ] - } - } - .groupTuple() - .map { samplesheet -> - validateInputSamplesheet(samplesheet) + .fromPath(params.input) + .splitCsv(header: true) + .filter { row -> + // Skip rows where file1 or file2 are BAM files + !(row.file1.endsWith('.bam') || (row.file2 && row.file2.endsWith('.bam'))) + } + .map { row -> + // Determine suffix based on the presence of file2 + def suffix = row.file2 ? "_pe" : "_se" + + // Construct metadata object with updated ID + def meta = [ + id : "${row.sample}_${row.type}_${row.replicate}${suffix}", // Base ID with suffix + sample : row.sample, + type : row.type, + replicate : row.replicate as int + ] + + // Generate file paths based on the presence of file1 and file2 + def reads = [] + if (row.file1) { + reads << row.file1 // Add file1 path } - .map { - meta, fastqs -> - return [ meta, fastqs.flatten() ] + if (row.file2) { + reads << row.file2 // Add file2 path } - .set { ch_samplesheet } + // Return metadata and file paths as a tuple + return [meta, reads] + } + .set { ch_samplesheet } + +// Emit the samplesheet channel and an empty version channel for use in the workflow emit: samplesheet = ch_samplesheet versions = ch_versions From 02aac6f1cc6e008fd5874304a4121c6793a7dc00 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Fri, 24 Jan 2025 14:30:54 +0100 Subject: [PATCH 004/317] applied prettier on schema_input.json --- assets/schema_input.json | 1 - 1 file changed, 1 deletion(-) diff --git a/assets/schema_input.json b/assets/schema_input.json index 8ceec99..a0984ec 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -52,4 +52,3 @@ "required": ["sample", "file1", "type", "replicate"] } } - From 05457794b639931a8bfadb6b2b9ea38d40d3e98b Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 24 Jan 2025 14:41:50 +0100 Subject: [PATCH 005/317] Update ci.yml --- .github/workflows/ci.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 92396ce..fb58f6d 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -22,7 +22,7 @@ jobs: test: name: "Run pipeline with test data (${{ matrix.NXF_VER }} | ${{ matrix.test_name }} | ${{ matrix.profile }})" # Only run on push if this is the nf-core dev branch (merged PRs) - if: "${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/dmscore') }}" + if: "${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'benjaminwehnert1008/dmscore') }}" runs-on: ubuntu-latest strategy: matrix: From fc742370d916bbe4c6b42a1e18b3e986be9bc802 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Fri, 24 Jan 2025 14:52:53 +0100 Subject: [PATCH 006/317] testdata paths --- nextflow.config | 2 +- nextflow_schema.json | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/nextflow.config b/nextflow.config index 3fd3e96..fc3c7d2 100644 --- a/nextflow.config +++ b/nextflow.config @@ -37,7 +37,7 @@ params { help_full = false show_hidden = false version = false - pipelines_testdata_base_path = 'https://raw.githubusercontent.com/nf-core/test-datasets/' + pipelines_testdata_base_path = 'https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/' trace_report_suffix = new java.util.Date().format( 'yyyy-MM-dd_HH-mm-ss')// Config options config_profile_name = null config_profile_description = null diff --git a/nextflow_schema.json b/nextflow_schema.json index e4e5523..2103634 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -216,7 +216,7 @@ "type": "string", "fa_icon": "far fa-check-circle", "description": "Base URL or local path to location of pipeline test dataset files", - "default": "https://raw.githubusercontent.com/nf-core/test-datasets/", + "default": "https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/", "hidden": true }, "trace_report_suffix": { From 2a13f1b76dda8a1bc6dcd58cc95e001aa419d27d Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 24 Jan 2025 15:04:30 +0100 Subject: [PATCH 007/317] Update test.config --- conf/test.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test.config b/conf/test.config index 90aff90..6871e67 100644 --- a/conf/test.config +++ b/conf/test.config @@ -25,6 +25,6 @@ params { // Input data // TODO nf-core: Specify the paths to your test data on nf-core/test-datasets // TODO nf-core: Give any required params for the test so that command line flags are not needed - input = params.pipelines_testdata_base_path + 'viralrecon/samplesheet/samplesheet_test_illumina_amplicon.csv'// Genome references + input = params.pipelines_testdata_base_path + 'samplesheet_qc_only.csv'// Genome references genome = 'R64-1-1' } From 5cc5cdc9b2e10c7483c161a1674d77d33b247a23 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 24 Jan 2025 15:04:56 +0100 Subject: [PATCH 008/317] Update test_full.config --- conf/test_full.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test_full.config b/conf/test_full.config index b57a75c..6bc2264 100644 --- a/conf/test_full.config +++ b/conf/test_full.config @@ -17,7 +17,7 @@ params { // Input data for full size test // TODO nf-core: Specify the paths to your full test data ( on nf-core/test-datasets or directly in repositories, e.g. SRA) // TODO nf-core: Give any required params for the test so that command line flags are not needed - input = params.pipelines_testdata_base_path + 'viralrecon/samplesheet/samplesheet_full_illumina_amplicon.csv' + input = params.pipelines_testdata_base_path + 'samplesheet_qc_only.csv' // Genome references genome = 'R64-1-1' From 1b385066d01c1ccb239e8afde4e806f821be5fc3 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Fri, 24 Jan 2025 15:21:09 +0100 Subject: [PATCH 009/317] Fix logo to match nf-core template --- docs/images/nf-core-dmscore_logo_dark.png | Bin 28505 -> 25810 bytes 1 file changed, 0 insertions(+), 0 deletions(-) diff --git a/docs/images/nf-core-dmscore_logo_dark.png b/docs/images/nf-core-dmscore_logo_dark.png index 7cbb4e158582ad1611df87b711fecafbd45ec867..3c849477cd21c29a708c41378961eb1f5aeff47e 100644 GIT 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a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf @@ -44,7 +44,7 @@ workflow PIPELINE_INITIALISATION { version, true, outdir, - workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1 + workflow.profile.tokenize(',').intersect(['docker', 'singularity']).size() >= 1 ) // From 089b91322d4ead917f3f961ecbd43f32ca6a3cda Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 24 Jan 2025 16:07:14 +0100 Subject: [PATCH 012/317] remove previous commit --- subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf b/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf index 9f517c8..512ab2d 100644 --- a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf @@ 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subworkflows/local/utils_nfcore_dmsqc_pipeline/main.nf --- subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf | 5 ----- 1 file changed, 5 deletions(-) diff --git a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf b/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf index 512ab2d..0e9660f 100644 --- a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf @@ -63,11 +63,6 @@ workflow PIPELINE_INITIALISATION { nextflow_cli_args ) - // - // Custom validation for pipeline parameters - // - validateInputParameters() - // // Create channel from input file provided through params.input // From 64f865b35ea80521c3ece735fb18740d11310fe1 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 24 Jan 2025 17:13:31 +0100 Subject: [PATCH 019/317] Update main.nf --- subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf b/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf index 0e9660f..512ab2d 100644 --- a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf @@ -63,6 +63,11 @@ workflow PIPELINE_INITIALISATION { nextflow_cli_args ) + // + // Custom validation for pipeline parameters + // + validateInputParameters() + // // Create channel from input file provided through params.input // From aaa04b1216408fe7c94a592d5bbcb966b6ed79c1 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 24 Jan 2025 17:58:01 +0100 Subject: [PATCH 020/317] Update nextflow.config: changed docker.runOptions to process.containerOptions for docker and arm profiles --- nextflow.config | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/nextflow.config b/nextflow.config index fc3c7d2..256e8ea 100644 --- a/nextflow.config +++ b/nextflow.config @@ -89,10 +89,10 @@ profiles { shifter.enabled = false charliecloud.enabled = false apptainer.enabled = false - docker.runOptions = '-u $(id -u):$(id -g)' + process.containerOptions = '-u $(id -u):$(id -g)' } arm { - docker.runOptions = '-u $(id -u):$(id -g) --platform=linux/amd64' + process.containerOptions = '-u $(id -u):$(id -g) --platform=linux/amd64' } singularity { singularity.enabled = true From 1d57bbdd1453b3b0d77be8f916643a65b2d2543d Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 24 Jan 2025 18:01:53 +0100 Subject: [PATCH 021/317] Update modules.config: add <> to FASTQC --- conf/modules.config | 1 + 1 file changed, 1 insertion(+) diff --git a/conf/modules.config b/conf/modules.config index d203d2b..36edb1d 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -20,6 +20,7 @@ process { withName: FASTQC { ext.args = '--quiet' + containerOptions = '' } withName: 'MULTIQC' { From ce3cd9be29275472cac5754d42490341412836e1 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 28 Jan 2025 09:48:06 +0000 Subject: [PATCH 022/317] Fix linting issue via GitHub Codespaces --- .github/PULL_REQUEST_TEMPLATE.md | 4 ++-- .github/workflows/linting_comment.yml | 2 +- docs/images/nf-core-dmscore_logo_dark.png | Bin 28505 -> 28474 bytes 3 files changed, 3 insertions(+), 3 deletions(-) diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index 0d6f133..13cc97f 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -8,14 +8,14 @@ These are the most common things requested on pull requests (PRs). Remember that PRs should be made against the dev branch, unless you're preparing a pipeline release. -Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/dmscore/tree/master/.github/CONTRIBUTING.md) +Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/dmscore/tree/main/.github/CONTRIBUTING.md) --> ## PR checklist - [ ] This comment contains a description of changes (with reason). - [ ] If you've fixed a bug or added code that should be tested, add tests! -- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/dmscore/tree/master/.github/CONTRIBUTING.md) +- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/dmscore/tree/main/.github/CONTRIBUTING.md) - [ ] If necessary, also make a PR on the nf-core/dmscore _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository. - [ ] Make sure your code lints (`nf-core pipelines lint`). - [ ] Ensure the test suite passes (`nextflow run . -profile test,docker --outdir `). diff --git a/.github/workflows/linting_comment.yml b/.github/workflows/linting_comment.yml index 0bed96d..95b6b6a 100644 --- a/.github/workflows/linting_comment.yml +++ b/.github/workflows/linting_comment.yml @@ -11,7 +11,7 @@ jobs: runs-on: ubuntu-latest steps: - name: Download lint results - uses: dawidd6/action-download-artifact@80620a5d27ce0ae443b965134db88467fc607b43 # v7 + uses: dawidd6/action-download-artifact@20319c5641d495c8a52e688b7dc5fada6c3a9fbc # v8 with: workflow: linting.yml workflow_conclusion: completed diff --git a/docs/images/nf-core-dmscore_logo_dark.png b/docs/images/nf-core-dmscore_logo_dark.png index 7cbb4e158582ad1611df87b711fecafbd45ec867..60abaa1cc0916ff38945395b034f3a5b8367420c 100644 GIT binary patch literal 28474 zcmd42WmlWu6E+&OXmJScUL=Ghb<*^*AR$Y`Ps+Gswp|8Zi^d*Vm-_>Z?bbh?bc 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BenjaminWehnert1008 Date: Tue, 28 Jan 2025 11:09:25 +0100 Subject: [PATCH 023/317] locally: pipelines lint --fix files_unchanged --- .github/PULL_REQUEST_TEMPLATE.md | 4 ++-- .github/workflows/linting_comment.yml | 2 +- docs/images/nf-core-dmscore_logo_dark.png | Bin 28474 -> 28505 bytes 3 files changed, 3 insertions(+), 3 deletions(-) diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index 13cc97f..0d6f133 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -8,14 +8,14 @@ These are the most common things requested on pull requests (PRs). Remember that PRs should be made against the dev branch, unless you're preparing a pipeline release. -Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/dmscore/tree/main/.github/CONTRIBUTING.md) +Learn more about contributing: [CONTRIBUTING.md](https://github.com/nf-core/dmscore/tree/master/.github/CONTRIBUTING.md) --> ## PR checklist - [ ] This comment contains a description of changes (with reason). - [ ] If you've fixed a bug or added code that should be tested, add tests! -- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/dmscore/tree/main/.github/CONTRIBUTING.md) +- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/dmscore/tree/master/.github/CONTRIBUTING.md) - [ ] If necessary, also make a PR on the nf-core/dmscore _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository. - [ ] Make sure your code lints (`nf-core pipelines lint`). - [ ] Ensure the test suite passes (`nextflow run . -profile test,docker --outdir `). diff --git a/.github/workflows/linting_comment.yml b/.github/workflows/linting_comment.yml index 95b6b6a..0bed96d 100644 --- a/.github/workflows/linting_comment.yml +++ b/.github/workflows/linting_comment.yml @@ -11,7 +11,7 @@ jobs: runs-on: ubuntu-latest steps: - name: Download lint results - uses: dawidd6/action-download-artifact@20319c5641d495c8a52e688b7dc5fada6c3a9fbc # v8 + uses: dawidd6/action-download-artifact@80620a5d27ce0ae443b965134db88467fc607b43 # v7 with: workflow: linting.yml workflow_conclusion: completed diff --git a/docs/images/nf-core-dmscore_logo_dark.png b/docs/images/nf-core-dmscore_logo_dark.png index 60abaa1cc0916ff38945395b034f3a5b8367420c..7cbb4e158582ad1611df87b711fecafbd45ec867 100644 GIT binary patch literal 28505 zcmd42bz58A^F182xH}YgC=i_DTHL(^D8XHdyOiP-cPJ9vr4;u-ad&rj*N6M_{k@3i 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zk>PzhlGh^;{G1-fo?CxUvMJ%(>I=<{Jwgh`Lcw(Q>JAk--&aSjB+=ALmhQsCzR^Ke z2F>jU2&@Nw5+TrnCNSKVc*B&L2Jog&yGU*KvNm^oVZtVC!Eleu4+$yze_pORl&qPK zCduM}9$^AU*$I+@gI1r?!-4QSjhum498bfvi7Nk5vr1_`{p1fa!#lx+@a($Bc(LZ_ z9s=0#v|dEuGWopC0KzK#drBBG+RSZ!&UZ;3BS`+;ubFc4vgyvcPjU{dB~n3-;FQN&7lgpQ!HLMIC}L5|Pj05KXNK(b8r z-Ys+~kf_VVjo*<71ldAmueQfgUEOLxwH>>cWxFjHhTvVMb%ojlcU!1nPoiRX&UH$> zvH^H<<)r1Gs?5P-9I*zr^rE{sqSVH zUK)0$?K9$1lSJ3iNvVpjnT;kkeG%B13z-qlD%m}(TFBaa(j2l#b2|y6S>CH@w+2?{{W)P|HJ%Nm-Dn8Ns1J>i57J-g2(ZtJ=^l*(qw$r6O3juTuic8llnQ8 z)xsiex`1VsOExZQR|9Ix%mToZqf19m37QaSK1Ub#Kn!SiTAPV2>J-G@>e9jL6fi}h z#9avB{q$#tlM-)a2?M5eo2L%-bAEb94!FeuG&22UtY~N6gNE1|)ckvJb1FRsJP0Of umaW<`kv=8{RMP)@@$CP{_c9AM#h=P4Sa)qTH~{`K53;g2hpI$+-uyoWHV=yc From d02ece9be4a08a5d53757b9594b60811a15246d2 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Tue, 28 Jan 2025 12:50:53 +0100 Subject: [PATCH 025/317] add bwa index module --- modules.json | 5 ++ modules/nf-core/bwa/index/environment.yml | 5 ++ modules/nf-core/bwa/index/main.nf | 53 +++++++++++++++++++ modules/nf-core/bwa/index/meta.yml | 52 ++++++++++++++++++ modules/nf-core/bwa/index/tests/main.nf.test | 33 ++++++++++++ .../nf-core/bwa/index/tests/main.nf.test.snap | 47 ++++++++++++++++ modules/nf-core/bwa/index/tests/tags.yml | 2 + 7 files changed, 197 insertions(+) create mode 100644 modules/nf-core/bwa/index/environment.yml create mode 100644 modules/nf-core/bwa/index/main.nf create mode 100644 modules/nf-core/bwa/index/meta.yml create mode 100644 modules/nf-core/bwa/index/tests/main.nf.test create mode 100644 modules/nf-core/bwa/index/tests/main.nf.test.snap create mode 100644 modules/nf-core/bwa/index/tests/tags.yml diff --git a/modules.json b/modules.json index 910602b..026d225 100644 --- a/modules.json +++ b/modules.json @@ -5,6 +5,11 @@ "https://github.com/nf-core/modules.git": { "modules": { "nf-core": { + "bwa/index": { + "branch": "master", + "git_sha": "2d20463181b1c38981a02e90d3084b5f9fa8d540", + "installed_by": ["modules"] + }, "fastqc": { "branch": "master", "git_sha": "dc94b6ee04a05ddb9f7ae050712ff30a13149164", diff --git a/modules/nf-core/bwa/index/environment.yml b/modules/nf-core/bwa/index/environment.yml new file mode 100644 index 0000000..d8789a2 --- /dev/null +++ b/modules/nf-core/bwa/index/environment.yml @@ -0,0 +1,5 @@ +channels: + - conda-forge + - bioconda +dependencies: + - bioconda::bwa=0.7.18 diff --git a/modules/nf-core/bwa/index/main.nf b/modules/nf-core/bwa/index/main.nf new file mode 100644 index 0000000..29d9957 --- /dev/null +++ b/modules/nf-core/bwa/index/main.nf @@ -0,0 +1,53 @@ +process BWA_INDEX { + tag "$fasta" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/bwa:0.7.18--he4a0461_0' : + 'biocontainers/bwa:0.7.18--he4a0461_0' }" + + input: + tuple val(meta), path(fasta) + + output: + tuple val(meta), path("bwa") , emit: index + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def prefix = task.ext.prefix ?: "${fasta.baseName}" + def args = task.ext.args ?: '' + """ + mkdir bwa + bwa \\ + index \\ + $args \\ + -p bwa/${prefix} \\ + $fasta + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bwa: \$(echo \$(bwa 2>&1) | sed 's/^.*Version: //; s/Contact:.*\$//') + END_VERSIONS + """ + + stub: + def prefix = task.ext.prefix ?: "${fasta.baseName}" + """ + mkdir bwa + + touch bwa/${prefix}.amb + touch bwa/${prefix}.ann + touch bwa/${prefix}.bwt + touch bwa/${prefix}.pac + touch bwa/${prefix}.sa + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bwa: \$(echo \$(bwa 2>&1) | sed 's/^.*Version: //; s/Contact:.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/bwa/index/meta.yml b/modules/nf-core/bwa/index/meta.yml new file mode 100644 index 0000000..ce5cb8f --- /dev/null +++ b/modules/nf-core/bwa/index/meta.yml @@ -0,0 +1,52 @@ +name: bwa_index +description: Create BWA index for reference genome +keywords: + - index + - fasta + - genome + - reference +tools: + - bwa: + description: | + BWA is a software package for mapping DNA sequences against + a large reference genome, such as the human genome. + homepage: http://bio-bwa.sourceforge.net/ + documentation: https://bio-bwa.sourceforge.net/bwa.shtml + arxiv: arXiv:1303.3997 + licence: ["GPL-3.0-or-later"] + identifier: "" +input: + - - meta: + type: map + description: | + Groovy Map containing reference information. + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: Input genome fasta file +output: + - index: + - meta: + type: map + description: | + Groovy Map containing reference information. + e.g. [ id:'test', single_end:false ] + pattern: "*.{amb,ann,bwt,pac,sa}" + - bwa: + type: map + description: | + Groovy Map containing reference information. + e.g. [ id:'test', single_end:false ] + pattern: "*.{amb,ann,bwt,pac,sa}" + - versions: + - versions.yml: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@drpatelh" + - "@maxulysse" +maintainers: + - "@drpatelh" + - "@maxulysse" + - "@gallvp" diff --git a/modules/nf-core/bwa/index/tests/main.nf.test b/modules/nf-core/bwa/index/tests/main.nf.test new file mode 100644 index 0000000..af33e73 --- /dev/null +++ b/modules/nf-core/bwa/index/tests/main.nf.test @@ -0,0 +1,33 @@ +nextflow_process { + + name "Test Process BWA_INDEX" + tag "modules_nfcore" + tag "modules" + tag "bwa" + tag "bwa/index" + script "../main.nf" + process "BWA_INDEX" + + test("BWA index") { + + when { + process { + """ + input[0] = [ + [id: 'test'], + file(params.modules_testdata_base_path + 'genomics/sarscov2/genome/genome.fasta', checkIfExists: true) + ] + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot(process.out).match() } + ) + } + + } + +} diff --git a/modules/nf-core/bwa/index/tests/main.nf.test.snap b/modules/nf-core/bwa/index/tests/main.nf.test.snap new file mode 100644 index 0000000..7c8f046 --- /dev/null +++ b/modules/nf-core/bwa/index/tests/main.nf.test.snap @@ -0,0 +1,47 @@ +{ + "BWA index": { + "content": [ + { + "0": [ + [ + { + "id": "test" + }, + [ + "genome.amb:md5,3a68b8b2287e07dd3f5f95f4344ba76e", + "genome.ann:md5,c32e11f6c859f166c7525a9c1d583567", + "genome.bwt:md5,0469c30a1e239dd08f68afe66fde99da", + "genome.pac:md5,983e3d2cd6f36e2546e6d25a0da78d66", + "genome.sa:md5,ab3952cabf026b48cd3eb5bccbb636d1" + ] + ] + ], + "1": [ + "versions.yml:md5,a64462ac7dfb21f4ade9b02e7f65c5bb" + ], + "index": [ + [ + { + "id": "test" + }, + [ + "genome.amb:md5,3a68b8b2287e07dd3f5f95f4344ba76e", + "genome.ann:md5,c32e11f6c859f166c7525a9c1d583567", + "genome.bwt:md5,0469c30a1e239dd08f68afe66fde99da", + "genome.pac:md5,983e3d2cd6f36e2546e6d25a0da78d66", + "genome.sa:md5,ab3952cabf026b48cd3eb5bccbb636d1" + ] + ] + ], + "versions": [ + "versions.yml:md5,a64462ac7dfb21f4ade9b02e7f65c5bb" + ] + } + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.10.1" + }, + "timestamp": "2024-05-16T11:40:09.925307" + } +} \ No newline at end of file diff --git a/modules/nf-core/bwa/index/tests/tags.yml b/modules/nf-core/bwa/index/tests/tags.yml new file mode 100644 index 0000000..28bb483 --- /dev/null +++ b/modules/nf-core/bwa/index/tests/tags.yml @@ -0,0 +1,2 @@ +bwa/index: + - modules/nf-core/bwa/index/** From d5f2e4fc93e43d2d69cdae66e5bf2d92c78c87b1 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 28 Jan 2025 12:54:54 +0100 Subject: [PATCH 026/317] Update workflows/dmscore.nf to include bwa/index --- workflows/dmscore.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 041e503..dbedefc 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -5,6 +5,7 @@ */ include { FASTQC } from '../modules/nf-core/fastqc/main' include { MULTIQC } from '../modules/nf-core/multiqc/main' +include { BWA_INDEX } from '../modules/nf-core/bwa/index/main' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' From 82720ae780353bbf00f0482c8e646b0bb2efe8ff Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 9 Apr 2025 16:04:35 +0200 Subject: [PATCH 027/317] test submit From 30025ddfa0e21a91d7d2e79a942612c1d78af7cb Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 13 Apr 2025 21:17:35 +0200 Subject: [PATCH 028/317] Update dmscore.nf to add bwa index --- workflows/dmscore.nf | 15 +++++++++++++++ 1 file changed, 15 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index dbedefc..91964a4 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -25,6 +25,9 @@ workflow DMSCORE { ch_versions = Channel.empty() ch_multiqc_files = Channel.empty() + // Define fasta file as channel for BWA index Channel + .fromPath(params.fasta, checkIfExists: true) + .set { ch_fasta } // // MODULE: Run FastQC // @@ -89,6 +92,18 @@ workflow DMSCORE { emit:multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] + // + // MODULE: BWA Index + // + BWA_INDEX ( + ch_fasta + ) + + emit: + multiqc_report = MULTIQC.out.report.toList() + versions = ch_versions + bwa_index = BWA_INDEX.out.index + } /* From fae545c20af80e1336977f43f033bd734eec26f2 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 13 Apr 2025 21:37:15 +0200 Subject: [PATCH 029/317] Update dmscore.nf --- workflows/dmscore.nf | 7 ++++--- 1 file changed, 4 insertions(+), 3 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 91964a4..e019400 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -25,9 +25,10 @@ workflow DMSCORE { ch_versions = Channel.empty() ch_multiqc_files = Channel.empty() - // Define fasta file as channel for BWA index Channel - .fromPath(params.fasta, checkIfExists: true) - .set { ch_fasta } + // Define fasta file as channel for BWA index + Channel + .fromPath(params.fasta, checkIfExists: true) + .set { ch_fasta } // // MODULE: Run FastQC // From af2ed2f6b3c696b20d9ce502872ad829bb8b0331 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 13 Apr 2025 21:42:02 +0200 Subject: [PATCH 030/317] Update dmscore.nf --- workflows/dmscore.nf | 9 +++++---- 1 file changed, 5 insertions(+), 4 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index e019400..4302236 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -17,6 +17,11 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ +// Define fasta file as channel for BWA index + Channel + .fromPath(params.fasta, checkIfExists: true) + .set { ch_fasta } + workflow DMSCORE { take: @@ -25,10 +30,6 @@ workflow DMSCORE { ch_versions = Channel.empty() ch_multiqc_files = Channel.empty() - // Define fasta file as channel for BWA index - Channel - .fromPath(params.fasta, checkIfExists: true) - .set { ch_fasta } // // MODULE: Run FastQC // From 461b426944262109bd37953efb66870d5b94d4d3 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 13 Apr 2025 21:42:39 +0200 Subject: [PATCH 031/317] Update dmscore.nf --- workflows/dmscore.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 4302236..3339c1e 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -18,9 +18,9 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc */ // Define fasta file as channel for BWA index - Channel - .fromPath(params.fasta, checkIfExists: true) - .set { ch_fasta } +Channel + .fromPath(params.fasta, checkIfExists: true) + .set { ch_fasta } workflow DMSCORE { From 30a2f2640c11f27eae15110b8c54d48cb639d60a Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 13 Apr 2025 21:47:45 +0200 Subject: [PATCH 032/317] Update dmscore.nf --- workflows/dmscore.nf | 7 ++----- 1 file changed, 2 insertions(+), 5 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 3339c1e..9202db3 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -91,9 +91,6 @@ workflow DMSCORE { [] ) - emit:multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html - versions = ch_versions // channel: [ path(versions.yml) ] - // // MODULE: BWA Index // @@ -102,8 +99,8 @@ workflow DMSCORE { ) emit: - multiqc_report = MULTIQC.out.report.toList() - versions = ch_versions + multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html + versions = ch_versions // channel: [ path(versions.yml) ] bwa_index = BWA_INDEX.out.index } From 7c22a8b00e4a841685d9af04e18849504ab78618 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 13 Apr 2025 22:21:23 +0200 Subject: [PATCH 033/317] Update dmscore.nf --- workflows/dmscore.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 9202db3..caad1b7 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -20,6 +20,7 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc // Define fasta file as channel for BWA index Channel .fromPath(params.fasta, checkIfExists: true) + .map { fasta -> tuple( { id: "ref" }, fasta ) } .set { ch_fasta } workflow DMSCORE { From fb2b47f901d6fa135e454bb5d5cd820b0f813a57 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Sun, 13 Apr 2025 22:29:16 +0200 Subject: [PATCH 034/317] add bwa mem module --- modules.json | 5 + modules/nf-core/bwa/mem/environment.yml | 13 + modules/nf-core/bwa/mem/main.nf | 74 +++++ modules/nf-core/bwa/mem/meta.yml | 111 +++++++ modules/nf-core/bwa/mem/tests/main.nf.test | 260 +++++++++++++++++ .../nf-core/bwa/mem/tests/main.nf.test.snap | 271 ++++++++++++++++++ 6 files changed, 734 insertions(+) create mode 100644 modules/nf-core/bwa/mem/environment.yml create mode 100644 modules/nf-core/bwa/mem/main.nf create mode 100644 modules/nf-core/bwa/mem/meta.yml create mode 100644 modules/nf-core/bwa/mem/tests/main.nf.test create mode 100644 modules/nf-core/bwa/mem/tests/main.nf.test.snap diff --git a/modules.json b/modules.json index 026d225..28a511b 100644 --- a/modules.json +++ b/modules.json @@ -10,6 +10,11 @@ "git_sha": "2d20463181b1c38981a02e90d3084b5f9fa8d540", "installed_by": ["modules"] }, + "bwa/mem": { + "branch": "master", + "git_sha": "a29f18660f5e3748d44d6f716241e70c942c065d", + "installed_by": ["modules"] + }, "fastqc": { "branch": "master", "git_sha": "dc94b6ee04a05ddb9f7ae050712ff30a13149164", diff --git a/modules/nf-core/bwa/mem/environment.yml b/modules/nf-core/bwa/mem/environment.yml new file mode 100644 index 0000000..ed5448a --- /dev/null +++ b/modules/nf-core/bwa/mem/environment.yml @@ -0,0 +1,13 @@ +--- +# yaml-language-server: $schema=https://raw.githubusercontent.com/nf-core/modules/master/modules/environment-schema.json +channels: + - conda-forge + - bioconda + +dependencies: + # renovate: datasource=conda depName=bioconda/bwa + - bioconda::bwa=0.7.18 + # renovate: datasource=conda depName=bioconda/htslib + - bioconda::htslib=1.21 + # renovate: datasource=conda depName=bioconda/samtools + - bioconda::samtools=1.21 diff --git a/modules/nf-core/bwa/mem/main.nf b/modules/nf-core/bwa/mem/main.nf new file mode 100644 index 0000000..3c54417 --- /dev/null +++ b/modules/nf-core/bwa/mem/main.nf @@ -0,0 +1,74 @@ +process BWA_MEM { + tag "$meta.id" + label 'process_high' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://community-cr-prod.seqera.io/docker/registry/v2/blobs/sha256/bf/bf7890f8d4e38a7586581cb7fa13401b7af1582f21d94eef969df4cea852b6da/data' : + 'community.wave.seqera.io/library/bwa_htslib_samtools:56c9f8d5201889a4' }" + + input: + tuple val(meta) , path(reads) + tuple val(meta2), path(index) + tuple val(meta3), path(fasta) + val sort_bam + + output: + tuple val(meta), path("*.bam") , emit: bam, optional: true + tuple val(meta), path("*.cram") , emit: cram, optional: true + tuple val(meta), path("*.csi") , emit: csi, optional: true + tuple val(meta), path("*.crai") , emit: crai, optional: true + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def args2 = task.ext.args2 ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def samtools_command = sort_bam ? 'sort' : 'view' + def extension = args2.contains("--output-fmt sam") ? "sam" : + args2.contains("--output-fmt cram") ? "cram": + sort_bam && args2.contains("-O cram")? "cram": + !sort_bam && args2.contains("-C") ? "cram": + "bam" + def reference = fasta && extension=="cram" ? "--reference ${fasta}" : "" + if (!fasta && extension=="cram") error "Fasta reference is required for CRAM output" + """ + INDEX=`find -L ./ -name "*.amb" | sed 's/\\.amb\$//'` + + bwa mem \\ + $args \\ + -t $task.cpus \\ + \$INDEX \\ + $reads \\ + | samtools $samtools_command $args2 ${reference} --threads $task.cpus -o ${prefix}.${extension} - + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bwa: \$(echo \$(bwa 2>&1) | sed 's/^.*Version: //; s/Contact:.*\$//') + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ + + stub: + def args2 = task.ext.args2 ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + def extension = args2.contains("--output-fmt sam") ? "sam" : + args2.contains("--output-fmt cram") ? "cram": + sort_bam && args2.contains("-O cram")? "cram": + !sort_bam && args2.contains("-C") ? "cram": + "bam" + """ + touch ${prefix}.${extension} + touch ${prefix}.csi + touch ${prefix}.crai + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bwa: \$(echo \$(bwa 2>&1) | sed 's/^.*Version: //; s/Contact:.*\$//') + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/bwa/mem/meta.yml b/modules/nf-core/bwa/mem/meta.yml new file mode 100644 index 0000000..b6f696c --- /dev/null +++ b/modules/nf-core/bwa/mem/meta.yml @@ -0,0 +1,111 @@ +name: bwa_mem +description: Performs fastq alignment to a fasta reference using BWA +keywords: + - mem + - bwa + - alignment + - map + - fastq + - bam + - sam +tools: + - bwa: + description: | + BWA is a software package for mapping DNA sequences against + a large reference genome, such as the human genome. + homepage: http://bio-bwa.sourceforge.net/ + documentation: https://bio-bwa.sourceforge.net/bwa.shtml + arxiv: arXiv:1303.3997 + licence: ["GPL-3.0-or-later"] + identifier: "biotools:bwa" +input: + - - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - reads: + type: file + description: | + List of input FastQ files of size 1 and 2 for single-end and paired-end data, + respectively. + ontologies: + - edam: "http://edamontology.org/data_2044" # Sequence + - edam: "http://edamontology.org/format_1930" # FASTQ + - - meta2: + type: map + description: | + Groovy Map containing reference information. + e.g. [ id:'test', single_end:false ] + - index: + type: file + description: BWA genome index files + pattern: "Directory containing BWA index *.{amb,ann,bwt,pac,sa}" + ontologies: + - edam: "http://edamontology.org/data_3210" # Genome index + - - meta3: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: Reference genome in FASTA format + pattern: "*.{fasta,fa}" + ontologies: + - edam: "http://edamontology.org/data_2044" # Sequence + - edam: "http://edamontology.org/format_1929" # FASTA + - - sort_bam: + type: boolean + description: use samtools sort (true) or samtools view (false) + pattern: "true or false" +output: + - bam: + - meta: + type: file + description: Output BAM file containing read alignments + - "*.bam": + type: file + description: Output BAM file containing read alignments + pattern: "*.{bam}" + ontologies: + - edam: "http://edamontology.org/format_2572" # BAM + - cram: + - meta: + type: file + description: Output CRAM file containing read alignments + - "*.cram": + type: file + description: Output CRAM file containing read alignments + pattern: "*.{cram}" + ontologies: + - edam: "http://edamontology.org/format_3462" # CRAM + - csi: + - meta: + type: file + description: Optional index file for BAM file + - "*.csi": + type: file + description: Optional index file for BAM file + pattern: "*.{csi}" + - crai: + - meta: + type: file + description: Optional index file for CRAM file + - "*.crai": + type: file + description: Optional index file for CRAM file + pattern: "*.{crai}" + - versions: + - versions.yml: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@drpatelh" + - "@jeremy1805" + - "@matthdsm" +maintainers: + - "@drpatelh" + - "@jeremy1805" + - "@matthdsm" diff --git a/modules/nf-core/bwa/mem/tests/main.nf.test b/modules/nf-core/bwa/mem/tests/main.nf.test new file mode 100644 index 0000000..5de2c2f --- /dev/null +++ b/modules/nf-core/bwa/mem/tests/main.nf.test @@ -0,0 +1,260 @@ +nextflow_process { + + name "Test Process BWA_MEM" + tag "modules_nfcore" + tag "modules" + tag "bwa" + tag "bwa/mem" + tag "bwa/index" + script "../main.nf" + process "BWA_MEM" + + setup { + run("BWA_INDEX") { + script "../../index/main.nf" + process { + """ + input[0] = [ + [id: 'test'], + file(params.modules_testdata_base_path + 'genomics/sarscov2/genome/genome.fasta', checkIfExists: true) + ] + """ + } + } + } + + test("Single-End") { + + when { + process { + """ + input[0] = [ + [ id:'test', single_end:true ], // meta map + [ + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true) + ] + ] + input[1] = BWA_INDEX.out.index + input[2] = [[id: 'test'],file(params.modules_testdata_base_path + 'genomics/sarscov2/genome/genome.fasta', checkIfExists: true)] + input[3] = false + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot( + process.out.cram, + process.out.csi, + process.out.crai, + process.out.versions, + bam(process.out.bam[0][1]).getHeaderMD5(), + bam(process.out.bam[0][1]).getReadsMD5() + ).match() + } + ) + } + + } + + test("Single-End Sort") { + + when { + process { + """ + input[0] = [ + [ id:'test', single_end:true ], // meta map + [ + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true) + ] + ] + input[1] = BWA_INDEX.out.index + input[2] = [[id: 'test'],file(params.modules_testdata_base_path + 'genomics/sarscov2/genome/genome.fasta', checkIfExists: true)] + input[3] = true + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot( + process.out.cram, + process.out.csi, + process.out.crai, + process.out.versions, + bam(process.out.bam[0][1]).getHeaderMD5(), + bam(process.out.bam[0][1]).getReadsMD5() + ).match() + } + ) + } + + } + + test("Paired-End") { + + when { + process { + """ + input[0] = [ + [ id:'test', single_end:false ], // meta map + [ + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true), + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_2.fastq.gz', checkIfExists: true) + ] + ] + input[1] = BWA_INDEX.out.index + input[2] = [[id: 'test'],file(params.modules_testdata_base_path + 'genomics/sarscov2/genome/genome.fasta', checkIfExists: true)] + input[3] = false + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot( + process.out.cram, + process.out.csi, + process.out.crai, + process.out.versions, + bam(process.out.bam[0][1]).getHeaderMD5(), + bam(process.out.bam[0][1]).getReadsMD5() + ).match() + } + ) + } + + } + + test("Paired-End Sort") { + + when { + process { + """ + input[0] = [ + [ id:'test', single_end:false ], // meta map + [ + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true), + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_2.fastq.gz', checkIfExists: true) + ] + ] + input[1] = BWA_INDEX.out.index + input[2] = [[id: 'test'],file(params.modules_testdata_base_path + 'genomics/sarscov2/genome/genome.fasta', checkIfExists: true)] + input[3] = true + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot( + process.out.cram, + process.out.csi, + process.out.crai, + process.out.versions, + bam(process.out.bam[0][1]).getHeaderMD5(), + bam(process.out.bam[0][1]).getReadsMD5() + ).match() + } + ) + } + + } + + test("Paired-End - no fasta") { + + when { + process { + """ + input[0] = [ + [ id:'test', single_end:false ], // meta map + [ + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true), + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_2.fastq.gz', checkIfExists: true) + ] + ] + input[1] = BWA_INDEX.out.index + input[2] = [[:],[]] + input[3] = false + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot( + process.out.cram, + process.out.csi, + process.out.crai, + process.out.versions, + bam(process.out.bam[0][1]).getHeaderMD5(), + bam(process.out.bam[0][1]).getReadsMD5() + ).match() + } + ) + } + + } + + test("Single-end - stub") { + + options "-stub" + + when { + process { + """ + input[0] = [ + [ id:'test', single_end:true ], // meta map + [ + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true) + ] + ] + input[1] = BWA_INDEX.out.index + input[2] = [[id: 'test'],file(params.modules_testdata_base_path + 'genomics/sarscov2/genome/genome.fasta', checkIfExists: true)] + input[3] = false + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot(process.out).match() } + ) + } + } + + test("Paired-end - stub") { + + options "-stub" + + when { + process { + """ + input[0] = [ + [ id:'test', single_end:false ], // meta map + [ + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_1.fastq.gz', checkIfExists: true), + file(params.modules_testdata_base_path + 'genomics/sarscov2/illumina/fastq/test_2.fastq.gz', checkIfExists: true) + ] + ] + input[1] = BWA_INDEX.out.index + input[2] = [[id: 'test'],file(params.modules_testdata_base_path + 'genomics/sarscov2/genome/genome.fasta', checkIfExists: true)] + input[3] = false + """ + } + } + + then { + assertAll( + { assert process.success }, + { assert snapshot(process.out).match() } + ) + } + } +} diff --git a/modules/nf-core/bwa/mem/tests/main.nf.test.snap b/modules/nf-core/bwa/mem/tests/main.nf.test.snap new file mode 100644 index 0000000..3aaefdd --- /dev/null +++ b/modules/nf-core/bwa/mem/tests/main.nf.test.snap @@ -0,0 +1,271 @@ +{ + "Single-End": { + "content": [ + [ + + ], + [ + + ], + [ + + ], + [ + "versions.yml:md5,c60680eba0f00e791c0d5a0a6e9d665f" + ], + "53df0e7b72f1f85fb28af5fec435246", + "798439cbd7fd81cbcc5078022dc5479d" + ], + "meta": { + "nf-test": "0.9.2", + "nextflow": "24.10.5" + }, + "timestamp": "2025-03-27T08:36:00.831642964" + }, + "Single-End Sort": { + "content": [ + [ + + ], + [ + + ], + [ + + ], + [ + "versions.yml:md5,c60680eba0f00e791c0d5a0a6e9d665f" + ], + "5eca502b75fefc26e8000908bf0bb3a3", + "94fcf617f5b994584c4e8d4044e16b4f" + ], + "meta": { + "nf-test": "0.9.2", + "nextflow": "24.10.5" + }, + "timestamp": "2025-03-27T08:36:16.025706238" + }, + "Paired-End": { + "content": [ + [ + + ], + [ + + ], + [ + + ], + [ + "versions.yml:md5,c60680eba0f00e791c0d5a0a6e9d665f" + ], + "fec2aafbba4637767bc4e202c71aee58", + "57aeef88ed701a8ebc8e2f0a381b2a6" + ], + "meta": { + "nf-test": "0.9.2", + "nextflow": "24.10.5" + }, + "timestamp": "2025-03-27T08:36:27.309924644" + }, + "Paired-End Sort": { + "content": [ + [ + + ], + [ + + ], + [ + + ], + [ + "versions.yml:md5,c60680eba0f00e791c0d5a0a6e9d665f" + ], + "d5ad8844218280969c1f9349bd62d057", + "af8628d9df18b2d3d4f6fd47ef2bb872" + ], + "meta": { + "nf-test": "0.9.2", + "nextflow": "24.10.5" + }, + "timestamp": "2025-03-27T08:36:45.448624985" + }, + "Single-end - stub": { + "content": [ + { + "0": [ + [ + { + "id": "test", + "single_end": true + }, + "test.bam:md5,d41d8cd98f00b204e9800998ecf8427e" + ] + ], + "1": [ + + ], + "2": [ + [ + { + "id": "test", + "single_end": true + }, + "test.csi:md5,d41d8cd98f00b204e9800998ecf8427e" + ] + ], + "3": [ + [ + { + "id": "test", + "single_end": true + }, + "test.crai:md5,d41d8cd98f00b204e9800998ecf8427e" + ] + ], + "4": [ + "versions.yml:md5,c60680eba0f00e791c0d5a0a6e9d665f" + ], + "bam": [ + [ + { + "id": "test", + "single_end": true + }, + "test.bam:md5,d41d8cd98f00b204e9800998ecf8427e" + ] + ], + "crai": [ + [ + { + "id": "test", + "single_end": true + }, + "test.crai:md5,d41d8cd98f00b204e9800998ecf8427e" + ] + ], + "cram": [ + + ], + "csi": [ + [ + { + "id": "test", + "single_end": true + }, + "test.csi:md5,d41d8cd98f00b204e9800998ecf8427e" + ] + ], + "versions": [ + "versions.yml:md5,c60680eba0f00e791c0d5a0a6e9d665f" + ] + } + ], + "meta": { + "nf-test": "0.9.2", + "nextflow": "24.10.5" + }, + "timestamp": "2025-03-27T08:37:16.211123969" + }, + "Paired-End - 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ch_fasta + // [meta, reads] + FASTQC.out.fastq, + + // [meta, index files] + BWA_INDEX.out.index, + + // [meta, fasta file] + ch_fasta, + + // val: sort_bam + false ) emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] bwa_index = BWA_INDEX.out.index + aligned_bam = BWA_MEM.out.bam } From 7fb04348f60714e580ff80d717b115393af148b9 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 13 Apr 2025 22:45:10 +0200 Subject: [PATCH 037/317] Update dmscore.nf --- workflows/dmscore.nf | 5 ++++- 1 file changed, 4 insertions(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 053896b..80030c1 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -41,6 +41,9 @@ workflow DMSCORE { ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}) ch_versions = ch_versions.mix(FASTQC.out.versions.first()) + // Normalize meta information for BWA_MEM: force ID "ref" for all reads + reads_for_bwa = FASTQC.out.fastq.map { meta, reads -> tuple( [ id: "ref" ], reads ) } + // // Collate and save software versions // @@ -105,7 +108,7 @@ workflow DMSCORE { // BWA_MEM ( // [meta, reads] - FASTQC.out.fastq, + reads_for_bwa, // [meta, index files] BWA_INDEX.out.index, From 459b8ea560b4b5f1a7b35513927d1d33c9c58277 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 13 Apr 2025 23:03:22 +0200 Subject: [PATCH 038/317] Update dmscore.nf --- workflows/dmscore.nf | 28 +++++++++++----------------- 1 file changed, 11 insertions(+), 17 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 80030c1..2021c6d 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -32,6 +32,9 @@ workflow DMSCORE { ch_versions = Channel.empty() ch_multiqc_files = Channel.empty() + ch_reads = ch_samplesheet.map { row -> tuple( [ id: row.sample ], [ file(row.fastq_1), file(row.fastq_2) ] ) } // Parse reads from samplesheet (multi-sample support) + + // // MODULE: Run FastQC // @@ -41,9 +44,6 @@ workflow DMSCORE { ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}) ch_versions = ch_versions.mix(FASTQC.out.versions.first()) - // Normalize meta information for BWA_MEM: force ID "ref" for all reads - reads_for_bwa = FASTQC.out.fastq.map { meta, reads -> tuple( [ id: "ref" ], reads ) } - // // Collate and save software versions // @@ -103,21 +103,15 @@ workflow DMSCORE { ch_fasta ) - // - // MODULE: BWA Mem - // + // Broadcast index and fasta to all samples + ch_bwa_index = BWA_INDEX.out.index.broadcast() + ch_fasta_broadcast = ch_fasta.broadcast() + BWA_MEM ( - // [meta, reads] - reads_for_bwa, - - // [meta, index files] - BWA_INDEX.out.index, - - // [meta, fasta file] - ch_fasta, - - // val: sort_bam - false + ch_reads, // each sample's reads: tuple [meta, reads] + ch_bwa_index, // broadcasted index files + ch_fasta_broadcast, // broadcasted fasta file + false // sort_bam: false for now ) emit: From b278b29d50512528885b688ec400d49a81ee657a Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 13 Apr 2025 23:09:50 +0200 Subject: [PATCH 039/317] Update dmscore.nf From c027c124011c749e904af7d4ca850b5b92df0f4a Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 14:03:07 +0200 Subject: [PATCH 040/317] Update dmscore.nf (disable broadcasting) --- workflows/dmscore.nf | 9 ++++----- 1 file changed, 4 insertions(+), 5 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 2021c6d..b61e2dd 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -18,7 +18,7 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -// Define fasta file as channel for BWA index +// Define fasta file as channel (e.g. for BWA index) Channel .fromPath(params.fasta, checkIfExists: true) .map { fasta -> tuple( { id: "ref" }, fasta ) } @@ -103,14 +103,13 @@ workflow DMSCORE { ch_fasta ) - // Broadcast index and fasta to all samples - ch_bwa_index = BWA_INDEX.out.index.broadcast() - ch_fasta_broadcast = ch_fasta.broadcast() + // Broadcast index to all samples + ch_bwa_index = BWA_INDEX.out.index BWA_MEM ( ch_reads, // each sample's reads: tuple [meta, reads] ch_bwa_index, // broadcasted index files - ch_fasta_broadcast, // broadcasted fasta file + ch_fasta, // broadcasted fasta file false // sort_bam: false for now ) From 5419ded91a827732b464b239c976d7a2ebb363fe Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 14:05:16 +0200 Subject: [PATCH 041/317] Update dmscore.nf deleted ch_reads = ch_samplesheet.map { row -> tuple( [ id: row.sample ], [ file(row.fastq_1), file(row.fastq_2) ] ) } // Parse reads from samplesheet (multi-sample support) --- workflows/dmscore.nf | 2 -- 1 file changed, 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index b61e2dd..cafc0d3 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -32,8 +32,6 @@ workflow DMSCORE { ch_versions = Channel.empty() ch_multiqc_files = Channel.empty() - ch_reads = ch_samplesheet.map { row -> tuple( [ id: row.sample ], [ file(row.fastq_1), file(row.fastq_2) ] ) } // Parse reads from samplesheet (multi-sample support) - // // MODULE: Run FastQC From 7484c6e556f9b171f580e3ccf81ed1e04a0bdcfa Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 14:08:38 +0200 Subject: [PATCH 042/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index cafc0d3..7e5404c 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -105,7 +105,7 @@ workflow DMSCORE { ch_bwa_index = BWA_INDEX.out.index BWA_MEM ( - ch_reads, // each sample's reads: tuple [meta, reads] + ch_samplesheet, // each sample's reads: tuple [meta, reads] ch_bwa_index, // broadcasted index files ch_fasta, // broadcasted fasta file false // sort_bam: false for now From 381718506c70437d636bdc3e8ea717a2df0abaee Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 14:19:46 +0200 Subject: [PATCH 043/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 7e5404c..21f0120 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -108,7 +108,7 @@ workflow DMSCORE { ch_samplesheet, // each sample's reads: tuple [meta, reads] ch_bwa_index, // broadcasted index files ch_fasta, // broadcasted fasta file - false // sort_bam: false for now + false // sort_bam ) emit: From f524bd9ead1356f4f38a130151310153c7a57d92 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Tue, 15 Apr 2025 15:23:10 +0200 Subject: [PATCH 044/317] bam awk module --- modules/local/filterbamawk.nf | 90 +++++++++++++++++++++++++++++++++++ 1 file changed, 90 insertions(+) create mode 100644 modules/local/filterbamawk.nf diff --git a/modules/local/filterbamawk.nf b/modules/local/filterbamawk.nf new file mode 100644 index 0000000..195685e --- /dev/null +++ b/modules/local/filterbamawk.nf @@ -0,0 +1,90 @@ +// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) +// https://github.com/nf-core/modules/tree/master/modules/nf-core/ +// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: +// https://nf-co.re/join +// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. +// All other parameters MUST be provided using the "task.ext" directive, see here: +// https://www.nextflow.io/docs/latest/process.html#ext +// where "task.ext" is a string. +// Any parameters that need to be evaluated in the context of a particular sample +// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. +// TODO nf-core: Software that can be piped together SHOULD be added to separate module files +// unless there is a run-time, storage advantage in implementing in this way +// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: +// bwa mem | samtools view -B -T ref.fasta +// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty +// list (`[]`) instead of a file can be used to work around this issue. + +process FILTERBAMAWK { + tag '$bam' + label 'process_single' + + // TODO nf-core: List required Conda package(s). + // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). + // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. + // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/samtools:1.21--h96c455f_1': + 'biocontainers/samtools:1.21--h96c455f_1' }" + + input: + // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" + // MUST be provided as an input via a Groovy Map called "meta". + // This information may not be required in some instances e.g. indexing reference genome files: + // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf + // TODO nf-core: Where applicable please provide/convert compressed files as input/output + // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. + path bam + + output: + // TODO nf-core: Named file extensions MUST be emitted for ALL output channels + path "*.bam", emit: bam + // TODO nf-core: List additional required output channels/values here + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + + // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 + // If the software is unable to output a version number on the command-line then it can be manually specified + // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf + // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) + // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive + // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter + // using the Nextflow "task" variable e.g. "--threads $task.cpus" + // TODO nf-core: Please replace the example samtools command below with your module's command + // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) + """ + samtools view -h -F 4 -F 256 -q 30 $bam | \ +samtools view -h | \ +awk '{if(\$6 !~ /I/ && \$6 !~ /D/ && \$6 !~ /N/) print \$0}' | \ +samtools view -h | \ +awk '{for(i=1;i<=NF;i++) if(\$i ~ /^NM:i:/ && \$i != "NM:i:0") {print \$0; next}} \$1 ~ /^@/' | \ +samtools view -bS > filtered_reads.bam + +cat <<-END_VERSIONS > versions.yml +"${task.process}": + samtools: \$(samtools --version |& sed '1!d ; s/samtools //') +END_VERSIONS + """ + + stub: + def args = task.ext.args ?: '' + + // TODO nf-core: A stub section should mimic the execution of the original module as best as possible + // Have a look at the following examples: + // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 + // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 + """ + touch ${prefix}.bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + filterbamawk: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ +} From 6927ee6a16d83bb6d39f28ffae4685212eba7bd8 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Tue, 15 Apr 2025 15:30:00 +0200 Subject: [PATCH 045/317] bam awk module --- .../{filterbamawk.nf => bamfilteringdms.nf} | 35 ++++++++++--------- 1 file changed, 18 insertions(+), 17 deletions(-) rename modules/local/{filterbamawk.nf => bamfilteringdms.nf} (88%) diff --git a/modules/local/filterbamawk.nf b/modules/local/bamfilteringdms.nf similarity index 88% rename from modules/local/filterbamawk.nf rename to modules/local/bamfilteringdms.nf index 195685e..579cdaa 100644 --- a/modules/local/filterbamawk.nf +++ b/modules/local/bamfilteringdms.nf @@ -15,8 +15,8 @@ // TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty // list (`[]`) instead of a file can be used to work around this issue. -process FILTERBAMAWK { - tag '$bam' +process BAMFILTERINGDMS { + tag "$meta.id" label 'process_single' // TODO nf-core: List required Conda package(s). @@ -35,11 +35,11 @@ process FILTERBAMAWK { // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf // TODO nf-core: Where applicable please provide/convert compressed files as input/output // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. - path bam + tuple val(meta), path(bam) output: // TODO nf-core: Named file extensions MUST be emitted for ALL output channels - path "*.bam", emit: bam + tuple val(meta), path("*.bam"), emit: bam // TODO nf-core: List additional required output channels/values here path "versions.yml" , emit: versions @@ -48,7 +48,7 @@ process FILTERBAMAWK { script: def args = task.ext.args ?: '' - + def prefix = task.ext.prefix ?: "${meta.id}" // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 // If the software is unable to output a version number on the command-line then it can be manually specified // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf @@ -59,22 +59,23 @@ process FILTERBAMAWK { // TODO nf-core: Please replace the example samtools command below with your module's command // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) """ - samtools view -h -F 4 -F 256 -q 30 $bam | \ -samtools view -h | \ -awk '{if(\$6 !~ /I/ && \$6 !~ /D/ && \$6 !~ /N/) print \$0}' | \ -samtools view -h | \ -awk '{for(i=1;i<=NF;i++) if(\$i ~ /^NM:i:/ && \$i != "NM:i:0") {print \$0; next}} \$1 ~ /^@/' | \ -samtools view -bS > filtered_reads.bam + samtools \\ + sort \\ + $args \\ + -@ $task.cpus \\ + -o ${prefix}.bam \\ + -T $prefix \\ + $bam -cat <<-END_VERSIONS > versions.yml -"${task.process}": - samtools: \$(samtools --version |& sed '1!d ; s/samtools //') -END_VERSIONS + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bamfilteringdms: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS """ stub: def args = task.ext.args ?: '' - + def prefix = task.ext.prefix ?: "${meta.id}" // TODO nf-core: A stub section should mimic the execution of the original module as best as possible // Have a look at the following examples: // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 @@ -84,7 +85,7 @@ END_VERSIONS cat <<-END_VERSIONS > versions.yml "${task.process}": - filterbamawk: \$(samtools --version |& sed '1!d ; s/samtools //') + bamfilteringdms: \$(samtools --version |& sed '1!d ; s/samtools //') END_VERSIONS """ } From cc67c2e52cf3c9017b7ed474ff9592eee629d885 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 15:35:22 +0200 Subject: [PATCH 046/317] Update bamfilteringdms.nf --- modules/local/bamfilteringdms.nf | 15 +++++++-------- 1 file changed, 7 insertions(+), 8 deletions(-) diff --git a/modules/local/bamfilteringdms.nf b/modules/local/bamfilteringdms.nf index 579cdaa..e48189b 100644 --- a/modules/local/bamfilteringdms.nf +++ b/modules/local/bamfilteringdms.nf @@ -59,17 +59,16 @@ process BAMFILTERINGDMS { // TODO nf-core: Please replace the example samtools command below with your module's command // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) """ - samtools \\ - sort \\ - $args \\ - -@ $task.cpus \\ - -o ${prefix}.bam \\ - -T $prefix \\ - $bam + samtools view -h -F 4 -F 256 -q 30 $bam | \ + samtools view -h | \ + awk '{if(\$6 !~ /I/ && \$6 !~ /D/ && \$6 !~ /N/) print \$0}' | \ + samtools view -h | \ + awk '{for(i=1;i<=NF;i++) if(\$i ~ /^NM:i:/ && \$i != "NM:i:0") {print \$0; next}} \$1 ~ /^@/' | \ + samtools view -bS > filtered_reads.bam cat <<-END_VERSIONS > versions.yml "${task.process}": - bamfilteringdms: \$(samtools --version |& sed '1!d ; s/samtools //') + samtools: \$(samtools --version |& sed '1!d ; s/samtools //') END_VERSIONS """ From f9878e9a68275bdc5b7103b436a5deefca8a11c7 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 15:40:55 +0200 Subject: [PATCH 047/317] Update dmscore.nf (trying to implement awk filter function) --- workflows/dmscore.nf | 6 ++++++ 1 file changed, 6 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 21f0120..8e00abe 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -7,6 +7,7 @@ include { FASTQC } from '../modules/nf-core/fastqc/main' include { MULTIQC } from '../modules/nf-core/multiqc/main' include { BWA_INDEX } from '../modules/nf-core/bwa/index/main' include { BWA_MEM } from '../modules/nf-core/bwa/mem/main' +include { BAM_FILTER } from '../modules/local/bamfilteringdms' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -111,11 +112,16 @@ workflow DMSCORE { false // sort_bam ) + BAM_FILTER ( + BWA_MEM.out.bam + ) + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] bwa_index = BWA_INDEX.out.index aligned_bam = BWA_MEM.out.bam + filtered_bam = BAM_FILTER.out.bam } From 819d102b544640816066939d87cfa8657dad3715 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 15:42:12 +0200 Subject: [PATCH 048/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 8e00abe..38abab2 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -7,7 +7,7 @@ include { FASTQC } from '../modules/nf-core/fastqc/main' include { MULTIQC } from '../modules/nf-core/multiqc/main' include { BWA_INDEX } from '../modules/nf-core/bwa/index/main' include { BWA_MEM } from '../modules/nf-core/bwa/mem/main' -include { BAM_FILTER } from '../modules/local/bamfilteringdms' +include { BAMFILTERINGDMS } from '../modules/local/bamfilteringdms' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' From b1a3fec850d3c9d346f628d8ec13d91baf025b8e Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 15:42:56 +0200 Subject: [PATCH 049/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 38abab2..1a18a26 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -112,7 +112,7 @@ workflow DMSCORE { false // sort_bam ) - BAM_FILTER ( + BAMFILTERINGDMS ( BWA_MEM.out.bam ) From 36a0f1a2c03056a38dfa6257631ba114ae696b67 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 15:43:33 +0200 Subject: [PATCH 050/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 1a18a26..f01e313 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -121,7 +121,7 @@ workflow DMSCORE { versions = ch_versions // channel: [ path(versions.yml) ] bwa_index = BWA_INDEX.out.index aligned_bam = BWA_MEM.out.bam - filtered_bam = BAM_FILTER.out.bam + filtered_bam = BAMFILTERINGDMS.out.bam } From dec4eddc83600245d42aff1a18d7223aaf9131f2 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 16:14:28 +0200 Subject: [PATCH 051/317] Update bamfilteringdms.nf --- modules/local/bamfilteringdms.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/bamfilteringdms.nf b/modules/local/bamfilteringdms.nf index e48189b..22be831 100644 --- a/modules/local/bamfilteringdms.nf +++ b/modules/local/bamfilteringdms.nf @@ -15,7 +15,7 @@ // TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty // list (`[]`) instead of a file can be used to work around this issue. -process BAMFILTERINGDMS { +process BAMFILTER_DMS { tag "$meta.id" label 'process_single' From 2c8b653d721be6c64dacfe18cbd9e5323d03dadb Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 15 Apr 2025 16:15:08 +0200 Subject: [PATCH 052/317] Update dmscore.nf --- workflows/dmscore.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index f01e313..fa757d3 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -7,7 +7,7 @@ include { FASTQC } from '../modules/nf-core/fastqc/main' include { MULTIQC } from '../modules/nf-core/multiqc/main' include { BWA_INDEX } from '../modules/nf-core/bwa/index/main' include { BWA_MEM } from '../modules/nf-core/bwa/mem/main' -include { BAMFILTERINGDMS } from '../modules/local/bamfilteringdms' +include { BAMFILTER_DMS } from '../modules/local/bamfilteringdms' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -112,7 +112,7 @@ workflow DMSCORE { false // sort_bam ) - BAMFILTERINGDMS ( + BAMFILTER_DMS ( BWA_MEM.out.bam ) @@ -121,7 +121,7 @@ workflow DMSCORE { versions = ch_versions // channel: [ path(versions.yml) ] bwa_index = BWA_INDEX.out.index aligned_bam = BWA_MEM.out.bam - filtered_bam = BAMFILTERINGDMS.out.bam + filtered_bam = BAMFILTER_DMS.out.bam } From 035252aa2ab3aba0214d3ae66d43e655eba53de9 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Sun, 27 Apr 2025 23:29:52 +0200 Subject: [PATCH 053/317] Update premerge module to output merged_reads.fastq --- modules/local/premerge.nf | 87 +++++++++++++++++++++++++++++++++++++++ 1 file changed, 87 insertions(+) create mode 100644 modules/local/premerge.nf diff --git a/modules/local/premerge.nf b/modules/local/premerge.nf new file mode 100644 index 0000000..fdf63f0 --- /dev/null +++ b/modules/local/premerge.nf @@ -0,0 +1,87 @@ +// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) +// https://github.com/nf-core/modules/tree/master/modules/nf-core/ +// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: +// https://nf-co.re/join +// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. +// All other parameters MUST be provided using the "task.ext" directive, see here: +// https://www.nextflow.io/docs/latest/process.html#ext +// where "task.ext" is a string. +// Any parameters that need to be evaluated in the context of a particular sample +// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. +// TODO nf-core: Software that can be piped together SHOULD be added to separate module files +// unless there is a run-time, storage advantage in implementing in this way +// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: +// bwa mem | samtools view -B -T ref.fasta +// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty +// list (`[]`) instead of a file can be used to work around this issue. + +process PREMERGE { + tag "$meta.id" + label 'process_medium' + + // TODO nf-core: List required Conda package(s). + // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). + // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. + // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. + conda "${moduleDir}/environment.yml" + container "community.wave.seqera.io/library/samtools_vsearch:8c029891fb10363b" + // samtools & vsearch in container + + input: + // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" + // MUST be provided as an input via a Groovy Map called "meta". + // This information may not be required in some instances e.g. indexing reference genome files: + // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf + // TODO nf-core: Where applicable please provide/convert compressed files as input/output + // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. + tuple val(meta), path(bam) + + output: + tuple val(meta), path("merged_reads.fastq"), emit: merged_reads + path "versions.yml", emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 + // If the software is unable to output a version number on the command-line then it can be manually specified + // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf + // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) + // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive + // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter + // using the Nextflow "task" variable e.g. "--threads $task.cpus" + // TODO nf-core: Please replace the example samtools command below with your module's command + // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) + """ + # Convert BAM to paired FASTQ files + samtools fastq -1 forward_reads.fastq -2 reverse_reads.fastq -0 /dev/null -s /dev/null -n $bam + + # Merge paired reads + vsearch --fastq_mergepairs forward_reads.fastq --reverse reverse_reads.fastq --fastqout merged_reads.fastq --fastq_minovlen 10 --fastq_allowmergestagger + + # Save version + cat <<-END_VERSIONS > versions.yml + "${task.process}": + premerge: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ + + stub: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: A stub section should mimic the execution of the original module as best as possible + // Have a look at the following examples: + // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 + // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 + """ + touch ${prefix}.bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + premerge: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ +} From 6e28741163c6d13388aaf2475527d89ed1db9c16 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 27 Apr 2025 23:38:22 +0200 Subject: [PATCH 054/317] Update dmscore.nf to establish premerge function --- workflows/dmscore.nf | 6 ++++++ 1 file changed, 6 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index fa757d3..55d717c 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -8,6 +8,7 @@ include { MULTIQC } from '../modules/nf-core/multiqc/main' include { BWA_INDEX } from '../modules/nf-core/bwa/index/main' include { BWA_MEM } from '../modules/nf-core/bwa/mem/main' include { BAMFILTER_DMS } from '../modules/local/bamfilteringdms' +include { PREMERGE } from '../modules/local/premerge' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -116,12 +117,17 @@ workflow DMSCORE { BWA_MEM.out.bam ) + PREMERGE( + BAMFILTER_DMS.out.bam + ) + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] bwa_index = BWA_INDEX.out.index aligned_bam = BWA_MEM.out.bam filtered_bam = BAMFILTER_DMS.out.bam + premerged_bam = PREMERGE.out.bam } From fb55249e4a22add655e91d4d7ccda023d7c87bf5 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 27 Apr 2025 23:47:15 +0200 Subject: [PATCH 055/317] Update premerge.nf --- modules/local/premerge.nf | 45 +++++++++++---------------------------- 1 file changed, 13 insertions(+), 32 deletions(-) diff --git a/modules/local/premerge.nf b/modules/local/premerge.nf index fdf63f0..951a5ed 100644 --- a/modules/local/premerge.nf +++ b/modules/local/premerge.nf @@ -19,42 +19,23 @@ process PREMERGE { tag "$meta.id" label 'process_medium' - // TODO nf-core: List required Conda package(s). - // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). - // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. - // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/samtools_vsearch:8c029891fb10363b" - // samtools & vsearch in container + container "community.wave.seqera.io/library/bwa_samtools_vsearch:28e8640725d3d8e9" input: - // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" - // MUST be provided as an input via a Groovy Map called "meta". - // This information may not be required in some instances e.g. indexing reference genome files: - // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf - // TODO nf-core: Where applicable please provide/convert compressed files as input/output - // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. tuple val(meta), path(bam) + path wt_seq output: - tuple val(meta), path("merged_reads.fastq"), emit: merged_reads + tuple val(meta), path("merged_reads.bam"), emit: bam + tuple val(meta), path("merged_reads.fastq"), emit: fastq path "versions.yml", emit: versions when: task.ext.when == null || task.ext.when script: - def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 - // If the software is unable to output a version number on the command-line then it can be manually specified - // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf - // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) - // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive - // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter - // using the Nextflow "task" variable e.g. "--threads $task.cpus" - // TODO nf-core: Please replace the example samtools command below with your module's command - // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) """ # Convert BAM to paired FASTQ files samtools fastq -1 forward_reads.fastq -2 reverse_reads.fastq -0 /dev/null -s /dev/null -n $bam @@ -62,26 +43,26 @@ process PREMERGE { # Merge paired reads vsearch --fastq_mergepairs forward_reads.fastq --reverse reverse_reads.fastq --fastqout merged_reads.fastq --fastq_minovlen 10 --fastq_allowmergestagger - # Save version + # Re-align merged reads + bwa index $wt_seq + bwa mem $wt_seq merged_reads.fastq | samtools view -Sb - > merged_reads.bam + + # Save version information cat <<-END_VERSIONS > versions.yml "${task.process}": premerge: \$(samtools --version |& sed '1!d ; s/samtools //') END_VERSIONS """ - + stub: - def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: A stub section should mimic the execution of the original module as best as possible - // Have a look at the following examples: - // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 - // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 """ - touch ${prefix}.bam + touch merged_reads.fastq + touch merged_reads.bam cat <<-END_VERSIONS > versions.yml "${task.process}": - premerge: \$(samtools --version |& sed '1!d ; s/samtools //') + premerge: dummy_version END_VERSIONS """ } From 64ada6163f6aee8d63433ba2a7b06e8c88c1be57 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 27 Apr 2025 23:48:47 +0200 Subject: [PATCH 056/317] Update dmscore.nf --- workflows/dmscore.nf | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 55d717c..e2c4b37 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -118,7 +118,8 @@ workflow DMSCORE { ) PREMERGE( - BAMFILTER_DMS.out.bam + BAMFILTER_DMS.out.bam, + ch_fasta ) emit: From 2493abe788db3c69fa0ac2b20cea32f92ea110f4 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Sun, 27 Apr 2025 23:52:17 +0200 Subject: [PATCH 057/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index e2c4b37..c2dda72 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -118,7 +118,7 @@ workflow DMSCORE { ) PREMERGE( - BAMFILTER_DMS.out.bam, + BAMFILTER_DMS.out, ch_fasta ) From 0a43cf775a90f0b938dc9ce5c973aa1a669d47df Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Mon, 28 Apr 2025 00:00:00 +0200 Subject: [PATCH 058/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index c2dda72..e2c4b37 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -118,7 +118,7 @@ workflow DMSCORE { ) PREMERGE( - BAMFILTER_DMS.out, + BAMFILTER_DMS.out.bam, ch_fasta ) From b42b35e55a91e575e350bf6071526efc557b4002 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Mon, 28 Apr 2025 00:09:22 +0200 Subject: [PATCH 059/317] commented dmscore.nf --- workflows/dmscore.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index e2c4b37..2669a30 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -121,6 +121,7 @@ workflow DMSCORE { BAMFILTER_DMS.out.bam, ch_fasta ) + // try to use bwa_mem module inside of premerge script. Give only ch_bwa_index as an input instead of re-creating it -> see if error resolves automatically... emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html From 9f9800cf3cdca83b41a30f484e49de336c1c4aa5 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 29 Apr 2025 23:04:13 +0200 Subject: [PATCH 060/317] Update premerge.nf --- modules/local/premerge.nf | 1 - 1 file changed, 1 deletion(-) diff --git a/modules/local/premerge.nf b/modules/local/premerge.nf index 951a5ed..b1bb305 100644 --- a/modules/local/premerge.nf +++ b/modules/local/premerge.nf @@ -28,7 +28,6 @@ process PREMERGE { output: tuple val(meta), path("merged_reads.bam"), emit: bam - tuple val(meta), path("merged_reads.fastq"), emit: fastq path "versions.yml", emit: versions when: From 8a82974a9c2591e9b6277c0fdb08bbc3ce50efda Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 29 Apr 2025 23:14:28 +0200 Subject: [PATCH 061/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 2669a30..34ddc24 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -118,7 +118,7 @@ workflow DMSCORE { ) PREMERGE( - BAMFILTER_DMS.out.bam, + BAMFILTER_DMS.out, ch_fasta ) // try to use bwa_mem module inside of premerge script. Give only ch_bwa_index as an input instead of re-creating it -> see if error resolves automatically... From 49da643947a3e0ec93f38a89af80eb17a3edc815 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 29 Apr 2025 23:16:43 +0200 Subject: [PATCH 062/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 34ddc24..2669a30 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -118,7 +118,7 @@ workflow DMSCORE { ) PREMERGE( - BAMFILTER_DMS.out, + BAMFILTER_DMS.out.bam, ch_fasta ) // try to use bwa_mem module inside of premerge script. Give only ch_bwa_index as an input instead of re-creating it -> see if error resolves automatically... From e7f26faf7cc97f38f32240929d1ab946f34df20f Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 29 Apr 2025 23:32:51 +0200 Subject: [PATCH 063/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 2669a30..0184d11 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -119,7 +119,7 @@ workflow DMSCORE { PREMERGE( BAMFILTER_DMS.out.bam, - ch_fasta + ch_fasta.map{ it[1] } // extrahiere fasta path aus dem Tuple ) // try to use bwa_mem module inside of premerge script. Give only ch_bwa_index as an input instead of re-creating it -> see if error resolves automatically... From 984f6f16ab6b03f8e8e5795e3a18ea10370b0a13 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Wed, 30 Apr 2025 00:52:54 +0200 Subject: [PATCH 064/317] add gatk --- log.txt | 217 ++++++++++++++++++++ modules/local/gatk/saturationmutagenesis.nf | 55 +++++ 2 files changed, 272 insertions(+) create mode 100644 log.txt create mode 100644 modules/local/gatk/saturationmutagenesis.nf diff --git a/log.txt b/log.txt new file mode 100644 index 0000000..416172c --- /dev/null +++ b/log.txt @@ -0,0 +1,217 @@ +Apr-29 23:09:17.753 [main] DEBUG nextflow.cli.Launcher - $> nextflow run . -profile test,docker --fasta /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta --outdir ./results +Apr-29 23:09:17.863 [main] DEBUG nextflow.cli.CmdRun - N E X T F L O W ~ version 24.04.4 +Apr-29 23:09:17.894 [main] DEBUG nextflow.plugin.PluginsFacade - Setting up plugin manager > mode=prod; embedded=false; plugins-dir=/Users/benjaminwehnert/.nextflow/plugins; core-plugins: nf-amazon@2.5.3,nf-azure@1.6.1,nf-cloudcache@0.4.1,nf-codecommit@0.2.1,nf-console@1.1.3,nf-ga4gh@1.3.0,nf-google@1.13.2-patch1,nf-tower@1.9.1,nf-wave@1.4.2-patch1 +Apr-29 23:09:17.908 [main] INFO o.pf4j.DefaultPluginStatusProvider - Enabled plugins: [] +Apr-29 23:09:17.909 [main] INFO o.pf4j.DefaultPluginStatusProvider - Disabled plugins: [] +Apr-29 23:09:17.913 [main] INFO org.pf4j.DefaultPluginManager - PF4J version 3.12.0 in 'deployment' mode +Apr-29 23:09:17.925 [main] INFO org.pf4j.AbstractPluginManager - No plugins +Apr-29 23:09:18.797 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` +Apr-29 23:09:18.814 [main] DEBUG nextflow.config.ConfigBuilder - Found config local: /Users/benjaminwehnert/dmscore/nextflow.config +Apr-29 23:09:18.815 [main] DEBUG nextflow.config.ConfigBuilder - Parsing config file: /Users/benjaminwehnert/dmscore/nextflow.config +Apr-29 23:09:18.830 [main] DEBUG n.secret.LocalSecretsProvider - Secrets store: /Users/benjaminwehnert/.nextflow/secrets/store.json +Apr-29 23:09:18.833 [main] DEBUG nextflow.secret.SecretsLoader - Discovered secrets providers: [nextflow.secret.LocalSecretsProvider@606f81b5] - activable => nextflow.secret.LocalSecretsProvider@606f81b5 +Apr-29 23:09:18.843 [main] DEBUG nextflow.config.ConfigBuilder - Applying config profile: `test,docker` +Apr-29 23:09:20.216 [main] DEBUG nextflow.config.ConfigBuilder - Available config profiles: [bih, cfc_dev, uzl_omics, ifb_core, embl_hd, denbi_qbic, alice, mjolnir_globe, uppmax, giga, incliva, ilifu, ki_luria, uge, icr_alma, rosalind_uge, lugh, mccleary, unibe_ibu, vai, czbiohub_aws, jax, roslin, ccga_med, tes, scw, unc_longleaf, tigem, tubingen_apg, google, apollo, ipop_up, vsc_calcua, pdc_kth, googlels, ceci_nic5, humantechnopole, stjude, daisybio, eddie, medair, biowulf, apptainer, bi, bigpurple, adcra, cedars, pawsey_setonix, vsc_kul_uhasselt, pawsey_nimbus, ucl_myriad, utd_ganymede, charliecloud, seattlechildrens, icr_davros, ceres, arm, munin, rosalind, hasta, cfc, uzh, shu_bmrc, ebi_codon_slurm, ebc, ccga_dx, crick, ku_sund_danhead, marvin, shifter, biohpc_gen, mana, mamba, york_viking, unc_lccc, wehi, awsbatch, wustl_htcf, arcc, ceci_dragon2, imperial, maestro, software_license, cannon, genotoul, nci_gadi, abims, janelia, nu_genomics, googlebatch, oist, sahmri, kaust, alliance_canada, mpcdf, leicester, vsc_ugent, create, sage, cambridge, jex, podman, ebi_codon, cheaha, xanadu, nyu_hpc, test, marjorie, computerome, ucd_sonic, seg_globe, sanger, dkfz, bluebear, pasteur, einstein, ethz_euler, m3c, test_full, imb, ucl_cscluster, tuos_stanage, azurebatch, hki, seadragon, crukmi, csiro_petrichor, qmul_apocrita, wave, docker, engaging, gis, hypatia, psmn, eva, unity, cropdiversityhpc, nygc, fgcz, conda, crg, singularity, mpcdf_viper, pe2, self_hosted_runner, tufts, uw_hyak_pedslabs, binac2, debug, genouest, cbe, unsw_katana, gitpod, phoenix, seawulf, uod_hpc, fub_curta, uct_hpc, aws_tower, binac, fsu_draco] +Apr-29 23:09:20.271 [main] DEBUG nextflow.cli.CmdRun - Applied DSL=2 by global default +Apr-29 23:09:20.285 [main] DEBUG nextflow.cli.CmdRun - Launching `./main.nf` [intergalactic_northcutt] DSL2 - revision: 84101fc51c +Apr-29 23:09:20.287 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins declared=[nf-schema@2.3.0] +Apr-29 23:09:20.287 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins default=[] +Apr-29 23:09:20.287 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins resolved requirement=[nf-schema@2.3.0] +Apr-29 23:09:20.288 [main] DEBUG nextflow.plugin.PluginUpdater - Installing plugin nf-schema version: 2.3.0 +Apr-29 23:09:20.297 [main] INFO org.pf4j.AbstractPluginManager - Plugin 'nf-schema@2.3.0' resolved +Apr-29 23:09:20.298 [main] INFO org.pf4j.AbstractPluginManager - Start plugin 'nf-schema@2.3.0' +Apr-29 23:09:20.304 [main] DEBUG nextflow.plugin.BasePlugin - Plugin started nf-schema@2.3.0 +Apr-29 23:09:20.355 [main] DEBUG nextflow.Session - Session UUID: f85367ca-5bb8-4421-882b-4615ea49f7a1 +Apr-29 23:09:20.355 [main] DEBUG nextflow.Session - Run name: intergalactic_northcutt +Apr-29 23:09:20.355 [main] DEBUG nextflow.Session - Executor pool size: 8 +Apr-29 23:09:20.363 [main] DEBUG nextflow.file.FilePorter - File porter settings maxRetries=3; maxTransfers=50; pollTimeout=null +Apr-29 23:09:20.367 [main] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'FileTransfer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +Apr-29 23:09:20.390 [main] DEBUG nextflow.cli.CmdRun - + Version: 24.04.4 build 5917 + Created: 01-08-2024 07:05 UTC (09:05 CEST) + System: Mac OS X 15.0 + Runtime: Groovy 4.0.21 on OpenJDK 64-Bit Server VM 17.0.13+0 + Encoding: UTF-8 (UTF-8) + Process: 94258@MacBook-Air-von-Benjamin.local [127.0.0.1] + CPUs: 8 - Mem: 8 GB (77.5 MB) - Swap: 9 GB (1.2 GB) +Apr-29 23:09:20.402 [main] DEBUG nextflow.Session - Work-dir: /Users/benjaminwehnert/dmscore/work [Mac OS X] +Apr-29 23:09:20.403 [main] DEBUG nextflow.Session - Script base path does not exist or is not a directory: /Users/benjaminwehnert/dmscore/bin +Apr-29 23:09:20.418 [main] DEBUG nextflow.executor.ExecutorFactory - Extension executors providers=[] +Apr-29 23:09:20.426 [main] DEBUG nextflow.Session - Observer factory: DefaultObserverFactory +Apr-29 23:09:20.445 [main] DEBUG nextflow.Session - Observer factory: ValidationObserverFactory +Apr-29 23:09:20.486 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` +Apr-29 23:09:20.514 [main] DEBUG nextflow.cache.CacheFactory - Using Nextflow cache factory: nextflow.cache.DefaultCacheFactory +Apr-29 23:09:20.523 [main] DEBUG nextflow.util.CustomThreadPool - Creating default thread pool > poolSize: 9; maxThreads: 1000 +Apr-29 23:09:20.578 [main] DEBUG nextflow.Session - Session start +Apr-29 23:09:20.579 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow started -- trace file: /Users/benjaminwehnert/dmscore/results/pipeline_info/execution_trace_2025-04-29_23-09-19.txt +Apr-29 23:09:20.748 [main] DEBUG nextflow.script.ScriptRunner - > Launching execution +Apr-29 23:09:21.595 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +Apr-29 23:09:22.067 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema +Apr-29 23:09:22.068 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema +Apr-29 23:09:22.070 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +Apr-29 23:09:22.071 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema +Apr-29 23:09:22.497 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema +Apr-29 23:09:22.498 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema +Apr-29 23:09:22.500 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +Apr-29 23:09:22.501 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema +Apr-29 23:09:22.752 [main] INFO nextflow.Nextflow - +------------------------------------------------------ + ,--./,-. + ___ __ __ __ ___ /,-._.--~' + |\ | |__ __ / ` / \ |__) |__ } { + | \| | \__, \__/ | \ |___ \`-._,-`-, + `._,._,' + nf-core/dmscore 1.0.0dev +------------------------------------------------------ +Input/output options + input : https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv + outdir : ./results + +Reference genome options + genome : R64-1-1 + fasta : /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta + +Institutional config options + config_profile_name : Test profile + config_profile_description: Minimal test dataset to check pipeline function + +Generic options + trace_report_suffix : 2025-04-29_23-09-19 + +Core Nextflow options + runName : intergalactic_northcutt + containerEngine : docker + launchDir : /Users/benjaminwehnert/dmscore + workDir : /Users/benjaminwehnert/dmscore/work + projectDir : /Users/benjaminwehnert/dmscore + userName : benjaminwehnert + profile : test,docker + configFiles : /Users/benjaminwehnert/dmscore/nextflow.config + +!! Only displaying parameters that differ from the pipeline defaults !! +------------------------------------------------------ +* The nf-core framework + https://doi.org/10.1038/s41587-020-0439-x + +* Software dependencies + https://github.com/nf-core/dmscore/blob/master/CITATIONS.md + +Apr-29 23:09:22.755 [main] DEBUG n.validation.ValidationExtension - Starting parameters validation +Apr-29 23:09:23.139 [main] DEBUG nextflow.validation.SchemaEvaluator - Started validating /BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv +Apr-29 23:09:25.493 [main] DEBUG nextflow.validation.SchemaEvaluator - Validation of file 'https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv' passed! +Apr-29 23:09:26.686 [main] DEBUG n.v.FormatDirectoryPathEvaluator - Cloud blob storage paths are not supported by 'FormatDirectoryPathEvaluator': 's3://ngi-igenomes/igenomes/' +Apr-29 23:09:26.690 [main] DEBUG n.validation.ValidationExtension - Finishing parameters validation +Apr-29 23:09:26.778 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:FASTQC +Apr-29 23:09:26.781 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:FASTQC` matches process NFCORE_DMSCORE:DMSCORE:FASTQC +Apr-29 23:09:26.791 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-29 23:09:26.792 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-29 23:09:26.797 [main] DEBUG nextflow.executor.Executor - [warm up] executor > local +Apr-29 23:09:26.801 [main] DEBUG n.processor.LocalPollingMonitor - Creating local task monitor for executor 'local' > cpus=8; memory=8 GB; capacity=8; pollInterval=100ms; dumpInterval=5m +Apr-29 23:09:26.803 [main] DEBUG n.processor.TaskPollingMonitor - >>> barrier register (monitor: local) +Apr-29 23:09:26.964 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:MULTIQC +Apr-29 23:09:26.965 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:MULTIQC` matches process NFCORE_DMSCORE:DMSCORE:MULTIQC +Apr-29 23:09:26.968 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-29 23:09:26.968 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-29 23:09:26.979 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BWA_INDEX +Apr-29 23:09:26.982 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-29 23:09:26.982 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-29 23:09:26.995 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_high` matches labels `process_high` for process with name NFCORE_DMSCORE:DMSCORE:BWA_MEM +Apr-29 23:09:26.999 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-29 23:09:26.999 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-29 23:09:27.009 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS +Apr-29 23:09:27.011 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-29 23:09:27.011 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-29 23:09:27.019 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:PREMERGE +Apr-29 23:09:27.022 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-29 23:09:27.022 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-29 23:09:27.040 [main] DEBUG nextflow.Session - Config process names validation disabled as requested +Apr-29 23:09:27.046 [main] DEBUG nextflow.Session - Igniting dataflow network (17) +Apr-29 23:09:27.052 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:FASTQC +Apr-29 23:09:27.055 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:MULTIQC +Apr-29 23:09:27.055 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX +Apr-29 23:09:27.057 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_MEM +Apr-29 23:09:27.057 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS +Apr-29 23:09:27.057 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:PREMERGE +Apr-29 23:09:27.057 [main] DEBUG nextflow.script.ScriptRunner - Parsed script files: + Script_e5f965e09aa3641b: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/premerge.nf + Script_113f045ba19c2c46: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/fastqc/main.nf + Script_4f612cfd8c52e8cd: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nextflow_pipeline/main.nf + Script_215a636da7ab24a2: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfschema_plugin/main.nf + Script_68606fccdecc54d9: /Users/benjaminwehnert/dmscore/./subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf + Script_0f3077e98f6a93f6: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/mem/main.nf + Script_5c4e8d4051efa81e: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfcore_pipeline/main.nf + Script_4ff7366a79e0ef06: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/bamfilteringdms.nf + Script_5706c8cc12ed2c3e: /Users/benjaminwehnert/dmscore/./workflows/dmscore.nf + Script_c568aae5b239c1c5: /Users/benjaminwehnert/dmscore/main.nf + Script_4955288afd8ca61e: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/multiqc/main.nf + Script_25aa31c18d513c61: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/index/main.nf +Apr-29 23:09:27.058 [main] DEBUG nextflow.script.ScriptRunner - > Awaiting termination +Apr-29 23:09:27.058 [main] DEBUG nextflow.Session - Session await +Apr-29 23:09:27.157 [Actor Thread 10] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.001 s; external sort time: 0.017 s; total time: 0.018 s +Apr-29 23:09:27.157 [Actor Thread 1] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.001 s; external sort time: 0.017 s; total time: 0.018 s +Apr-29 23:09:27.162 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/52ef908653c9bf719aa469cfe21f3a30 +Apr-29 23:09:27.162 [Actor Thread 10] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/7ce3707a79436715f6c7474bc3e3e7ce +Apr-29 23:09:27.173 [Actor Thread 10] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-10246409531653428819 +Apr-29 23:09:27.173 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-17840128110603128341 +Apr-29 23:09:27.180 [Actor Thread 4] DEBUG nextflow.util.HashBuilder - [WARN] Unknown hashing type: class Script_5706c8cc12ed2c3e$_runScript_closure1$_closure4 +Apr-29 23:09:27.261 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-29 23:09:27.263 [Task submitter] INFO nextflow.Session - [60/7b7f6b] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta) +Apr-29 23:09:27.524 [FileTransfer-1] DEBUG nextflow.file.FilePorter - Copying foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq to work dir: /Users/benjaminwehnert/dmscore/work/stage-f85367ca-5bb8-4421-882b-4615ea49f7a1/ca/48d11c2e7271ee34cc948495531277/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq +Apr-29 23:09:27.524 [FileTransfer-2] DEBUG nextflow.file.FilePorter - Copying foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq to work dir: /Users/benjaminwehnert/dmscore/work/stage-f85367ca-5bb8-4421-882b-4615ea49f7a1/74/ad28840a55d8285cd0b61430cb8ed1/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq +Apr-29 23:09:29.547 [Actor Thread 1] INFO nextflow.file.FilePorter - Staging foreign file: https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq +Apr-29 23:09:31.564 [Actor Thread 1] INFO nextflow.file.FilePorter - Staging foreign file: https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq +Apr-29 23:09:32.670 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 1; name: NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/60/7b7f6b5bf9b2a8a1c4462bed3dc193] +Apr-29 23:09:32.673 [Task monitor] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'TaskFinalizer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +Apr-29 23:09:32.751 [TaskFinalizer-1] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'PublishDir' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +Apr-29 23:09:32.898 [Actor Thread 7] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process +Apr-29 23:09:39.217 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-29 23:09:39.218 [Task submitter] INFO nextflow.Session - [17/9fd3ef] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_MEM (gid1a_1_quality_1_pe) +Apr-29 23:09:43.924 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 3; name: NFCORE_DMSCORE:DMSCORE:BWA_MEM (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/17/9fd3ef8a622a9fc8a8f5c8d1b8397c] +Apr-29 23:09:43.936 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-29 23:09:43.936 [Task submitter] INFO nextflow.Session - [ca/ae6468] Submitted process > NFCORE_DMSCORE:DMSCORE:FASTQC (gid1a_1_quality_1_pe) +Apr-29 23:09:44.084 [TaskFinalizer-2] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<1:1> +Apr-29 23:09:44.084 [TaskFinalizer-2] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<2:1> +Apr-29 23:09:44.084 [TaskFinalizer-2] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<3:1> +Apr-29 23:09:44.129 [Actor Thread 4] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process +Apr-29 23:09:54.512 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 2; name: NFCORE_DMSCORE:DMSCORE:FASTQC (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/ca/ae646873842dc826f95e58e0508b6d] +Apr-29 23:09:54.536 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-29 23:09:54.537 [Task submitter] INFO nextflow.Session - [c7/952c5e] Submitted process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS (gid1a_1_quality_1_pe) +Apr-29 23:09:54.642 [Actor Thread 4] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 2; slices: 1; internal sort time: 0.003 s; external sort time: 0.001 s; total time: 0.004 s +Apr-29 23:09:54.645 [Actor Thread 4] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/1172d3993855a8b4f3a6ae35d54320f1 +Apr-29 23:09:54.649 [Actor Thread 4] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-3409379234343285724 +Apr-29 23:09:58.685 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 4; name: NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/c7/952c5eaa65f9728c2a31229c8816fd] +Apr-29 23:09:58.694 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-29 23:09:58.695 [Task submitter] INFO nextflow.Session - [ab/48ed5b] Submitted process > NFCORE_DMSCORE:DMSCORE:MULTIQC +Apr-29 23:09:58.700 [Actor Thread 13] DEBUG nextflow.processor.TaskProcessor - Handling unexpected condition for + task: name=NFCORE_DMSCORE:DMSCORE:PREMERGE (1); work-dir=null + error [nextflow.exception.ProcessUnrecoverableException]: Not a valid path value type: Script_5706c8cc12ed2c3e$_runScript_closure1$_closure4 () +Apr-29 23:09:58.727 [Actor Thread 13] ERROR nextflow.processor.TaskProcessor - Error executing process > 'NFCORE_DMSCORE:DMSCORE:PREMERGE (1)' + +Caused by: + Not a valid path value type: Script_5706c8cc12ed2c3e$_runScript_closure1$_closure4 () + + + +Tip: you can try to figure out what's wrong by changing to the process work dir and showing the script file named `.command.sh` +Apr-29 23:09:58.729 [Actor Thread 13] INFO nextflow.Session - Execution cancelled -- Finishing pending tasks before exit +Apr-29 23:09:58.743 [Actor Thread 13] ERROR nextflow.Nextflow - Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting +Apr-29 23:09:58.744 [main] DEBUG nextflow.Session - Session await > all processes finished +Apr-29 23:09:58.748 [Actor Thread 10] DEBUG nextflow.processor.TaskProcessor - Handling unexpected condition for + task: name=NFCORE_DMSCORE:DMSCORE:PREMERGE; work-dir=null + error [java.lang.InterruptedException]: java.lang.InterruptedException +Apr-29 23:10:08.729 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 5; name: NFCORE_DMSCORE:DMSCORE:MULTIQC; status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/ab/48ed5bed903c8f8bdbc4507c19909f] +Apr-29 23:10:08.739 [Task monitor] DEBUG n.processor.TaskPollingMonitor - <<< barrier arrives (monitor: local) - terminating tasks monitor poll loop +Apr-29 23:10:08.740 [main] DEBUG nextflow.Session - Session await > all barriers passed +Apr-29 23:10:08.766 [TaskFinalizer-5] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:MULTIQC > Skipping output binding because one or more optional files are missing: fileoutparam<2> +Apr-29 23:10:08.802 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'TaskFinalizer' shutdown completed (hard=false) +Apr-29 23:10:08.804 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'PublishDir' shutdown completed (hard=false) +Apr-29 23:10:08.818 [main] INFO nextflow.Nextflow - -[nf-core/dmscore] Pipeline completed with errors- +Apr-29 23:10:08.826 [main] DEBUG n.trace.WorkflowStatsObserver - Workflow completed > WorkflowStats[succeededCount=5; failedCount=0; ignoredCount=0; cachedCount=0; pendingCount=0; submittedCount=0; runningCount=0; retriesCount=0; abortedCount=0; succeedDuration=1m 1s; failedDuration=0ms; cachedDuration=0ms;loadCpus=0; loadMemory=0; peakRunning=2; peakCpus=8; peakMemory=16 GB; ] +Apr-29 23:10:08.826 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow completed -- saving trace file +Apr-29 23:10:08.835 [main] DEBUG nextflow.trace.ReportObserver - Workflow completed -- rendering execution report +Apr-29 23:10:10.258 [main] DEBUG nextflow.trace.TimelineObserver - Workflow completed -- rendering execution timeline +Apr-29 23:10:10.573 [main] DEBUG nextflow.cache.CacheDB - Closing CacheDB done +Apr-29 23:10:10.593 [main] INFO org.pf4j.AbstractPluginManager - Stop plugin 'nf-schema@2.3.0' +Apr-29 23:10:10.593 [main] DEBUG nextflow.plugin.BasePlugin - Plugin stopped nf-schema +Apr-29 23:10:10.594 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'FileTransfer' shutdown completed (hard=false) +Apr-29 23:10:10.597 [main] DEBUG nextflow.script.ScriptRunner - > Execution complete -- Goodbye diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf new file mode 100644 index 0000000..526fa3d --- /dev/null +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -0,0 +1,55 @@ +process GATK_SATURATIONMUTAGENESIS { + tag "$meta.id" + label 'process_medium' + + conda "${moduleDir}/environment.yml" + container "community.wave.seqera.io/library/gatk_samtools:YOUR_TAG_HERE" + + input: + tuple val(meta), path(premerged_reads) + path wt_seq + path pos_range + val min_counts + + output: + tuple val(meta), path("gatk_output.*"), emit: gatk_output + path "versions.yml", emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def prefix = task.ext.prefix ?: "${meta.id}" + """ + # Index reference + samtools faidx $wt_seq + gatk CreateSequenceDictionary -R $wt_seq + + # Read start and stop codon from input + start_stop_codon=\$(cat $pos_range) + + # Run GATK AnalyzeSaturationMutagenesis + gatk AnalyzeSaturationMutagenesis \ + -I $premerged_reads \ + -R $wt_seq \ + --orf \$start_stop_codon \ + --paired-mode false \ + --min-q 30 \ + --min-variant-obs $min_counts \ + -O gatk_output + + # Save versions + cat <<-END_VERSIONS > versions.yml + "${task.process}": + samtools: \$(samtools --version |& sed '1!d ; s/samtools //') + gatk: \$(gatk --version |& sed 's/^.*GATK/\1/') + END_VERSIONS + """ + + stub: + def prefix = task.ext.prefix ?: "${meta.id}" + """ + touch gatk_output.variantCounts + touch versions.yml + """ +} \ No newline at end of file From 31f3df9d8b793987a5e18505117b505cd9fdf781 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 01:10:22 +0200 Subject: [PATCH 065/317] Update dmscore.nf --- workflows/dmscore.nf | 22 ++++++++++++++++++++-- 1 file changed, 20 insertions(+), 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 0184d11..66719f9 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -26,6 +26,18 @@ Channel .map { fasta -> tuple( { id: "ref" }, fasta ) } .set { ch_fasta } +// Define reading_frame as channel (e.g. for gatk function) +Channel + .fromPath(params.reading_frame, checkIfExists: true) + .set { reading_frame_ch } + +// Define min_counts as channel (e.g. for gatk function) -> if not set - default: 3 +params.min_counts = params.min_counts ?: 3 +Channel + .value(params.min_counts) + .set { min_counts_ch } + + workflow DMSCORE { take: @@ -119,9 +131,15 @@ workflow DMSCORE { PREMERGE( BAMFILTER_DMS.out.bam, - ch_fasta.map{ it[1] } // extrahiere fasta path aus dem Tuple + ch_fasta.map{ it[1] } // extract fasta path from Tuple + ) + + GATK_SATURATIONMUTAGENESIS( + PREMERGE.out.bam, // merged reads (tuple(meta, merged_reads.fastq)) + ch_fasta.map{ it[1] }, // fasta path (path) + reading_frame_ch, // codon range (path) + min_counts_ch // min_counts (val) ) - // try to use bwa_mem module inside of premerge script. Give only ch_bwa_index as an input instead of re-creating it -> see if error resolves automatically... emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html From 86bbd665ec4207ac31343426d85a790c1eb42f97 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 01:15:11 +0200 Subject: [PATCH 066/317] Update dmscore.nf --- workflows/dmscore.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 66719f9..2545d96 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -9,6 +9,7 @@ include { BWA_INDEX } from '../modules/nf-core/bwa/index/main' include { BWA_MEM } from '../modules/nf-core/bwa/mem/main' include { BAMFILTER_DMS } from '../modules/local/bamfilteringdms' include { PREMERGE } from '../modules/local/premerge' +include { GATK_SATURATIONMUTAGENESIS } from '../modules/local/gatk/saturationmutagenesis' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' From c3a064b792003ae0bbd43252e8cb14f339363149 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 01:20:43 +0200 Subject: [PATCH 067/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index 526fa3d..257c104 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -8,7 +8,7 @@ process GATK_SATURATIONMUTAGENESIS { input: tuple val(meta), path(premerged_reads) path wt_seq - path pos_range + val pos_range val min_counts output: @@ -52,4 +52,4 @@ process GATK_SATURATIONMUTAGENESIS { touch gatk_output.variantCounts touch versions.yml """ -} \ No newline at end of file +} From d62f1a824880246cc2da3eb3c5359de6a0271d0c Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 01:21:22 +0200 Subject: [PATCH 068/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 2545d96..ea776e2 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -29,7 +29,7 @@ Channel // Define reading_frame as channel (e.g. for gatk function) Channel - .fromPath(params.reading_frame, checkIfExists: true) + .value(params.reading_frame, checkIfExists: true) .set { reading_frame_ch } // Define min_counts as channel (e.g. for gatk function) -> if not set - default: 3 From 8fdae75ac762248f0b44894b1e9f5719e452aaad Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 01:25:14 +0200 Subject: [PATCH 069/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index ea776e2..55e3fd6 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -29,7 +29,7 @@ Channel // Define reading_frame as channel (e.g. for gatk function) Channel - .value(params.reading_frame, checkIfExists: true) + .value(params.reading_frame) .set { reading_frame_ch } // Define min_counts as channel (e.g. for gatk function) -> if not set - default: 3 From 0b2ad3e8287e6132b15ed0780b5ad86e60b94828 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 01:28:08 +0200 Subject: [PATCH 070/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index 257c104..d049c67 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -26,7 +26,7 @@ process GATK_SATURATIONMUTAGENESIS { gatk CreateSequenceDictionary -R $wt_seq # Read start and stop codon from input - start_stop_codon=\$(cat $pos_range) + start_stop_codon="$pos_range" # Run GATK AnalyzeSaturationMutagenesis gatk AnalyzeSaturationMutagenesis \ From f9cfa1418551e77bec0e3fbd5e7a9d87fae0c227 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 01:31:40 +0200 Subject: [PATCH 071/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index d049c67..0e38c1a 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -3,7 +3,7 @@ process GATK_SATURATIONMUTAGENESIS { label 'process_medium' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/gatk_samtools:YOUR_TAG_HERE" + container "community.wave.seqera.io/library/gatk:3.8--6d58a01a86578bc5" input: tuple val(meta), path(premerged_reads) From 1f5d605806603937ff6038bc4bfbd1a65dc3077f Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 01:36:52 +0200 Subject: [PATCH 072/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index 0e38c1a..4bce52d 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -3,7 +3,7 @@ process GATK_SATURATIONMUTAGENESIS { label 'process_medium' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/gatk:3.8--6d58a01a86578bc5" + container "community.wave.seqera.io/library/gatk:3.8--3a1d527da3fc6d32" input: tuple val(meta), path(premerged_reads) From 2ab9b3e4f197b21efe0fbc9a87cff07d85c21535 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 01:51:28 +0200 Subject: [PATCH 073/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index 4bce52d..30e11d8 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -3,7 +3,7 @@ process GATK_SATURATIONMUTAGENESIS { label 'process_medium' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/gatk:3.8--3a1d527da3fc6d32" + container "community.wave.seqera.io/library/gatk_samtools_java-1.7.0-openjdk-conda-aarch64:50bbe1742b62bc8f" input: tuple val(meta), path(premerged_reads) From 0f86191e7afab88dcd63d94e6a40ed1244eeb0d6 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 02:13:33 +0200 Subject: [PATCH 074/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index 30e11d8..07385bd 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -3,7 +3,7 @@ process GATK_SATURATIONMUTAGENESIS { label 'process_medium' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/gatk_samtools_java-1.7.0-openjdk-conda-aarch64:50bbe1742b62bc8f" + container "community.wave.seqera.io/library/gatk4_samtools_java-1.7.0-openjdk-conda-aarch64:7c1f89018b5d5103" input: tuple val(meta), path(premerged_reads) From 8d58d28c895bf282934108b5bbe69285bb187137 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 14:14:49 +0200 Subject: [PATCH 075/317] Update nextflow_schema.json --- nextflow_schema.json | 14 +++++++++++++- 1 file changed, 13 insertions(+), 1 deletion(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 2103634..74cdacc 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -40,7 +40,19 @@ "type": "string", "description": "MultiQC report title. Printed as page header, used for filename if not otherwise specified.", "fa_icon": "fas fa-file-signature" - } + }, + "reading_frame": { + "type": "string", + "description": "Start and stop codon positions in the format 'start-stop', e.g., '352-1383'.", + "pattern": "^\\d+-\\d+$", + "default": "352-1383" + }, + "min_counts": { + "type": "integer", + "description": "Minimum number of variant observations required.", + "minimum": 1, + "default": 3 + } } }, "reference_genome_options": { From 153216848551e7614d657c4e9f8729c81f3a876c Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 30 Apr 2025 14:17:26 +0200 Subject: [PATCH 076/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index 07385bd..3af1bf5 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -1,6 +1,6 @@ process GATK_SATURATIONMUTAGENESIS { tag "$meta.id" - label 'process_medium' + label 'process_high' conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/gatk4_samtools_java-1.7.0-openjdk-conda-aarch64:7c1f89018b5d5103" From 73d41cd41bdcfe50b437fab9d6474c9f73de0fd8 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Wed, 30 Apr 2025 16:28:44 +0200 Subject: [PATCH 077/317] folder structure for R related processes --- log.txt | 375 ++++++++++++--------- modules/local/dmsanalysis/aaseq.nf | 91 +++++ modules/local/dmsanalysis/annotategatk.nf | 91 +++++ modules/local/dmsanalysis/visualization.nf | 91 +++++ 4 files changed, 487 insertions(+), 161 deletions(-) create mode 100644 modules/local/dmsanalysis/aaseq.nf create mode 100644 modules/local/dmsanalysis/annotategatk.nf create mode 100644 modules/local/dmsanalysis/visualization.nf diff --git a/log.txt b/log.txt index 416172c..59304d4 100644 --- a/log.txt +++ b/log.txt @@ -1,60 +1,61 @@ -Apr-29 23:09:17.753 [main] DEBUG nextflow.cli.Launcher - $> nextflow run . -profile test,docker --fasta /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta --outdir ./results -Apr-29 23:09:17.863 [main] DEBUG nextflow.cli.CmdRun - N E X T F L O W ~ version 24.04.4 -Apr-29 23:09:17.894 [main] DEBUG nextflow.plugin.PluginsFacade - Setting up plugin manager > mode=prod; embedded=false; plugins-dir=/Users/benjaminwehnert/.nextflow/plugins; core-plugins: nf-amazon@2.5.3,nf-azure@1.6.1,nf-cloudcache@0.4.1,nf-codecommit@0.2.1,nf-console@1.1.3,nf-ga4gh@1.3.0,nf-google@1.13.2-patch1,nf-tower@1.9.1,nf-wave@1.4.2-patch1 -Apr-29 23:09:17.908 [main] INFO o.pf4j.DefaultPluginStatusProvider - Enabled plugins: [] -Apr-29 23:09:17.909 [main] INFO o.pf4j.DefaultPluginStatusProvider - Disabled plugins: [] -Apr-29 23:09:17.913 [main] INFO org.pf4j.DefaultPluginManager - PF4J version 3.12.0 in 'deployment' mode -Apr-29 23:09:17.925 [main] INFO org.pf4j.AbstractPluginManager - No plugins -Apr-29 23:09:18.797 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` -Apr-29 23:09:18.814 [main] DEBUG nextflow.config.ConfigBuilder - Found config local: /Users/benjaminwehnert/dmscore/nextflow.config -Apr-29 23:09:18.815 [main] DEBUG nextflow.config.ConfigBuilder - Parsing config file: /Users/benjaminwehnert/dmscore/nextflow.config -Apr-29 23:09:18.830 [main] DEBUG n.secret.LocalSecretsProvider - Secrets store: /Users/benjaminwehnert/.nextflow/secrets/store.json -Apr-29 23:09:18.833 [main] DEBUG nextflow.secret.SecretsLoader - Discovered secrets providers: [nextflow.secret.LocalSecretsProvider@606f81b5] - activable => nextflow.secret.LocalSecretsProvider@606f81b5 -Apr-29 23:09:18.843 [main] DEBUG nextflow.config.ConfigBuilder - Applying config profile: `test,docker` -Apr-29 23:09:20.216 [main] DEBUG nextflow.config.ConfigBuilder - Available config profiles: [bih, cfc_dev, uzl_omics, ifb_core, embl_hd, denbi_qbic, alice, mjolnir_globe, uppmax, giga, incliva, ilifu, ki_luria, uge, icr_alma, rosalind_uge, lugh, mccleary, unibe_ibu, vai, czbiohub_aws, jax, roslin, ccga_med, tes, scw, unc_longleaf, tigem, tubingen_apg, google, apollo, ipop_up, vsc_calcua, pdc_kth, googlels, ceci_nic5, humantechnopole, stjude, daisybio, eddie, medair, biowulf, apptainer, bi, bigpurple, adcra, cedars, pawsey_setonix, vsc_kul_uhasselt, pawsey_nimbus, ucl_myriad, utd_ganymede, charliecloud, seattlechildrens, icr_davros, ceres, arm, munin, rosalind, hasta, cfc, uzh, shu_bmrc, ebi_codon_slurm, ebc, ccga_dx, crick, ku_sund_danhead, marvin, shifter, biohpc_gen, mana, mamba, york_viking, unc_lccc, wehi, awsbatch, wustl_htcf, arcc, ceci_dragon2, imperial, maestro, software_license, cannon, genotoul, nci_gadi, abims, janelia, nu_genomics, googlebatch, oist, sahmri, kaust, alliance_canada, mpcdf, leicester, vsc_ugent, create, sage, cambridge, jex, podman, ebi_codon, cheaha, xanadu, nyu_hpc, test, marjorie, computerome, ucd_sonic, seg_globe, sanger, dkfz, bluebear, pasteur, einstein, ethz_euler, m3c, test_full, imb, ucl_cscluster, tuos_stanage, azurebatch, hki, seadragon, crukmi, csiro_petrichor, qmul_apocrita, wave, docker, engaging, gis, hypatia, psmn, eva, unity, cropdiversityhpc, nygc, fgcz, conda, crg, singularity, mpcdf_viper, pe2, self_hosted_runner, tufts, uw_hyak_pedslabs, binac2, debug, genouest, cbe, unsw_katana, gitpod, phoenix, seawulf, uod_hpc, fub_curta, uct_hpc, aws_tower, binac, fsu_draco] -Apr-29 23:09:20.271 [main] DEBUG nextflow.cli.CmdRun - Applied DSL=2 by global default -Apr-29 23:09:20.285 [main] DEBUG nextflow.cli.CmdRun - Launching `./main.nf` [intergalactic_northcutt] DSL2 - revision: 84101fc51c -Apr-29 23:09:20.287 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins declared=[nf-schema@2.3.0] -Apr-29 23:09:20.287 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins default=[] -Apr-29 23:09:20.287 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins resolved requirement=[nf-schema@2.3.0] -Apr-29 23:09:20.288 [main] DEBUG nextflow.plugin.PluginUpdater - Installing plugin nf-schema version: 2.3.0 -Apr-29 23:09:20.297 [main] INFO org.pf4j.AbstractPluginManager - Plugin 'nf-schema@2.3.0' resolved -Apr-29 23:09:20.298 [main] INFO org.pf4j.AbstractPluginManager - Start plugin 'nf-schema@2.3.0' -Apr-29 23:09:20.304 [main] DEBUG nextflow.plugin.BasePlugin - Plugin started nf-schema@2.3.0 -Apr-29 23:09:20.355 [main] DEBUG nextflow.Session - Session UUID: f85367ca-5bb8-4421-882b-4615ea49f7a1 -Apr-29 23:09:20.355 [main] DEBUG nextflow.Session - Run name: intergalactic_northcutt -Apr-29 23:09:20.355 [main] DEBUG nextflow.Session - Executor pool size: 8 -Apr-29 23:09:20.363 [main] DEBUG nextflow.file.FilePorter - File porter settings maxRetries=3; maxTransfers=50; pollTimeout=null -Apr-29 23:09:20.367 [main] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'FileTransfer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false -Apr-29 23:09:20.390 [main] DEBUG nextflow.cli.CmdRun - +Apr-30 01:54:39.166 [main] DEBUG nextflow.cli.Launcher - $> nextflow run . -profile test,docker --fasta /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta --reading_frame 352-1383 --outdir ./results -resume +Apr-30 01:54:39.306 [main] DEBUG nextflow.cli.CmdRun - N E X T F L O W ~ version 24.04.4 +Apr-30 01:54:39.333 [main] DEBUG nextflow.plugin.PluginsFacade - Setting up plugin manager > mode=prod; embedded=false; plugins-dir=/Users/benjaminwehnert/.nextflow/plugins; core-plugins: nf-amazon@2.5.3,nf-azure@1.6.1,nf-cloudcache@0.4.1,nf-codecommit@0.2.1,nf-console@1.1.3,nf-ga4gh@1.3.0,nf-google@1.13.2-patch1,nf-tower@1.9.1,nf-wave@1.4.2-patch1 +Apr-30 01:54:39.345 [main] INFO o.pf4j.DefaultPluginStatusProvider - Enabled plugins: [] +Apr-30 01:54:39.346 [main] INFO o.pf4j.DefaultPluginStatusProvider - Disabled plugins: [] +Apr-30 01:54:39.349 [main] INFO org.pf4j.DefaultPluginManager - PF4J version 3.12.0 in 'deployment' mode +Apr-30 01:54:39.366 [main] INFO org.pf4j.AbstractPluginManager - No plugins +Apr-30 01:54:40.248 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` +Apr-30 01:54:40.266 [main] DEBUG nextflow.config.ConfigBuilder - Found config local: /Users/benjaminwehnert/dmscore/nextflow.config +Apr-30 01:54:40.274 [main] DEBUG nextflow.config.ConfigBuilder - Parsing config file: /Users/benjaminwehnert/dmscore/nextflow.config +Apr-30 01:54:40.290 [main] DEBUG n.secret.LocalSecretsProvider - Secrets store: /Users/benjaminwehnert/.nextflow/secrets/store.json +Apr-30 01:54:40.293 [main] DEBUG nextflow.secret.SecretsLoader - Discovered secrets providers: [nextflow.secret.LocalSecretsProvider@606f81b5] - activable => nextflow.secret.LocalSecretsProvider@606f81b5 +Apr-30 01:54:40.304 [main] DEBUG nextflow.config.ConfigBuilder - Applying config profile: `test,docker` +Apr-30 01:54:41.801 [main] DEBUG nextflow.config.ConfigBuilder - Available config profiles: [bih, cfc_dev, uzl_omics, ifb_core, embl_hd, denbi_qbic, alice, mjolnir_globe, uppmax, giga, incliva, ilifu, ki_luria, uge, icr_alma, rosalind_uge, lugh, mccleary, unibe_ibu, vai, czbiohub_aws, jax, roslin, ccga_med, tes, scw, unc_longleaf, tigem, tubingen_apg, google, apollo, ipop_up, vsc_calcua, pdc_kth, googlels, ceci_nic5, humantechnopole, stjude, daisybio, eddie, medair, biowulf, apptainer, bi, bigpurple, adcra, cedars, pawsey_setonix, vsc_kul_uhasselt, pawsey_nimbus, ucl_myriad, utd_ganymede, charliecloud, seattlechildrens, icr_davros, ceres, arm, munin, rosalind, hasta, cfc, uzh, shu_bmrc, ebi_codon_slurm, ebc, ccga_dx, crick, ku_sund_danhead, marvin, shifter, biohpc_gen, mana, mamba, york_viking, unc_lccc, wehi, awsbatch, wustl_htcf, arcc, ceci_dragon2, imperial, maestro, software_license, cannon, genotoul, nci_gadi, abims, janelia, nu_genomics, googlebatch, oist, sahmri, kaust, alliance_canada, mpcdf, leicester, vsc_ugent, create, sage, cambridge, jex, podman, ebi_codon, cheaha, xanadu, nyu_hpc, test, marjorie, computerome, ucd_sonic, seg_globe, sanger, dkfz, bluebear, pasteur, einstein, ethz_euler, m3c, test_full, imb, ucl_cscluster, tuos_stanage, azurebatch, hki, seadragon, crukmi, csiro_petrichor, qmul_apocrita, wave, docker, engaging, gis, hypatia, psmn, eva, unity, cropdiversityhpc, nygc, fgcz, conda, crg, singularity, mpcdf_viper, pe2, self_hosted_runner, tufts, uw_hyak_pedslabs, binac2, debug, genouest, cbe, unsw_katana, gitpod, phoenix, seawulf, uod_hpc, fub_curta, uct_hpc, aws_tower, binac, fsu_draco] +Apr-30 01:54:41.868 [main] DEBUG nextflow.cli.CmdRun - Applied DSL=2 by global default +Apr-30 01:54:41.893 [main] DEBUG nextflow.cli.CmdRun - Launching `./main.nf` [jolly_cori] DSL2 - revision: 84101fc51c +Apr-30 01:54:41.895 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins declared=[nf-schema@2.3.0] +Apr-30 01:54:41.895 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins default=[] +Apr-30 01:54:41.895 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins resolved requirement=[nf-schema@2.3.0] +Apr-30 01:54:41.896 [main] DEBUG nextflow.plugin.PluginUpdater - Installing plugin nf-schema version: 2.3.0 +Apr-30 01:54:41.905 [main] INFO org.pf4j.AbstractPluginManager - Plugin 'nf-schema@2.3.0' resolved +Apr-30 01:54:41.906 [main] INFO org.pf4j.AbstractPluginManager - Start plugin 'nf-schema@2.3.0' +Apr-30 01:54:41.913 [main] DEBUG nextflow.plugin.BasePlugin - Plugin started nf-schema@2.3.0 +Apr-30 01:54:41.988 [main] DEBUG nextflow.Session - Session UUID: 36458275-f468-409a-bb18-98996facae84 +Apr-30 01:54:41.989 [main] DEBUG nextflow.Session - Run name: jolly_cori +Apr-30 01:54:41.989 [main] DEBUG nextflow.Session - Executor pool size: 8 +Apr-30 01:54:41.999 [main] DEBUG nextflow.file.FilePorter - File porter settings maxRetries=3; maxTransfers=50; pollTimeout=null +Apr-30 01:54:42.005 [main] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'FileTransfer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +Apr-30 01:54:42.029 [main] DEBUG nextflow.cli.CmdRun - Version: 24.04.4 build 5917 Created: 01-08-2024 07:05 UTC (09:05 CEST) System: Mac OS X 15.0 Runtime: Groovy 4.0.21 on OpenJDK 64-Bit Server VM 17.0.13+0 Encoding: UTF-8 (UTF-8) - Process: 94258@MacBook-Air-von-Benjamin.local [127.0.0.1] - CPUs: 8 - Mem: 8 GB (77.5 MB) - Swap: 9 GB (1.2 GB) -Apr-29 23:09:20.402 [main] DEBUG nextflow.Session - Work-dir: /Users/benjaminwehnert/dmscore/work [Mac OS X] -Apr-29 23:09:20.403 [main] DEBUG nextflow.Session - Script base path does not exist or is not a directory: /Users/benjaminwehnert/dmscore/bin -Apr-29 23:09:20.418 [main] DEBUG nextflow.executor.ExecutorFactory - Extension executors providers=[] -Apr-29 23:09:20.426 [main] DEBUG nextflow.Session - Observer factory: DefaultObserverFactory -Apr-29 23:09:20.445 [main] DEBUG nextflow.Session - Observer factory: ValidationObserverFactory -Apr-29 23:09:20.486 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` -Apr-29 23:09:20.514 [main] DEBUG nextflow.cache.CacheFactory - Using Nextflow cache factory: nextflow.cache.DefaultCacheFactory -Apr-29 23:09:20.523 [main] DEBUG nextflow.util.CustomThreadPool - Creating default thread pool > poolSize: 9; maxThreads: 1000 -Apr-29 23:09:20.578 [main] DEBUG nextflow.Session - Session start -Apr-29 23:09:20.579 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow started -- trace file: /Users/benjaminwehnert/dmscore/results/pipeline_info/execution_trace_2025-04-29_23-09-19.txt -Apr-29 23:09:20.748 [main] DEBUG nextflow.script.ScriptRunner - > Launching execution -Apr-29 23:09:21.595 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema -Apr-29 23:09:22.067 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema -Apr-29 23:09:22.068 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema -Apr-29 23:09:22.070 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema -Apr-29 23:09:22.071 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema -Apr-29 23:09:22.497 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema -Apr-29 23:09:22.498 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema -Apr-29 23:09:22.500 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema -Apr-29 23:09:22.501 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema -Apr-29 23:09:22.752 [main] INFO nextflow.Nextflow - + Process: 3502@MacBook-Air-von-Benjamin.local [127.0.0.1] + CPUs: 8 - Mem: 8 GB (138.6 MB) - Swap: 8 GB (790.1 MB) +Apr-30 01:54:42.059 [main] DEBUG nextflow.Session - Work-dir: /Users/benjaminwehnert/dmscore/work [Mac OS X] +Apr-30 01:54:42.060 [main] DEBUG nextflow.Session - Script base path does not exist or is not a directory: /Users/benjaminwehnert/dmscore/bin +Apr-30 01:54:42.073 [main] DEBUG nextflow.executor.ExecutorFactory - Extension executors providers=[] +Apr-30 01:54:42.083 [main] DEBUG nextflow.Session - Observer factory: DefaultObserverFactory +Apr-30 01:54:42.105 [main] DEBUG nextflow.Session - Observer factory: ValidationObserverFactory +Apr-30 01:54:42.139 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` +Apr-30 01:54:42.164 [main] DEBUG nextflow.cache.CacheFactory - Using Nextflow cache factory: nextflow.cache.DefaultCacheFactory +Apr-30 01:54:42.172 [main] DEBUG nextflow.util.CustomThreadPool - Creating default thread pool > poolSize: 9; maxThreads: 1000 +Apr-30 01:54:42.275 [main] DEBUG nextflow.Session - Session start +Apr-30 01:54:42.276 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow started -- trace file: /Users/benjaminwehnert/dmscore/results/pipeline_info/execution_trace_2025-04-30_01-54-40.txt +Apr-30 01:54:42.437 [main] DEBUG nextflow.script.ScriptRunner - > Launching execution +Apr-30 01:54:43.415 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +Apr-30 01:54:43.844 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema +Apr-30 01:54:43.845 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema +Apr-30 01:54:43.847 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +Apr-30 01:54:43.848 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema +Apr-30 01:54:44.121 [main] WARN nextflow.script.ScriptBinding - Access to undefined parameter `min_counts` -- Initialise it to a default value eg. `params.min_counts = some_value` +Apr-30 01:54:44.245 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema +Apr-30 01:54:44.246 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema +Apr-30 01:54:44.248 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +Apr-30 01:54:44.249 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema +Apr-30 01:54:44.452 [main] INFO nextflow.Nextflow - ------------------------------------------------------ ,--./,-.  ___ __ __ __ ___ /,-._.--~' @@ -76,10 +77,10 @@ Apr-29 23:09:22.752 [main] INFO nextflow.Nextflow - config_profile_description: Minimal test dataset to check pipeline function Generic options - trace_report_suffix : 2025-04-29_23-09-19 + trace_report_suffix : 2025-04-30_01-54-40 Core Nextflow options - runName : intergalactic_northcutt + runName : jolly_cori containerEngine : docker launchDir : /Users/benjaminwehnert/dmscore workDir : /Users/benjaminwehnert/dmscore/work @@ -96,122 +97,174 @@ Apr-29 23:09:22.752 [main] INFO nextflow.Nextflow - * Software dependencies https://github.com/nf-core/dmscore/blob/master/CITATIONS.md -Apr-29 23:09:22.755 [main] DEBUG n.validation.ValidationExtension - Starting parameters validation -Apr-29 23:09:23.139 [main] DEBUG nextflow.validation.SchemaEvaluator - Started validating /BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv -Apr-29 23:09:25.493 [main] DEBUG nextflow.validation.SchemaEvaluator - Validation of file 'https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv' passed! -Apr-29 23:09:26.686 [main] DEBUG n.v.FormatDirectoryPathEvaluator - Cloud blob storage paths are not supported by 'FormatDirectoryPathEvaluator': 's3://ngi-igenomes/igenomes/' -Apr-29 23:09:26.690 [main] DEBUG n.validation.ValidationExtension - Finishing parameters validation -Apr-29 23:09:26.778 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:FASTQC -Apr-29 23:09:26.781 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:FASTQC` matches process NFCORE_DMSCORE:DMSCORE:FASTQC -Apr-29 23:09:26.791 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-29 23:09:26.792 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-29 23:09:26.797 [main] DEBUG nextflow.executor.Executor - [warm up] executor > local -Apr-29 23:09:26.801 [main] DEBUG n.processor.LocalPollingMonitor - Creating local task monitor for executor 'local' > cpus=8; memory=8 GB; capacity=8; pollInterval=100ms; dumpInterval=5m -Apr-29 23:09:26.803 [main] DEBUG n.processor.TaskPollingMonitor - >>> barrier register (monitor: local) -Apr-29 23:09:26.964 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:MULTIQC -Apr-29 23:09:26.965 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:MULTIQC` matches process NFCORE_DMSCORE:DMSCORE:MULTIQC -Apr-29 23:09:26.968 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-29 23:09:26.968 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-29 23:09:26.979 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BWA_INDEX -Apr-29 23:09:26.982 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-29 23:09:26.982 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-29 23:09:26.995 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_high` matches labels `process_high` for process with name NFCORE_DMSCORE:DMSCORE:BWA_MEM -Apr-29 23:09:26.999 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-29 23:09:26.999 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-29 23:09:27.009 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS -Apr-29 23:09:27.011 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-29 23:09:27.011 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-29 23:09:27.019 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:PREMERGE -Apr-29 23:09:27.022 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-29 23:09:27.022 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-29 23:09:27.040 [main] DEBUG nextflow.Session - Config process names validation disabled as requested -Apr-29 23:09:27.046 [main] DEBUG nextflow.Session - Igniting dataflow network (17) -Apr-29 23:09:27.052 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:FASTQC -Apr-29 23:09:27.055 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:MULTIQC -Apr-29 23:09:27.055 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX -Apr-29 23:09:27.057 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_MEM -Apr-29 23:09:27.057 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS -Apr-29 23:09:27.057 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:PREMERGE -Apr-29 23:09:27.057 [main] DEBUG nextflow.script.ScriptRunner - Parsed script files: +Apr-30 01:54:44.454 [main] DEBUG n.validation.ValidationExtension - Starting parameters validation +Apr-30 01:54:44.482 [main] WARN n.validation.ValidationExtension - The following invalid input values have been detected: + +* --reading_frame: 352-1383 + + +Apr-30 01:54:44.760 [main] DEBUG nextflow.validation.SchemaEvaluator - Started validating /BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv +Apr-30 01:54:45.826 [main] DEBUG nextflow.validation.SchemaEvaluator - Validation of file 'https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv' passed! +Apr-30 01:54:46.102 [main] DEBUG n.v.FormatDirectoryPathEvaluator - Cloud blob storage paths are not supported by 'FormatDirectoryPathEvaluator': 's3://ngi-igenomes/igenomes/' +Apr-30 01:54:46.105 [main] DEBUG n.validation.ValidationExtension - Finishing parameters validation +Apr-30 01:54:46.205 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:FASTQC +Apr-30 01:54:46.209 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:FASTQC` matches process NFCORE_DMSCORE:DMSCORE:FASTQC +Apr-30 01:54:46.225 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-30 01:54:46.225 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-30 01:54:46.233 [main] DEBUG nextflow.executor.Executor - [warm up] executor > local +Apr-30 01:54:46.240 [main] DEBUG n.processor.LocalPollingMonitor - Creating local task monitor for executor 'local' > cpus=8; memory=8 GB; capacity=8; pollInterval=100ms; dumpInterval=5m +Apr-30 01:54:46.243 [main] DEBUG n.processor.TaskPollingMonitor - >>> barrier register (monitor: local) +Apr-30 01:54:46.466 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:MULTIQC +Apr-30 01:54:46.467 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:MULTIQC` matches process NFCORE_DMSCORE:DMSCORE:MULTIQC +Apr-30 01:54:46.471 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-30 01:54:46.471 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-30 01:54:46.486 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BWA_INDEX +Apr-30 01:54:46.490 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-30 01:54:46.490 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-30 01:54:46.512 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_high` matches labels `process_high` for process with name NFCORE_DMSCORE:DMSCORE:BWA_MEM +Apr-30 01:54:46.518 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-30 01:54:46.518 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-30 01:54:46.534 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS +Apr-30 01:54:46.538 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-30 01:54:46.538 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-30 01:54:46.553 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:PREMERGE +Apr-30 01:54:46.558 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-30 01:54:46.558 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-30 01:54:46.574 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS +Apr-30 01:54:46.584 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +Apr-30 01:54:46.584 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +Apr-30 01:54:46.605 [main] DEBUG nextflow.Session - Config process names validation disabled as requested +Apr-30 01:54:46.608 [main] DEBUG nextflow.Session - Igniting dataflow network (20) +Apr-30 01:54:46.615 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:FASTQC +Apr-30 01:54:46.617 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:MULTIQC +Apr-30 01:54:46.617 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX +Apr-30 01:54:46.618 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_MEM +Apr-30 01:54:46.618 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS +Apr-30 01:54:46.618 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:PREMERGE +Apr-30 01:54:46.618 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS +Apr-30 01:54:46.619 [main] DEBUG nextflow.script.ScriptRunner - Parsed script files: Script_e5f965e09aa3641b: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/premerge.nf Script_113f045ba19c2c46: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/fastqc/main.nf + Script_daa9f53e0ae961a1: /Users/benjaminwehnert/dmscore/./workflows/dmscore.nf Script_4f612cfd8c52e8cd: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nextflow_pipeline/main.nf Script_215a636da7ab24a2: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfschema_plugin/main.nf Script_68606fccdecc54d9: /Users/benjaminwehnert/dmscore/./subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf Script_0f3077e98f6a93f6: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/mem/main.nf - Script_5c4e8d4051efa81e: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfcore_pipeline/main.nf + Script_5c4e8d4051efa81e: /Users/benjaminwehnert/dmscore/./subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfcore_pipeline/main.nf Script_4ff7366a79e0ef06: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/bamfilteringdms.nf - Script_5706c8cc12ed2c3e: /Users/benjaminwehnert/dmscore/./workflows/dmscore.nf Script_c568aae5b239c1c5: /Users/benjaminwehnert/dmscore/main.nf Script_4955288afd8ca61e: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/multiqc/main.nf + Script_a41bf2685e9281c5: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/gatk/saturationmutagenesis.nf Script_25aa31c18d513c61: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/index/main.nf -Apr-29 23:09:27.058 [main] DEBUG nextflow.script.ScriptRunner - > Awaiting termination -Apr-29 23:09:27.058 [main] DEBUG nextflow.Session - Session await -Apr-29 23:09:27.157 [Actor Thread 10] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.001 s; external sort time: 0.017 s; total time: 0.018 s -Apr-29 23:09:27.157 [Actor Thread 1] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.001 s; external sort time: 0.017 s; total time: 0.018 s -Apr-29 23:09:27.162 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/52ef908653c9bf719aa469cfe21f3a30 -Apr-29 23:09:27.162 [Actor Thread 10] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/7ce3707a79436715f6c7474bc3e3e7ce -Apr-29 23:09:27.173 [Actor Thread 10] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-10246409531653428819 -Apr-29 23:09:27.173 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-17840128110603128341 -Apr-29 23:09:27.180 [Actor Thread 4] DEBUG nextflow.util.HashBuilder - [WARN] Unknown hashing type: class Script_5706c8cc12ed2c3e$_runScript_closure1$_closure4 -Apr-29 23:09:27.261 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-29 23:09:27.263 [Task submitter] INFO nextflow.Session - [60/7b7f6b] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta) -Apr-29 23:09:27.524 [FileTransfer-1] DEBUG nextflow.file.FilePorter - Copying foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq to work dir: /Users/benjaminwehnert/dmscore/work/stage-f85367ca-5bb8-4421-882b-4615ea49f7a1/ca/48d11c2e7271ee34cc948495531277/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq -Apr-29 23:09:27.524 [FileTransfer-2] DEBUG nextflow.file.FilePorter - Copying foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq to work dir: /Users/benjaminwehnert/dmscore/work/stage-f85367ca-5bb8-4421-882b-4615ea49f7a1/74/ad28840a55d8285cd0b61430cb8ed1/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq -Apr-29 23:09:29.547 [Actor Thread 1] INFO nextflow.file.FilePorter - Staging foreign file: https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq -Apr-29 23:09:31.564 [Actor Thread 1] INFO nextflow.file.FilePorter - Staging foreign file: https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq -Apr-29 23:09:32.670 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 1; name: NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/60/7b7f6b5bf9b2a8a1c4462bed3dc193] -Apr-29 23:09:32.673 [Task monitor] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'TaskFinalizer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false -Apr-29 23:09:32.751 [TaskFinalizer-1] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'PublishDir' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false -Apr-29 23:09:32.898 [Actor Thread 7] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process -Apr-29 23:09:39.217 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-29 23:09:39.218 [Task submitter] INFO nextflow.Session - [17/9fd3ef] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_MEM (gid1a_1_quality_1_pe) -Apr-29 23:09:43.924 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 3; name: NFCORE_DMSCORE:DMSCORE:BWA_MEM (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/17/9fd3ef8a622a9fc8a8f5c8d1b8397c] -Apr-29 23:09:43.936 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-29 23:09:43.936 [Task submitter] INFO nextflow.Session - [ca/ae6468] Submitted process > NFCORE_DMSCORE:DMSCORE:FASTQC (gid1a_1_quality_1_pe) -Apr-29 23:09:44.084 [TaskFinalizer-2] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<1:1> -Apr-29 23:09:44.084 [TaskFinalizer-2] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<2:1> -Apr-29 23:09:44.084 [TaskFinalizer-2] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<3:1> -Apr-29 23:09:44.129 [Actor Thread 4] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process -Apr-29 23:09:54.512 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 2; name: NFCORE_DMSCORE:DMSCORE:FASTQC (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/ca/ae646873842dc826f95e58e0508b6d] -Apr-29 23:09:54.536 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-29 23:09:54.537 [Task submitter] INFO nextflow.Session - [c7/952c5e] Submitted process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS (gid1a_1_quality_1_pe) -Apr-29 23:09:54.642 [Actor Thread 4] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 2; slices: 1; internal sort time: 0.003 s; external sort time: 0.001 s; total time: 0.004 s -Apr-29 23:09:54.645 [Actor Thread 4] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/1172d3993855a8b4f3a6ae35d54320f1 -Apr-29 23:09:54.649 [Actor Thread 4] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-3409379234343285724 -Apr-29 23:09:58.685 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 4; name: NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/c7/952c5eaa65f9728c2a31229c8816fd] -Apr-29 23:09:58.694 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-29 23:09:58.695 [Task submitter] INFO nextflow.Session - [ab/48ed5b] Submitted process > NFCORE_DMSCORE:DMSCORE:MULTIQC -Apr-29 23:09:58.700 [Actor Thread 13] DEBUG nextflow.processor.TaskProcessor - Handling unexpected condition for - task: name=NFCORE_DMSCORE:DMSCORE:PREMERGE (1); work-dir=null - error [nextflow.exception.ProcessUnrecoverableException]: Not a valid path value type: Script_5706c8cc12ed2c3e$_runScript_closure1$_closure4 () -Apr-29 23:09:58.727 [Actor Thread 13] ERROR nextflow.processor.TaskProcessor - Error executing process > 'NFCORE_DMSCORE:DMSCORE:PREMERGE (1)' +Apr-30 01:54:46.620 [main] DEBUG nextflow.script.ScriptRunner - > Awaiting termination +Apr-30 01:54:46.620 [main] DEBUG nextflow.Session - Session await +Apr-30 01:54:46.774 [Actor Thread 12] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.002 s; external sort time: 0.022 s; total time: 0.024 s +Apr-30 01:54:46.774 [Actor Thread 5] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.002 s; external sort time: 0.022 s; total time: 0.024 s +Apr-30 01:54:46.776 [Actor Thread 12] DEBUG nextflow.file.FileCollector - >> temp file exists? false +Apr-30 01:54:46.776 [Actor Thread 5] DEBUG nextflow.file.FileCollector - >> temp file exists? false +Apr-30 01:54:46.777 [Actor Thread 5] DEBUG nextflow.file.FileCollector - Missed collect-file cache -- cause: java.nio.file.NoSuchFileException: /Users/benjaminwehnert/dmscore/work/collect-file/9e483aea9ab8efe250742af48b518fcb +Apr-30 01:54:46.777 [Actor Thread 12] DEBUG nextflow.file.FileCollector - Missed collect-file cache -- cause: java.nio.file.NoSuchFileException: /Users/benjaminwehnert/dmscore/work/collect-file/baea5eba38399e9f44aa2eb0cfb68beb +Apr-30 01:54:46.783 [Actor Thread 5] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/9e483aea9ab8efe250742af48b518fcb +Apr-30 01:54:46.784 [Actor Thread 12] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/baea5eba38399e9f44aa2eb0cfb68beb +Apr-30 01:54:46.798 [Actor Thread 5] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-3833846597375679323 +Apr-30 01:54:46.798 [Actor Thread 12] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-11497403584474485770 +Apr-30 01:54:46.799 [Actor Thread 14] DEBUG nextflow.util.HashBuilder - [WARN] Unknown hashing type: class Script_daa9f53e0ae961a1$_runScript_closure1$_closure6 +Apr-30 01:54:46.889 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-30 01:54:46.892 [Task submitter] INFO nextflow.Session - [63/e5edfa] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta) +Apr-30 01:54:47.399 [FileTransfer-2] DEBUG nextflow.file.FilePorter - Local cache found for foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq at /Users/benjaminwehnert/dmscore/work/stage-36458275-f468-409a-bb18-98996facae84/cc/f938a8775ac8befd4337b2732c7dc9/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq +Apr-30 01:54:47.399 [FileTransfer-1] DEBUG nextflow.file.FilePorter - Local cache found for foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq at /Users/benjaminwehnert/dmscore/work/stage-36458275-f468-409a-bb18-98996facae84/28/abb932c7e69700944b1605c1ee72d2/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq +Apr-30 01:54:47.445 [Actor Thread 4] INFO nextflow.processor.TaskProcessor - [9e/d2b28e] Cached process > NFCORE_DMSCORE:DMSCORE:FASTQC (gid1a_1_quality_1_pe) +Apr-30 01:54:47.483 [Actor Thread 4] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'PublishDir' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +Apr-30 01:54:47.515 [Actor Thread 5] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 2; slices: 1; internal sort time: 0.002 s; external sort time: 0.001 s; total time: 0.003 s +Apr-30 01:54:47.515 [Actor Thread 5] DEBUG nextflow.file.FileCollector - >> temp file exists? true +Apr-30 01:54:47.517 [Actor Thread 5] DEBUG nextflow.file.FileCollector - Retrieved cached collect-files from: /Users/benjaminwehnert/dmscore/work/collect-file/0dde6dd9bfe7393330209153ab0f204f +Apr-30 01:54:47.520 [Actor Thread 5] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-1489236878513704241 +Apr-30 01:54:51.271 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 1; name: NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/63/e5edfa2e469709a5513d0401c69f03] +Apr-30 01:54:51.275 [Task monitor] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'TaskFinalizer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +Apr-30 01:54:51.321 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-30 01:54:51.326 [Task submitter] INFO nextflow.Session - [2b/9ebde1] Submitted process > NFCORE_DMSCORE:DMSCORE:MULTIQC +Apr-30 01:54:51.410 [Actor Thread 12] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process +Apr-30 01:55:01.965 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 3; name: NFCORE_DMSCORE:DMSCORE:MULTIQC; status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/2b/9ebde1b8d8ace30edde0fba5d024f1] +Apr-30 01:55:02.002 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-30 01:55:02.003 [Task submitter] INFO nextflow.Session - [44/fb3934] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_MEM (gid1a_1_quality_1_pe) +Apr-30 01:55:02.148 [TaskFinalizer-2] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:MULTIQC > Skipping output binding because one or more optional files are missing: fileoutparam<2> +Apr-30 01:55:06.427 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 4; name: NFCORE_DMSCORE:DMSCORE:BWA_MEM (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/44/fb39348cda0e426f67e945b3b453c9] +Apr-30 01:55:06.450 [TaskFinalizer-3] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<1:1> +Apr-30 01:55:06.450 [TaskFinalizer-3] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<2:1> +Apr-30 01:55:06.450 [TaskFinalizer-3] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<3:1> +Apr-30 01:55:06.503 [Actor Thread 4] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process +Apr-30 01:55:06.516 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-30 01:55:06.517 [Task submitter] INFO nextflow.Session - [4f/32eeb6] Submitted process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS (gid1a_1_quality_1_pe) +Apr-30 01:55:10.625 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 5; name: NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/4f/32eeb651f29b281170859a5ccea65d] +Apr-30 01:55:10.698 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-30 01:55:10.698 [Task submitter] INFO nextflow.Session - [1f/f5a995] Submitted process > NFCORE_DMSCORE:DMSCORE:PREMERGE (gid1a_1_quality_1_pe) +Apr-30 01:55:13.152 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 6; name: NFCORE_DMSCORE:DMSCORE:PREMERGE (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/1f/f5a99582c50d984c21bf1d7f2ff192] +Apr-30 01:55:13.178 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +Apr-30 01:55:13.178 [Task submitter] INFO nextflow.Session - [97/8e2b73] Submitted process > NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe) +Apr-30 01:58:37.622 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 7; name: NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe); status: COMPLETED; exit: 127; error: -; workDir: /Users/benjaminwehnert/dmscore/work/97/8e2b7342dc0c54a5e5720f3f01e2e9] +Apr-30 01:58:37.686 [TaskFinalizer-6] DEBUG nextflow.processor.TaskProcessor - Handling unexpected condition for + task: name=NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe); work-dir=/Users/benjaminwehnert/dmscore/work/97/8e2b7342dc0c54a5e5720f3f01e2e9 + error [nextflow.exception.ProcessFailedException]: Process `NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe)` terminated with an error exit status (127) +Apr-30 01:58:37.742 [TaskFinalizer-6] ERROR nextflow.processor.TaskProcessor - Error executing process > 'NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe)' Caused by: - Not a valid path value type: Script_5706c8cc12ed2c3e$_runScript_closure1$_closure4 () - - - -Tip: you can try to figure out what's wrong by changing to the process work dir and showing the script file named `.command.sh` -Apr-29 23:09:58.729 [Actor Thread 13] INFO nextflow.Session - Execution cancelled -- Finishing pending tasks before exit -Apr-29 23:09:58.743 [Actor Thread 13] ERROR nextflow.Nextflow - Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting -Apr-29 23:09:58.744 [main] DEBUG nextflow.Session - Session await > all processes finished -Apr-29 23:09:58.748 [Actor Thread 10] DEBUG nextflow.processor.TaskProcessor - Handling unexpected condition for - task: name=NFCORE_DMSCORE:DMSCORE:PREMERGE; work-dir=null - error [java.lang.InterruptedException]: java.lang.InterruptedException -Apr-29 23:10:08.729 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 5; name: NFCORE_DMSCORE:DMSCORE:MULTIQC; status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/ab/48ed5bed903c8f8bdbc4507c19909f] -Apr-29 23:10:08.739 [Task monitor] DEBUG n.processor.TaskPollingMonitor - <<< barrier arrives (monitor: local) - terminating tasks monitor poll loop -Apr-29 23:10:08.740 [main] DEBUG nextflow.Session - Session await > all barriers passed -Apr-29 23:10:08.766 [TaskFinalizer-5] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:MULTIQC > Skipping output binding because one or more optional files are missing: fileoutparam<2> -Apr-29 23:10:08.802 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'TaskFinalizer' shutdown completed (hard=false) -Apr-29 23:10:08.804 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'PublishDir' shutdown completed (hard=false) -Apr-29 23:10:08.818 [main] INFO nextflow.Nextflow - -[nf-core/dmscore] Pipeline completed with errors- -Apr-29 23:10:08.826 [main] DEBUG n.trace.WorkflowStatsObserver - Workflow completed > WorkflowStats[succeededCount=5; failedCount=0; ignoredCount=0; cachedCount=0; pendingCount=0; submittedCount=0; runningCount=0; retriesCount=0; abortedCount=0; succeedDuration=1m 1s; failedDuration=0ms; cachedDuration=0ms;loadCpus=0; loadMemory=0; peakRunning=2; peakCpus=8; peakMemory=16 GB; ] -Apr-29 23:10:08.826 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow completed -- saving trace file -Apr-29 23:10:08.835 [main] DEBUG nextflow.trace.ReportObserver - Workflow completed -- rendering execution report -Apr-29 23:10:10.258 [main] DEBUG nextflow.trace.TimelineObserver - Workflow completed -- rendering execution timeline -Apr-29 23:10:10.573 [main] DEBUG nextflow.cache.CacheDB - Closing CacheDB done -Apr-29 23:10:10.593 [main] INFO org.pf4j.AbstractPluginManager - Stop plugin 'nf-schema@2.3.0' -Apr-29 23:10:10.593 [main] DEBUG nextflow.plugin.BasePlugin - Plugin stopped nf-schema -Apr-29 23:10:10.594 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'FileTransfer' shutdown completed (hard=false) -Apr-29 23:10:10.597 [main] DEBUG nextflow.script.ScriptRunner - > Execution complete -- Goodbye + Process `NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe)` terminated with an error exit status (127) + + +Command executed: + + # Index reference + samtools faidx GID1A_SUNi_ref_small.fasta + gatk CreateSequenceDictionary -R GID1A_SUNi_ref_small.fasta + + # Read start and stop codon from input + start_stop_codon="352-1383" + + # Run GATK AnalyzeSaturationMutagenesis + gatk AnalyzeSaturationMutagenesis -I merged_reads.bam -R GID1A_SUNi_ref_small.fasta --orf $start_stop_codon --paired-mode false --min-q 30 --min-variant-obs 3 -O gatk_output + + # Save versions + cat <<-END_VERSIONS > versions.yml + "NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS": + samtools: $(samtools --version |& sed '1!d ; s/samtools //') + gatk: $(gatk --version |& sed 's/^.*GATK//') + END_VERSIONS + +Command exit status: + 127 + +Command output: + (empty) + +Command error: + Unable to find image 'community.wave.seqera.io/library/gatk_samtools_java-1.7.0-openjdk-conda-aarch64:50bbe1742b62bc8f' locally + 50bbe1742b62bc8f: Pulling from library/gatk_samtools_java-1.7.0-openjdk-conda-aarch64 + b9cdf96edaf9: Pulling fs layer + 3e5b316ff8d6: Pulling fs layer + b9cdf96edaf9: Download complete + 3e5b316ff8d6: Download complete + Digest: sha256:ab1b440a72d9caec2beb4d205294eb7d43c33448ccb863597ae6858674718d69 + Status: Downloaded newer image for community.wave.seqera.io/library/gatk_samtools_java-1.7.0-openjdk-conda-aarch64:50bbe1742b62bc8f + .command.sh: line 10: gatk: command not found + +Work dir: + /Users/benjaminwehnert/dmscore/work/97/8e2b7342dc0c54a5e5720f3f01e2e9 + +Tip: when you have fixed the problem you can continue the execution adding the option `-resume` to the run command line +Apr-30 01:58:37.755 [main] DEBUG nextflow.Session - Session await > all processes finished +Apr-30 01:58:37.779 [TaskFinalizer-6] INFO nextflow.Session - Execution cancelled -- Finishing pending tasks before exit +Apr-30 01:58:37.797 [Task monitor] DEBUG n.processor.TaskPollingMonitor - <<< barrier arrives (monitor: local) - terminating tasks monitor poll loop +Apr-30 01:58:37.822 [main] DEBUG nextflow.Session - Session await > all barriers passed +Apr-30 01:58:37.883 [TaskFinalizer-6] ERROR nextflow.Nextflow - Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting +Apr-30 01:58:37.884 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'TaskFinalizer' shutdown completed (hard=false) +Apr-30 01:58:37.886 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'PublishDir' shutdown completed (hard=false) +Apr-30 01:58:37.900 [main] INFO nextflow.Nextflow - -[nf-core/dmscore] Pipeline completed with errors- +Apr-30 01:58:37.908 [main] DEBUG n.trace.WorkflowStatsObserver - Workflow completed > WorkflowStats[succeededCount=5; failedCount=1; ignoredCount=0; cachedCount=1; pendingCount=0; submittedCount=0; runningCount=0; retriesCount=0; abortedCount=0; succeedDuration=24.5s; failedDuration=13m 37s; cachedDuration=48s;loadCpus=0; loadMemory=0; peakRunning=2; peakCpus=5; peakMemory=14 GB; ] +Apr-30 01:58:37.909 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow completed -- saving trace file +Apr-30 01:58:37.912 [main] DEBUG nextflow.trace.ReportObserver - Workflow completed -- rendering execution report +Apr-30 01:58:39.474 [main] DEBUG nextflow.trace.TimelineObserver - Workflow completed -- rendering execution timeline +Apr-30 01:58:39.678 [main] DEBUG nextflow.cache.CacheDB - Closing CacheDB done +Apr-30 01:58:39.701 [main] INFO org.pf4j.AbstractPluginManager - Stop plugin 'nf-schema@2.3.0' +Apr-30 01:58:39.701 [main] DEBUG nextflow.plugin.BasePlugin - Plugin stopped nf-schema +Apr-30 01:58:39.702 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'FileTransfer' shutdown completed (hard=false) +Apr-30 01:58:39.704 [main] DEBUG nextflow.script.ScriptRunner - > Execution complete -- Goodbye diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf new file mode 100644 index 0000000..09a49ed --- /dev/null +++ b/modules/local/dmsanalysis/aaseq.nf @@ -0,0 +1,91 @@ +// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) +// https://github.com/nf-core/modules/tree/master/modules/nf-core/ +// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: +// https://nf-co.re/join +// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. +// All other parameters MUST be provided using the "task.ext" directive, see here: +// https://www.nextflow.io/docs/latest/process.html#ext +// where "task.ext" is a string. +// Any parameters that need to be evaluated in the context of a particular sample +// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. +// TODO nf-core: Software that can be piped together SHOULD be added to separate module files +// unless there is a run-time, storage advantage in implementing in this way +// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: +// bwa mem | samtools view -B -T ref.fasta +// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty +// list (`[]`) instead of a file can be used to work around this issue. + +process DMSANALYSIS_AASEQ { + tag "$meta.id" + label 'process_low' + + // TODO nf-core: List required Conda package(s). + // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). + // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. + // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/YOUR-TOOL-HERE': + 'biocontainers/YOUR-TOOL-HERE' }" + + input: + // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" + // MUST be provided as an input via a Groovy Map called "meta". + // This information may not be required in some instances e.g. indexing reference genome files: + // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf + // TODO nf-core: Where applicable please provide/convert compressed files as input/output + // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. + tuple val(meta), path(bam) + + output: + // TODO nf-core: Named file extensions MUST be emitted for ALL output channels + tuple val(meta), path("*.bam"), emit: bam + // TODO nf-core: List additional required output channels/values here + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 + // If the software is unable to output a version number on the command-line then it can be manually specified + // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf + // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) + // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive + // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter + // using the Nextflow "task" variable e.g. "--threads $task.cpus" + // TODO nf-core: Please replace the example samtools command below with your module's command + // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) + """ + samtools \\ + sort \\ + $args \\ + -@ $task.cpus \\ + -o ${prefix}.bam \\ + -T $prefix \\ + $bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ + + stub: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: A stub section should mimic the execution of the original module as best as possible + // Have a look at the following examples: + // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 + // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 + """ + touch ${prefix}.bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ +} diff --git a/modules/local/dmsanalysis/annotategatk.nf b/modules/local/dmsanalysis/annotategatk.nf new file mode 100644 index 0000000..eac3c50 --- /dev/null +++ b/modules/local/dmsanalysis/annotategatk.nf @@ -0,0 +1,91 @@ +// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) +// https://github.com/nf-core/modules/tree/master/modules/nf-core/ +// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: +// https://nf-co.re/join +// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. +// All other parameters MUST be provided using the "task.ext" directive, see here: +// https://www.nextflow.io/docs/latest/process.html#ext +// where "task.ext" is a string. +// Any parameters that need to be evaluated in the context of a particular sample +// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. +// TODO nf-core: Software that can be piped together SHOULD be added to separate module files +// unless there is a run-time, storage advantage in implementing in this way +// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: +// bwa mem | samtools view -B -T ref.fasta +// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty +// list (`[]`) instead of a file can be used to work around this issue. + +process DMSANALYSIS_ANNOTATEGATK { + tag "$meta.id" + label 'process_single' + + // TODO nf-core: List required Conda package(s). + // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). + // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. + // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/YOUR-TOOL-HERE': + 'biocontainers/YOUR-TOOL-HERE' }" + + input: + // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" + // MUST be provided as an input via a Groovy Map called "meta". + // This information may not be required in some instances e.g. indexing reference genome files: + // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf + // TODO nf-core: Where applicable please provide/convert compressed files as input/output + // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. + tuple val(meta), path(bam) + + output: + // TODO nf-core: Named file extensions MUST be emitted for ALL output channels + tuple val(meta), path("*.bam"), emit: bam + // TODO nf-core: List additional required output channels/values here + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 + // If the software is unable to output a version number on the command-line then it can be manually specified + // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf + // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) + // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive + // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter + // using the Nextflow "task" variable e.g. "--threads $task.cpus" + // TODO nf-core: Please replace the example samtools command below with your module's command + // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) + """ + samtools \\ + sort \\ + $args \\ + -@ $task.cpus \\ + -o ${prefix}.bam \\ + -T $prefix \\ + $bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ + + stub: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: A stub section should mimic the execution of the original module as best as possible + // Have a look at the following examples: + // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 + // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 + """ + touch ${prefix}.bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ +} diff --git a/modules/local/dmsanalysis/visualization.nf b/modules/local/dmsanalysis/visualization.nf new file mode 100644 index 0000000..7ddcc83 --- /dev/null +++ b/modules/local/dmsanalysis/visualization.nf @@ -0,0 +1,91 @@ +// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) +// https://github.com/nf-core/modules/tree/master/modules/nf-core/ +// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: +// https://nf-co.re/join +// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. +// All other parameters MUST be provided using the "task.ext" directive, see here: +// https://www.nextflow.io/docs/latest/process.html#ext +// where "task.ext" is a string. +// Any parameters that need to be evaluated in the context of a particular sample +// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. +// TODO nf-core: Software that can be piped together SHOULD be added to separate module files +// unless there is a run-time, storage advantage in implementing in this way +// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: +// bwa mem | samtools view -B -T ref.fasta +// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty +// list (`[]`) instead of a file can be used to work around this issue. + +process DMSANALYSIS_VISUALIZATION { + tag "$meta.id" + label 'process_single' + + // TODO nf-core: List required Conda package(s). + // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). + // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. + // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/YOUR-TOOL-HERE': + 'biocontainers/YOUR-TOOL-HERE' }" + + input: + // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" + // MUST be provided as an input via a Groovy Map called "meta". + // This information may not be required in some instances e.g. indexing reference genome files: + // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf + // TODO nf-core: Where applicable please provide/convert compressed files as input/output + // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. + tuple val(meta), path(bam) + + output: + // TODO nf-core: Named file extensions MUST be emitted for ALL output channels + tuple val(meta), path("*.bam"), emit: bam + // TODO nf-core: List additional required output channels/values here + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 + // If the software is unable to output a version number on the command-line then it can be manually specified + // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf + // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) + // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive + // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter + // using the Nextflow "task" variable e.g. "--threads $task.cpus" + // TODO nf-core: Please replace the example samtools command below with your module's command + // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) + """ + samtools \\ + sort \\ + $args \\ + -@ $task.cpus \\ + -o ${prefix}.bam \\ + -T $prefix \\ + $bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ + + stub: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: A stub section should mimic the execution of the original module as best as possible + // Have a look at the following examples: + // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 + // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 + """ + touch ${prefix}.bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ +} From a1223df05aaf0528744495c9c2a1d57757821919 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 15:09:04 +0200 Subject: [PATCH 078/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 81 ++++++------------------------ 1 file changed, 16 insertions(+), 65 deletions(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index 09a49ed..43e639c 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -1,91 +1,42 @@ -// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) -// https://github.com/nf-core/modules/tree/master/modules/nf-core/ -// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: -// https://nf-co.re/join -// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. -// All other parameters MUST be provided using the "task.ext" directive, see here: -// https://www.nextflow.io/docs/latest/process.html#ext -// where "task.ext" is a string. -// Any parameters that need to be evaluated in the context of a particular sample -// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. -// TODO nf-core: Software that can be piped together SHOULD be added to separate module files -// unless there is a run-time, storage advantage in implementing in this way -// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: -// bwa mem | samtools view -B -T ref.fasta -// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty -// list (`[]`) instead of a file can be used to work around this issue. - process DMSANALYSIS_AASEQ { tag "$meta.id" - label 'process_low' + label 'process_single' - // TODO nf-core: List required Conda package(s). - // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). - // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. - // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. - conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/YOUR-TOOL-HERE': - 'biocontainers/YOUR-TOOL-HERE' }" + // Define the correct container and/or conda environment + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:8fa94107068d5af9" input: - // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" - // MUST be provided as an input via a Groovy Map called "meta". - // This information may not be required in some instances e.g. indexing reference genome files: - // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf - // TODO nf-core: Where applicable please provide/convert compressed files as input/output - // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. - tuple val(meta), path(bam) + tuple val(meta), path(wt_seq) + path pos_range output: - // TODO nf-core: Named file extensions MUST be emitted for ALL output channels - tuple val(meta), path("*.bam"), emit: bam - // TODO nf-core: List additional required output channels/values here - path "versions.yml" , emit: versions + tuple val(meta), path("aa_seq.txt"), emit: aa_seq + path "versions.yml", emit: versions when: task.ext.when == null || task.ext.when script: - def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 - // If the software is unable to output a version number on the command-line then it can be manually specified - // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf - // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) - // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive - // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter - // using the Nextflow "task" variable e.g. "--threads $task.cpus" - // TODO nf-core: Please replace the example samtools command below with your module's command - // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) """ - samtools \\ - sort \\ - $args \\ - -@ $task.cpus \\ - -o ${prefix}.bam \\ - -T $prefix \\ - $bam + start_stop_codon=\$(cat $pos_range) + + Rscript -e "source('../bin/aa_seq.R'); aa_seq('$wt_seq', '\$start_stop_codon', 'aa_seq.txt')" cat <<-END_VERSIONS > versions.yml - "${task.process}": - dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + "\${task.process}": + r: \$(R --version | head -n 1 | sed 's/^R version //') + aa_seq_script: custom_R_script_bin END_VERSIONS """ stub: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: A stub section should mimic the execution of the original module as best as possible - // Have a look at the following examples: - // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 - // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 """ - touch ${prefix}.bam - + touch aa_seq.txt + cat <<-END_VERSIONS > versions.yml "${task.process}": - dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + dmsanalysis_aaseq: dummy_version END_VERSIONS """ } From c82910aacd9b993ff9c7be6226b4e9ec41a8dec9 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Fri, 2 May 2025 15:10:56 +0200 Subject: [PATCH 079/317] R-scripts --- log.txt | 270 ------------------ .../dmsanalysis/bin/SeqDepth_simulation.R | 131 +++++++++ modules/local/dmsanalysis/bin/aa_seq.R | 37 +++ .../bin/complete_prefiltered_gatk.R | 79 +++++ .../local/dmsanalysis/bin/counts_heatmap.R | 154 ++++++++++ .../dmsanalysis/bin/counts_per_cov_heatmap.R | 154 ++++++++++ modules/local/dmsanalysis/bin/detect_codons.R | 65 +++++ .../bin/filter_gatk_by_codon_library.R | 124 ++++++++ .../local/dmsanalysis/bin/fitness_heatmap.R | 160 +++++++++++ .../local/dmsanalysis/bin/gatk_to_dimsum.R | 180 ++++++++++++ ...osition_biases_counts_and_counts_per_cov.R | 163 +++++++++++ .../bin/global_position_biases_cov.R | 86 ++++++ .../local/dmsanalysis/bin/install_packages.R | 82 ++++++ modules/local/dmsanalysis/bin/logdiff.R | 149 ++++++++++ .../dmsanalysis/bin/low_count_variants.R | 130 +++++++++ .../dmsanalysis/bin/possible_mutations.R | 105 +++++++ .../prepare_gatk_data_for_count_heatmaps.R | 82 ++++++ .../prepare_gatk_data_for_fitness_heatmap.R | 134 +++++++++ .../local/dmsanalysis/bin/process_raw_gatk.R | 46 +++ 19 files changed, 2061 insertions(+), 270 deletions(-) delete mode 100644 log.txt create mode 100644 modules/local/dmsanalysis/bin/SeqDepth_simulation.R create mode 100644 modules/local/dmsanalysis/bin/aa_seq.R create mode 100644 modules/local/dmsanalysis/bin/complete_prefiltered_gatk.R create mode 100644 modules/local/dmsanalysis/bin/counts_heatmap.R create mode 100644 modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R create mode 100644 modules/local/dmsanalysis/bin/detect_codons.R create mode 100644 modules/local/dmsanalysis/bin/filter_gatk_by_codon_library.R create mode 100644 modules/local/dmsanalysis/bin/fitness_heatmap.R create mode 100644 modules/local/dmsanalysis/bin/gatk_to_dimsum.R create mode 100644 modules/local/dmsanalysis/bin/global_position_biases_counts_and_counts_per_cov.R create mode 100644 modules/local/dmsanalysis/bin/global_position_biases_cov.R create mode 100644 modules/local/dmsanalysis/bin/install_packages.R create mode 100644 modules/local/dmsanalysis/bin/logdiff.R create mode 100644 modules/local/dmsanalysis/bin/low_count_variants.R create mode 100644 modules/local/dmsanalysis/bin/possible_mutations.R create mode 100644 modules/local/dmsanalysis/bin/prepare_gatk_data_for_count_heatmaps.R create mode 100644 modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R create mode 100644 modules/local/dmsanalysis/bin/process_raw_gatk.R diff --git a/log.txt b/log.txt deleted file mode 100644 index 59304d4..0000000 --- a/log.txt +++ /dev/null @@ -1,270 +0,0 @@ -Apr-30 01:54:39.166 [main] DEBUG nextflow.cli.Launcher - $> nextflow run . -profile test,docker --fasta /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta --reading_frame 352-1383 --outdir ./results -resume -Apr-30 01:54:39.306 [main] DEBUG nextflow.cli.CmdRun - N E X T F L O W ~ version 24.04.4 -Apr-30 01:54:39.333 [main] DEBUG nextflow.plugin.PluginsFacade - Setting up plugin manager > mode=prod; embedded=false; plugins-dir=/Users/benjaminwehnert/.nextflow/plugins; core-plugins: nf-amazon@2.5.3,nf-azure@1.6.1,nf-cloudcache@0.4.1,nf-codecommit@0.2.1,nf-console@1.1.3,nf-ga4gh@1.3.0,nf-google@1.13.2-patch1,nf-tower@1.9.1,nf-wave@1.4.2-patch1 -Apr-30 01:54:39.345 [main] INFO o.pf4j.DefaultPluginStatusProvider - Enabled plugins: [] -Apr-30 01:54:39.346 [main] INFO o.pf4j.DefaultPluginStatusProvider - Disabled plugins: [] -Apr-30 01:54:39.349 [main] INFO org.pf4j.DefaultPluginManager - PF4J version 3.12.0 in 'deployment' mode -Apr-30 01:54:39.366 [main] INFO org.pf4j.AbstractPluginManager - No plugins -Apr-30 01:54:40.248 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` -Apr-30 01:54:40.266 [main] DEBUG nextflow.config.ConfigBuilder - Found config local: /Users/benjaminwehnert/dmscore/nextflow.config -Apr-30 01:54:40.274 [main] DEBUG nextflow.config.ConfigBuilder - Parsing config file: /Users/benjaminwehnert/dmscore/nextflow.config -Apr-30 01:54:40.290 [main] DEBUG n.secret.LocalSecretsProvider - Secrets store: /Users/benjaminwehnert/.nextflow/secrets/store.json -Apr-30 01:54:40.293 [main] DEBUG nextflow.secret.SecretsLoader - Discovered secrets providers: [nextflow.secret.LocalSecretsProvider@606f81b5] - activable => nextflow.secret.LocalSecretsProvider@606f81b5 -Apr-30 01:54:40.304 [main] DEBUG nextflow.config.ConfigBuilder - Applying config profile: `test,docker` -Apr-30 01:54:41.801 [main] DEBUG nextflow.config.ConfigBuilder - Available config profiles: [bih, cfc_dev, uzl_omics, ifb_core, embl_hd, denbi_qbic, alice, mjolnir_globe, uppmax, giga, incliva, ilifu, ki_luria, uge, icr_alma, rosalind_uge, lugh, mccleary, unibe_ibu, vai, czbiohub_aws, jax, roslin, ccga_med, tes, scw, unc_longleaf, tigem, tubingen_apg, google, apollo, ipop_up, vsc_calcua, pdc_kth, googlels, ceci_nic5, humantechnopole, stjude, daisybio, eddie, medair, biowulf, apptainer, bi, bigpurple, adcra, cedars, pawsey_setonix, vsc_kul_uhasselt, pawsey_nimbus, ucl_myriad, utd_ganymede, charliecloud, seattlechildrens, icr_davros, ceres, arm, munin, rosalind, hasta, cfc, uzh, shu_bmrc, ebi_codon_slurm, ebc, ccga_dx, crick, ku_sund_danhead, marvin, shifter, biohpc_gen, mana, mamba, york_viking, unc_lccc, wehi, awsbatch, wustl_htcf, arcc, ceci_dragon2, imperial, maestro, software_license, cannon, genotoul, nci_gadi, abims, janelia, nu_genomics, googlebatch, oist, sahmri, kaust, alliance_canada, mpcdf, leicester, vsc_ugent, create, sage, cambridge, jex, podman, ebi_codon, cheaha, xanadu, nyu_hpc, test, marjorie, computerome, ucd_sonic, seg_globe, sanger, dkfz, bluebear, pasteur, einstein, ethz_euler, m3c, test_full, imb, ucl_cscluster, tuos_stanage, azurebatch, hki, seadragon, crukmi, csiro_petrichor, qmul_apocrita, wave, docker, engaging, gis, hypatia, psmn, eva, unity, cropdiversityhpc, nygc, fgcz, conda, crg, singularity, mpcdf_viper, pe2, self_hosted_runner, tufts, uw_hyak_pedslabs, binac2, debug, genouest, cbe, unsw_katana, gitpod, phoenix, seawulf, uod_hpc, fub_curta, uct_hpc, aws_tower, binac, fsu_draco] -Apr-30 01:54:41.868 [main] DEBUG nextflow.cli.CmdRun - Applied DSL=2 by global default -Apr-30 01:54:41.893 [main] DEBUG nextflow.cli.CmdRun - Launching `./main.nf` [jolly_cori] DSL2 - revision: 84101fc51c -Apr-30 01:54:41.895 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins declared=[nf-schema@2.3.0] -Apr-30 01:54:41.895 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins default=[] -Apr-30 01:54:41.895 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins resolved requirement=[nf-schema@2.3.0] -Apr-30 01:54:41.896 [main] DEBUG nextflow.plugin.PluginUpdater - Installing plugin nf-schema version: 2.3.0 -Apr-30 01:54:41.905 [main] INFO org.pf4j.AbstractPluginManager - Plugin 'nf-schema@2.3.0' resolved -Apr-30 01:54:41.906 [main] INFO org.pf4j.AbstractPluginManager - Start plugin 'nf-schema@2.3.0' -Apr-30 01:54:41.913 [main] DEBUG nextflow.plugin.BasePlugin - Plugin started nf-schema@2.3.0 -Apr-30 01:54:41.988 [main] DEBUG nextflow.Session - Session UUID: 36458275-f468-409a-bb18-98996facae84 -Apr-30 01:54:41.989 [main] DEBUG nextflow.Session - Run name: jolly_cori -Apr-30 01:54:41.989 [main] DEBUG nextflow.Session - Executor pool size: 8 -Apr-30 01:54:41.999 [main] DEBUG nextflow.file.FilePorter - File porter settings maxRetries=3; maxTransfers=50; pollTimeout=null -Apr-30 01:54:42.005 [main] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'FileTransfer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false -Apr-30 01:54:42.029 [main] DEBUG nextflow.cli.CmdRun - - Version: 24.04.4 build 5917 - Created: 01-08-2024 07:05 UTC (09:05 CEST) - System: Mac OS X 15.0 - Runtime: Groovy 4.0.21 on OpenJDK 64-Bit Server VM 17.0.13+0 - Encoding: UTF-8 (UTF-8) - Process: 3502@MacBook-Air-von-Benjamin.local [127.0.0.1] - CPUs: 8 - Mem: 8 GB (138.6 MB) - Swap: 8 GB (790.1 MB) -Apr-30 01:54:42.059 [main] DEBUG nextflow.Session - Work-dir: /Users/benjaminwehnert/dmscore/work [Mac OS X] -Apr-30 01:54:42.060 [main] DEBUG nextflow.Session - Script base path does not exist or is not a directory: /Users/benjaminwehnert/dmscore/bin -Apr-30 01:54:42.073 [main] DEBUG nextflow.executor.ExecutorFactory - Extension executors providers=[] -Apr-30 01:54:42.083 [main] DEBUG nextflow.Session - Observer factory: DefaultObserverFactory -Apr-30 01:54:42.105 [main] DEBUG nextflow.Session - Observer factory: ValidationObserverFactory -Apr-30 01:54:42.139 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` -Apr-30 01:54:42.164 [main] DEBUG nextflow.cache.CacheFactory - Using Nextflow cache factory: nextflow.cache.DefaultCacheFactory -Apr-30 01:54:42.172 [main] DEBUG nextflow.util.CustomThreadPool - Creating default thread pool > poolSize: 9; maxThreads: 1000 -Apr-30 01:54:42.275 [main] DEBUG nextflow.Session - Session start -Apr-30 01:54:42.276 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow started -- trace file: /Users/benjaminwehnert/dmscore/results/pipeline_info/execution_trace_2025-04-30_01-54-40.txt -Apr-30 01:54:42.437 [main] DEBUG nextflow.script.ScriptRunner - > Launching execution -Apr-30 01:54:43.415 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema -Apr-30 01:54:43.844 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema -Apr-30 01:54:43.845 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema -Apr-30 01:54:43.847 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema -Apr-30 01:54:43.848 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema -Apr-30 01:54:44.121 [main] WARN nextflow.script.ScriptBinding - Access to undefined parameter `min_counts` -- Initialise it to a default value eg. `params.min_counts = some_value` -Apr-30 01:54:44.245 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema -Apr-30 01:54:44.246 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema -Apr-30 01:54:44.248 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema -Apr-30 01:54:44.249 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema -Apr-30 01:54:44.452 [main] INFO nextflow.Nextflow - ------------------------------------------------------- - ,--./,-. - ___ __ __ __ ___ /,-._.--~' - |\ | |__ __ / ` / \ |__) |__ } { - | \| | \__, \__/ | \ |___ \`-._,-`-, - `._,._,' - nf-core/dmscore 1.0.0dev ------------------------------------------------------- -Input/output options - input : https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv - outdir : ./results - -Reference genome options - genome : R64-1-1 - fasta : /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta - -Institutional config options - config_profile_name : Test profile - config_profile_description: Minimal test dataset to check pipeline function - -Generic options - trace_report_suffix : 2025-04-30_01-54-40 - -Core Nextflow options - runName : jolly_cori - containerEngine : docker - launchDir : /Users/benjaminwehnert/dmscore - workDir : /Users/benjaminwehnert/dmscore/work - projectDir : /Users/benjaminwehnert/dmscore - userName : benjaminwehnert - profile : test,docker - configFiles : /Users/benjaminwehnert/dmscore/nextflow.config - -!! Only displaying parameters that differ from the pipeline defaults !! ------------------------------------------------------- -* The nf-core framework - https://doi.org/10.1038/s41587-020-0439-x - -* Software dependencies - https://github.com/nf-core/dmscore/blob/master/CITATIONS.md - -Apr-30 01:54:44.454 [main] DEBUG n.validation.ValidationExtension - Starting parameters validation -Apr-30 01:54:44.482 [main] WARN n.validation.ValidationExtension - The following invalid input values have been detected: - -* --reading_frame: 352-1383 - - -Apr-30 01:54:44.760 [main] DEBUG nextflow.validation.SchemaEvaluator - Started validating /BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv -Apr-30 01:54:45.826 [main] DEBUG nextflow.validation.SchemaEvaluator - Validation of file 'https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv' passed! -Apr-30 01:54:46.102 [main] DEBUG n.v.FormatDirectoryPathEvaluator - Cloud blob storage paths are not supported by 'FormatDirectoryPathEvaluator': 's3://ngi-igenomes/igenomes/' -Apr-30 01:54:46.105 [main] DEBUG n.validation.ValidationExtension - Finishing parameters validation -Apr-30 01:54:46.205 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:FASTQC -Apr-30 01:54:46.209 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:FASTQC` matches process NFCORE_DMSCORE:DMSCORE:FASTQC -Apr-30 01:54:46.225 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-30 01:54:46.225 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-30 01:54:46.233 [main] DEBUG nextflow.executor.Executor - [warm up] executor > local -Apr-30 01:54:46.240 [main] DEBUG n.processor.LocalPollingMonitor - Creating local task monitor for executor 'local' > cpus=8; memory=8 GB; capacity=8; pollInterval=100ms; dumpInterval=5m -Apr-30 01:54:46.243 [main] DEBUG n.processor.TaskPollingMonitor - >>> barrier register (monitor: local) -Apr-30 01:54:46.466 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:MULTIQC -Apr-30 01:54:46.467 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:MULTIQC` matches process NFCORE_DMSCORE:DMSCORE:MULTIQC -Apr-30 01:54:46.471 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-30 01:54:46.471 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-30 01:54:46.486 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BWA_INDEX -Apr-30 01:54:46.490 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-30 01:54:46.490 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-30 01:54:46.512 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_high` matches labels `process_high` for process with name NFCORE_DMSCORE:DMSCORE:BWA_MEM -Apr-30 01:54:46.518 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-30 01:54:46.518 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-30 01:54:46.534 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS -Apr-30 01:54:46.538 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-30 01:54:46.538 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-30 01:54:46.553 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:PREMERGE -Apr-30 01:54:46.558 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-30 01:54:46.558 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-30 01:54:46.574 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS -Apr-30 01:54:46.584 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -Apr-30 01:54:46.584 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -Apr-30 01:54:46.605 [main] DEBUG nextflow.Session - Config process names validation disabled as requested -Apr-30 01:54:46.608 [main] DEBUG nextflow.Session - Igniting dataflow network (20) -Apr-30 01:54:46.615 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:FASTQC -Apr-30 01:54:46.617 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:MULTIQC -Apr-30 01:54:46.617 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX -Apr-30 01:54:46.618 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_MEM -Apr-30 01:54:46.618 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS -Apr-30 01:54:46.618 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:PREMERGE -Apr-30 01:54:46.618 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS -Apr-30 01:54:46.619 [main] DEBUG nextflow.script.ScriptRunner - Parsed script files: - Script_e5f965e09aa3641b: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/premerge.nf - Script_113f045ba19c2c46: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/fastqc/main.nf - Script_daa9f53e0ae961a1: /Users/benjaminwehnert/dmscore/./workflows/dmscore.nf - Script_4f612cfd8c52e8cd: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nextflow_pipeline/main.nf - Script_215a636da7ab24a2: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfschema_plugin/main.nf - Script_68606fccdecc54d9: /Users/benjaminwehnert/dmscore/./subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf - Script_0f3077e98f6a93f6: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/mem/main.nf - Script_5c4e8d4051efa81e: /Users/benjaminwehnert/dmscore/./subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfcore_pipeline/main.nf - Script_4ff7366a79e0ef06: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/bamfilteringdms.nf - Script_c568aae5b239c1c5: /Users/benjaminwehnert/dmscore/main.nf - Script_4955288afd8ca61e: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/multiqc/main.nf - Script_a41bf2685e9281c5: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/gatk/saturationmutagenesis.nf - Script_25aa31c18d513c61: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/index/main.nf -Apr-30 01:54:46.620 [main] DEBUG nextflow.script.ScriptRunner - > Awaiting termination -Apr-30 01:54:46.620 [main] DEBUG nextflow.Session - Session await -Apr-30 01:54:46.774 [Actor Thread 12] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.002 s; external sort time: 0.022 s; total time: 0.024 s -Apr-30 01:54:46.774 [Actor Thread 5] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.002 s; external sort time: 0.022 s; total time: 0.024 s -Apr-30 01:54:46.776 [Actor Thread 12] DEBUG nextflow.file.FileCollector - >> temp file exists? false -Apr-30 01:54:46.776 [Actor Thread 5] DEBUG nextflow.file.FileCollector - >> temp file exists? false -Apr-30 01:54:46.777 [Actor Thread 5] DEBUG nextflow.file.FileCollector - Missed collect-file cache -- cause: java.nio.file.NoSuchFileException: /Users/benjaminwehnert/dmscore/work/collect-file/9e483aea9ab8efe250742af48b518fcb -Apr-30 01:54:46.777 [Actor Thread 12] DEBUG nextflow.file.FileCollector - Missed collect-file cache -- cause: java.nio.file.NoSuchFileException: /Users/benjaminwehnert/dmscore/work/collect-file/baea5eba38399e9f44aa2eb0cfb68beb -Apr-30 01:54:46.783 [Actor Thread 5] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/9e483aea9ab8efe250742af48b518fcb -Apr-30 01:54:46.784 [Actor Thread 12] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/baea5eba38399e9f44aa2eb0cfb68beb -Apr-30 01:54:46.798 [Actor Thread 5] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-3833846597375679323 -Apr-30 01:54:46.798 [Actor Thread 12] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-11497403584474485770 -Apr-30 01:54:46.799 [Actor Thread 14] DEBUG nextflow.util.HashBuilder - [WARN] Unknown hashing type: class Script_daa9f53e0ae961a1$_runScript_closure1$_closure6 -Apr-30 01:54:46.889 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-30 01:54:46.892 [Task submitter] INFO nextflow.Session - [63/e5edfa] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta) -Apr-30 01:54:47.399 [FileTransfer-2] DEBUG nextflow.file.FilePorter - Local cache found for foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq at /Users/benjaminwehnert/dmscore/work/stage-36458275-f468-409a-bb18-98996facae84/cc/f938a8775ac8befd4337b2732c7dc9/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq -Apr-30 01:54:47.399 [FileTransfer-1] DEBUG nextflow.file.FilePorter - Local cache found for foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq at /Users/benjaminwehnert/dmscore/work/stage-36458275-f468-409a-bb18-98996facae84/28/abb932c7e69700944b1605c1ee72d2/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq -Apr-30 01:54:47.445 [Actor Thread 4] INFO nextflow.processor.TaskProcessor - [9e/d2b28e] Cached process > NFCORE_DMSCORE:DMSCORE:FASTQC (gid1a_1_quality_1_pe) -Apr-30 01:54:47.483 [Actor Thread 4] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'PublishDir' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false -Apr-30 01:54:47.515 [Actor Thread 5] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 2; slices: 1; internal sort time: 0.002 s; external sort time: 0.001 s; total time: 0.003 s -Apr-30 01:54:47.515 [Actor Thread 5] DEBUG nextflow.file.FileCollector - >> temp file exists? true -Apr-30 01:54:47.517 [Actor Thread 5] DEBUG nextflow.file.FileCollector - Retrieved cached collect-files from: /Users/benjaminwehnert/dmscore/work/collect-file/0dde6dd9bfe7393330209153ab0f204f -Apr-30 01:54:47.520 [Actor Thread 5] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-1489236878513704241 -Apr-30 01:54:51.271 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 1; name: NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/63/e5edfa2e469709a5513d0401c69f03] -Apr-30 01:54:51.275 [Task monitor] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'TaskFinalizer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false -Apr-30 01:54:51.321 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-30 01:54:51.326 [Task submitter] INFO nextflow.Session - [2b/9ebde1] Submitted process > NFCORE_DMSCORE:DMSCORE:MULTIQC -Apr-30 01:54:51.410 [Actor Thread 12] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process -Apr-30 01:55:01.965 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 3; name: NFCORE_DMSCORE:DMSCORE:MULTIQC; status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/2b/9ebde1b8d8ace30edde0fba5d024f1] -Apr-30 01:55:02.002 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-30 01:55:02.003 [Task submitter] INFO nextflow.Session - [44/fb3934] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_MEM (gid1a_1_quality_1_pe) -Apr-30 01:55:02.148 [TaskFinalizer-2] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:MULTIQC > Skipping output binding because one or more optional files are missing: fileoutparam<2> -Apr-30 01:55:06.427 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 4; name: NFCORE_DMSCORE:DMSCORE:BWA_MEM (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/44/fb39348cda0e426f67e945b3b453c9] -Apr-30 01:55:06.450 [TaskFinalizer-3] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<1:1> -Apr-30 01:55:06.450 [TaskFinalizer-3] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<2:1> -Apr-30 01:55:06.450 [TaskFinalizer-3] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<3:1> -Apr-30 01:55:06.503 [Actor Thread 4] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process -Apr-30 01:55:06.516 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-30 01:55:06.517 [Task submitter] INFO nextflow.Session - [4f/32eeb6] Submitted process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS (gid1a_1_quality_1_pe) -Apr-30 01:55:10.625 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 5; name: NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/4f/32eeb651f29b281170859a5ccea65d] -Apr-30 01:55:10.698 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-30 01:55:10.698 [Task submitter] INFO nextflow.Session - [1f/f5a995] Submitted process > NFCORE_DMSCORE:DMSCORE:PREMERGE (gid1a_1_quality_1_pe) -Apr-30 01:55:13.152 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 6; name: NFCORE_DMSCORE:DMSCORE:PREMERGE (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/1f/f5a99582c50d984c21bf1d7f2ff192] -Apr-30 01:55:13.178 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -Apr-30 01:55:13.178 [Task submitter] INFO nextflow.Session - [97/8e2b73] Submitted process > NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe) -Apr-30 01:58:37.622 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 7; name: NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe); status: COMPLETED; exit: 127; error: -; workDir: /Users/benjaminwehnert/dmscore/work/97/8e2b7342dc0c54a5e5720f3f01e2e9] -Apr-30 01:58:37.686 [TaskFinalizer-6] DEBUG nextflow.processor.TaskProcessor - Handling unexpected condition for - task: name=NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe); work-dir=/Users/benjaminwehnert/dmscore/work/97/8e2b7342dc0c54a5e5720f3f01e2e9 - error [nextflow.exception.ProcessFailedException]: Process `NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe)` terminated with an error exit status (127) -Apr-30 01:58:37.742 [TaskFinalizer-6] ERROR nextflow.processor.TaskProcessor - Error executing process > 'NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe)' - -Caused by: - Process `NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe)` terminated with an error exit status (127) - - -Command executed: - - # Index reference - samtools faidx GID1A_SUNi_ref_small.fasta - gatk CreateSequenceDictionary -R GID1A_SUNi_ref_small.fasta - - # Read start and stop codon from input - start_stop_codon="352-1383" - - # Run GATK AnalyzeSaturationMutagenesis - gatk AnalyzeSaturationMutagenesis -I merged_reads.bam -R GID1A_SUNi_ref_small.fasta --orf $start_stop_codon --paired-mode false --min-q 30 --min-variant-obs 3 -O gatk_output - - # Save versions - cat <<-END_VERSIONS > versions.yml - "NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS": - samtools: $(samtools --version |& sed '1!d ; s/samtools //') - gatk: $(gatk --version |& sed 's/^.*GATK//') - END_VERSIONS - -Command exit status: - 127 - -Command output: - (empty) - -Command error: - Unable to find image 'community.wave.seqera.io/library/gatk_samtools_java-1.7.0-openjdk-conda-aarch64:50bbe1742b62bc8f' locally - 50bbe1742b62bc8f: Pulling from library/gatk_samtools_java-1.7.0-openjdk-conda-aarch64 - b9cdf96edaf9: Pulling fs layer - 3e5b316ff8d6: Pulling fs layer - b9cdf96edaf9: Download complete - 3e5b316ff8d6: Download complete - Digest: sha256:ab1b440a72d9caec2beb4d205294eb7d43c33448ccb863597ae6858674718d69 - Status: Downloaded newer image for community.wave.seqera.io/library/gatk_samtools_java-1.7.0-openjdk-conda-aarch64:50bbe1742b62bc8f - .command.sh: line 10: gatk: command not found - -Work dir: - /Users/benjaminwehnert/dmscore/work/97/8e2b7342dc0c54a5e5720f3f01e2e9 - -Tip: when you have fixed the problem you can continue the execution adding the option `-resume` to the run command line -Apr-30 01:58:37.755 [main] DEBUG nextflow.Session - Session await > all processes finished -Apr-30 01:58:37.779 [TaskFinalizer-6] INFO nextflow.Session - Execution cancelled -- Finishing pending tasks before exit -Apr-30 01:58:37.797 [Task monitor] DEBUG n.processor.TaskPollingMonitor - <<< barrier arrives (monitor: local) - terminating tasks monitor poll loop -Apr-30 01:58:37.822 [main] DEBUG nextflow.Session - Session await > all barriers passed -Apr-30 01:58:37.883 [TaskFinalizer-6] ERROR nextflow.Nextflow - Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting -Apr-30 01:58:37.884 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'TaskFinalizer' shutdown completed (hard=false) -Apr-30 01:58:37.886 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'PublishDir' shutdown completed (hard=false) -Apr-30 01:58:37.900 [main] INFO nextflow.Nextflow - -[nf-core/dmscore] Pipeline completed with errors- -Apr-30 01:58:37.908 [main] DEBUG n.trace.WorkflowStatsObserver - Workflow completed > WorkflowStats[succeededCount=5; failedCount=1; ignoredCount=0; cachedCount=1; pendingCount=0; submittedCount=0; runningCount=0; retriesCount=0; abortedCount=0; succeedDuration=24.5s; failedDuration=13m 37s; cachedDuration=48s;loadCpus=0; loadMemory=0; peakRunning=2; peakCpus=5; peakMemory=14 GB; ] -Apr-30 01:58:37.909 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow completed -- saving trace file -Apr-30 01:58:37.912 [main] DEBUG nextflow.trace.ReportObserver - Workflow completed -- rendering execution report -Apr-30 01:58:39.474 [main] DEBUG nextflow.trace.TimelineObserver - Workflow completed -- rendering execution timeline -Apr-30 01:58:39.678 [main] DEBUG nextflow.cache.CacheDB - Closing CacheDB done -Apr-30 01:58:39.701 [main] INFO org.pf4j.AbstractPluginManager - Stop plugin 'nf-schema@2.3.0' -Apr-30 01:58:39.701 [main] DEBUG nextflow.plugin.BasePlugin - Plugin stopped nf-schema -Apr-30 01:58:39.702 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'FileTransfer' shutdown completed (hard=false) -Apr-30 01:58:39.704 [main] DEBUG nextflow.script.ScriptRunner - > Execution complete -- Goodbye diff --git a/modules/local/dmsanalysis/bin/SeqDepth_simulation.R b/modules/local/dmsanalysis/bin/SeqDepth_simulation.R new file mode 100644 index 0000000..2207429 --- /dev/null +++ b/modules/local/dmsanalysis/bin/SeqDepth_simulation.R @@ -0,0 +1,131 @@ +# input: prefiltered (by codon library) gatk path, possible mutations path, output_folder, reduction_fraction (steps in % to reduce counts), threshold to count variant as present in dataset +# output: pdf showing the plot +# limitation: takes quite a lot time: depends on number of counts in prefiltered gatk (4 min on M1 MacBook for 340,000 counts in total) -> reduction_fraction only has a low impact -> need to find more efficient random-sampling algorithm + +library(dplyr) +library(ggplot2) + +SeqDepth_simulation_plot <- function(prefiltered_gatk_path, possible_mutations_path, output_file_path, reduction_fraction = 0.01, threshold = 3) { + + # Read data from the specified CSV file + data <- read.csv(prefiltered_gatk_path) + data <- data %>% mutate(counts = as.numeric(counts)) # Ensure counts are numeric + original_counts <- data$counts # Store the original counts for weight calculations + possible_mutations <- read.csv(possible_mutations_path) + + # Initialize variables + total_counts <- sum(data$counts) + reduction_per_step <- floor(total_counts * reduction_fraction) # Round down to the nearest integer + results <- data.frame(remaining_counts = numeric(), remaining_variants = numeric()) + + # Track the initial state + remaining_variants <- sum(data$counts >= threshold) + remaining_counts <- total_counts + results <- rbind(results, data.frame(remaining_counts = remaining_counts, remaining_variants = remaining_variants)) + + # Get a list of indices for non-zero variants + non_zero_indices <- which(data$counts > 0) + + # Calculate initial weights based on the original counts for non-zero variants + weights <- as.numeric(original_counts[non_zero_indices]) + weights <- weights / sum(weights) # Normalize weights + + # Loop until all counts are zero + while (remaining_counts > 0) { + # If no more non-zero variants are available, break the loop + if (length(non_zero_indices) == 0) { + break + } + + # Randomly reduce counts by the specified amount using weighted sampling + for (i in 1:reduction_per_step) { + # Randomly choose an index from the non-zero variants based on weights + selected_idx <- sample(length(non_zero_indices), 1, prob = weights) + index <- non_zero_indices[selected_idx] + + # Reduce the count by 1 + data$counts[index] <- data$counts[index] - 1 + + # If the count reaches zero, remove the index from the list of non-zero variants + if (data$counts[index] == 0) { + non_zero_indices <- non_zero_indices[-selected_idx] + weights <- weights[-selected_idx] # Remove the corresponding weight + } + } + + # Update remaining counts and remaining variants after the reduction + remaining_counts <- sum(data$counts) + remaining_variants <- sum(data$counts >= threshold & data$counts > 0) + + # Store the results for this step + results <- rbind(results, data.frame(remaining_counts = remaining_counts, remaining_variants = remaining_variants)) + + # Adjust the reduction_per_step if the total remaining counts are less than the reduction amount + if (remaining_counts < reduction_per_step) { + reduction_per_step <- remaining_counts + } + + # Recalculate weights using the original counts, but only for remaining non-zero variants + if (length(non_zero_indices) > 0) { + weights <- as.numeric(original_counts[non_zero_indices]) + weights <- weights / sum(weights) # Normalize weights + } + } + + # Transform results for plotting + baseline_count <- max(results$remaining_counts) + plot_data <- results %>% + mutate( + remaining_counts_fold = round(remaining_counts / baseline_count, 2), # X-axis in fold-change from max, rounded to 2 decimals + remaining_variants_percent = (remaining_variants / nrow(possible_mutations)) * 100 # Y-axis in percent + ) + + # Set plot limits + x_max <- 1 # X-axis limit exactly at 1 + y_max <- 100 # Y-axis limit at 100% + + # Create the plot + plot <- ggplot(plot_data, aes(x = remaining_counts_fold, y = remaining_variants_percent)) + + geom_line(color = "black", size = 0.4) + # Solid line for the data + geom_hline(yintercept = 100, linetype = "dotted", color = "black") + # Horizontal line at 100% + + # Main plot settings with fine grid lines + scale_y_continuous( + labels = scales::percent_format(scale = 1), + limits = c(0, y_max), + breaks = seq(0, y_max, by = 5) # Y-axis grid lines every 5% + ) + + scale_x_continuous( + limits = c(0, x_max), + breaks = seq(0, x_max, length.out = 20), # X-axis with 20 grid lines ending at 1 + labels = scales::number_format(accuracy = 0.01) # Round labels to 2 decimal places + ) + + labs( + x = "Fold-Change of Sequencing Depth", + y = "Variants (% of Maximum)" + ) + + theme_minimal() + + theme( + panel.border = element_rect(color = "black", fill = NA), # Add a box-like border + panel.grid.major = element_line(size = 0.2, linetype = "solid", color = "grey80"), # Fine grid lines + panel.grid.minor = element_blank(), # No minor grid lines for clarity + axis.text.x = element_text(angle = 45, hjust = 1) # Rotate X-axis labels at 45 degrees + ) + + # Save the plot as a PDF in the specified output folder + ggsave(output_file_path, plot = plot, device = "pdf", width = 8, height = 6) +} + +#Example usage with input and output paths +# results_weighted <- coverage_simulation_plot( +# prefiltered_gatk_path = "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library.csv", +# possible_mutations_path = "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/possible_mutations.csv", +# output_folder = "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs", +# reduction_fraction = 0.01, +# threshold = 3 +# ) + + +#SeqDepth + + diff --git a/modules/local/dmsanalysis/bin/aa_seq.R b/modules/local/dmsanalysis/bin/aa_seq.R new file mode 100644 index 0000000..9a39fb5 --- /dev/null +++ b/modules/local/dmsanalysis/bin/aa_seq.R @@ -0,0 +1,37 @@ +# input: wildtype-seq, start&stopp pos. +# output: amino acid sequence within the start-stop frame (.txt) + +# Load necessary libraries +suppressMessages(library(Biostrings)) + +# Define the function +aa_seq <- function(wt_seq_input, pos_range, output_file) { + # Parse the start and stop positions from the input format "start-stop" + positions <- unlist(strsplit(pos_range, "-")) + start_pos <- as.numeric(positions[1]) + stop_pos <- as.numeric(positions[2]) + + # Check if the input is a file or a string + if (file.exists(wt_seq_input)) { + # If it's a file, read the sequence from the fasta file + seq_data <- readDNAStringSet(filepath = wt_seq_input) + wt_seq <- seq_data[[1]] # Extract the sequence + } else { + # Otherwise, treat the input as a sequence string + wt_seq <- DNAString(wt_seq_input) + } + + # Extract the sequence between start and stop codons + coding_seq <- subseq(wt_seq, start = start_pos, end = stop_pos) + + # Translate the coding sequence into an amino acid sequence + aa_seq <- translate(coding_seq) + + # Write the amino acid sequence to a .txt file + write(as.character(aa_seq), file = output_file) +} + +# Example usage: +# translate_to_protein("/path/to/sequence.fasta", "23-1225", "/path/to/output_protein.txt") + +#aa_seq("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/MORtn5_reference.fa", "23-1225", "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/aa_seq.txt") diff --git a/modules/local/dmsanalysis/bin/complete_prefiltered_gatk.R b/modules/local/dmsanalysis/bin/complete_prefiltered_gatk.R new file mode 100644 index 0000000..1e16adc --- /dev/null +++ b/modules/local/dmsanalysis/bin/complete_prefiltered_gatk.R @@ -0,0 +1,79 @@ +# input: NNK_codon_library_filtered_gatk.csv-path, prefiltered_gatk.csv-path (containing only NNK mutations), output_folder-path +# output: completed gatk_file with all possible variants (even if not measured in sequencing) -> NA in counts and counts_per_cov to 0.0000001 to deal with log-scale in following calculations + +library(dplyr) +library(Biostrings) # Required for codon-to-amino-acid translation + +# Function to calculate Hamming distance (varying_bases) +hamming_distance <- function(wt_codon, variant_codon) { + sum(strsplit(wt_codon, "")[[1]] != strsplit(variant_codon, "")[[1]]) +} + +# Function to get amino acid from codon +get_amino_acid <- function(codon) { + codon_table <- GENETIC_CODE + aa <- codon_table[[toupper(codon)]] + if (is.null(aa)) { + return(NA) # Handle cases where codon is not valid + } + return(aa) +} + +# Function to calculate mutation type (aa_mut) and pos_mut +mutation_details <- function(wt_codon, variant_codon, codon_number) { + wt_aa <- get_amino_acid(wt_codon) + variant_aa <- get_amino_acid(variant_codon) + + # If amino acids are different, it's a missense mutation; otherwise, synonymous + if (wt_aa != variant_aa) { + mutation_type <- "M" # Missense mutation + } else { + mutation_type <- "S" # Synonymous mutation + } + + # aa_mut: Type of mutation and amino acid changes (e.g., M:D>S) + aa_mut <- paste0(mutation_type, ":", wt_aa, ">", variant_aa) + + # pos_mut: Wild-type AA, codon position, mutated AA (e.g., D2Q) + pos_mut <- paste0(wt_aa, codon_number, variant_aa) + + return(list(aa_mut = aa_mut, pos_mut = pos_mut)) +} + +complete_prefiltered_gatk <- function(possible_nnk_path, prefiltered_gatk_path, output_file_path) { + + # Load the possible NNK mutations CSV + possible_nnk <- read.csv(possible_nnk_path) + + # Load the prefiltered GATK CSV + prefiltered_gatk <- read.csv(prefiltered_gatk_path) + + # Create codon_mut column in possible_NNK_mutations in the format 'Codon_Number:wt_codon>Variant' + possible_nnk <- possible_nnk %>% + mutate(codon_mut = paste0(Codon_Number, ":", wt_codon, ">", Variant)) + + # Merge both dataframes based on the codon_mut column (full join to include all) + merged_data <- full_join(prefiltered_gatk, possible_nnk, by = "codon_mut") + + # Fill missing values in counts_per_cov and counts with 0.0000001 + merged_data <- merged_data %>% + mutate(counts_per_cov = ifelse(is.na(counts_per_cov), 0.0000001, counts_per_cov), + counts = ifelse(is.na(counts), 0.000001, counts)) + + # Calculate Hamming distance (varying_bases) and mutation details (aa_mut, pos_mut) + merged_data <- merged_data %>% + rowwise() %>% + mutate(varying_bases = hamming_distance(wt_codon, Variant), + mutation_info = list(mutation_details(wt_codon, Variant, Codon_Number))) %>% + mutate(aa_mut = mutation_info$aa_mut, # Extract aa_mut + pos_mut = mutation_info$pos_mut) %>% # Extract pos_mut + ungroup() %>% + select(-mutation_info) # Remove the temporary list column + + # Save the merged data to a new CSV file + write.csv(merged_data, file = output_file_path, row.names = FALSE) +} + +# Example call +#complete_prefiltered_gatk("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/possible_NNK_mutations.csv", "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library.csv", "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs") + diff --git a/modules/local/dmsanalysis/bin/counts_heatmap.R b/modules/local/dmsanalysis/bin/counts_heatmap.R new file mode 100644 index 0000000..dd37b2e --- /dev/null +++ b/modules/local/dmsanalysis/bin/counts_heatmap.R @@ -0,0 +1,154 @@ +# Input: prepared GATK data path, output_path, threshold (same as used for prepare_gatk_data_for_counts_per_cov_heatmap function) +# Output: counts_per_cov_heatmap.pdf + +library(dplyr) +library(ggplot2) + +counts_heatmap <- function(input_csv_path, threshold = 3, output_pdf_path, img_format = "pdf") { + + # Inner function to add padding to the last row, adding 21 amino acids per position + pad_heatmap_data_long <- function(heatmap_data_long, min_non_na_value, num_positions_per_row = 75) { + all_amino_acids <- c("A", "C", "D", "E", "F", "G", "H", "I", "K", "L", + "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y", "*") + + max_position <- max(heatmap_data_long$position) + num_missing_positions <- num_positions_per_row - (max_position %% num_positions_per_row) + + if (num_missing_positions < num_positions_per_row) { + new_positions <- (max_position + 1):(max_position + num_missing_positions) + + # Add all 21 amino acid variants for each new position + padding_data <- expand.grid( + mut_aa = all_amino_acids, # All possible amino acids + position = new_positions # New positions to be padded + ) + + # Set placeholder values for the added positions to the exact smallest non-NA value + padding_data$total_counts <- min_non_na_value # Set to the smallest non-NA value + padding_data$wt_aa <- "Y" # Set wild-type amino acid to 'Y' + padding_data$wt_aa_pos <- paste0("Y", padding_data$position) # Create wt_aa_pos with correct positions + padding_data$row_group <- max(heatmap_data_long$row_group) # Set row group to the current last group + + # Add the new padding rows to heatmap_data_long + heatmap_data_long <- dplyr::bind_rows(heatmap_data_long, padding_data) + } + + return(heatmap_data_long) + } + + # Load the CSV data + heatmap_data <- read.csv(input_csv_path) + + # Check if the necessary column exists in the data + if (!"total_counts" %in% colnames(heatmap_data)) { + stop("The column 'total_counts' is not found in the data.") + } + + # Create heatmap_data_long by selecting necessary columns + heatmap_data_long <- heatmap_data %>% + select(mut_aa, position, total_counts, wt_aa) # Use 'total_counts' + + # Find the smallest non-NA value in total_counts + min_non_na_value <- min(heatmap_data_long$total_counts, na.rm = TRUE) + + # Group positions by rows (75 positions per row) and calculate row_group + heatmap_data_long <- heatmap_data_long %>% + mutate(row_group = ((position - 1) %/% 75) + 1) # Grouping positions into rows + + # Apply padding to add missing positions at the end of the last row, using the calculated min value + heatmap_data_long <- pad_heatmap_data_long(heatmap_data_long, min_non_na_value) + + # Convert positions to numeric, sort them, and create wt_aa_pos for the plot + heatmap_data_long <- heatmap_data_long %>% + mutate(position = as.numeric(position)) %>% # Ensure position is numeric + arrange(position) %>% # Sort by position + mutate(wt_aa_pos = factor(paste0(wt_aa, position), levels = unique(paste0(wt_aa, position)))) # Create sorted factor levels for wt_aa_pos + + # Add a column to identify synonymous mutations (where mut_aa == wt_aa) + heatmap_data_long <- heatmap_data_long %>% + mutate(synonymous = mut_aa == wt_aa) + + # Definiere die korrekte Reihenfolge der Aminosäuren + amino_acid_order <- c("*", "A", "C", "D", "E", "F", "G", "H", "I", "K", + "L", "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y") + + # Bearbeite heatmap_data_long und erstelle syn_positions gleichzeitig + syn_positions <- heatmap_data_long %>% + mutate(mut_aa = factor(mut_aa, levels = amino_acid_order), + # Berechne die x-Koordinate, die pro Gruppe immer von 1 bis 75 verläuft + x = as.numeric(factor(wt_aa_pos, levels = unique(wt_aa_pos))) - ((row_group - 1) * 75), + y = as.numeric(factor(mut_aa, levels = amino_acid_order))) %>% + filter(synonymous == TRUE) + + # Calculate the number of row groups and adjust plot height dynamically + num_row_groups <- max(heatmap_data_long$row_group) + plot_height <- num_row_groups * 4 + + # Set the limits for the color scale, ignoring NA (negative values are now NA) + min_count <- min(heatmap_data_long$total_counts, na.rm = TRUE) + max_count <- max(heatmap_data_long$total_counts, na.rm = TRUE) + max_position <- max(heatmap_data$position) + + # Create the heatmap plot with explicit handling for positions > max_position + heatmap_plot <- ggplot(heatmap_data_long, aes(x = wt_aa_pos, y = mut_aa, fill = total_counts)) + + scale_fill_gradientn(colours = c(alpha("blue", 0), "blue"), na.value = "grey35", trans = "log", # Apply log transformation to the scale + limits = c(min_count, max_count), + breaks = scales::trans_breaks("log10", function(x) 10^x), # Logarithmic scale breaks + labels = scales::trans_format("log10", scales::math_format(10^.x))) + + scale_x_discrete(labels = function(x) { + numeric_pos <- as.numeric(gsub("[^0-9]", "", x)) + ifelse(numeric_pos > max_position, " ", x) + }) + + geom_tile() + + + # Add diagonal lines for synonymous mutations using geom_segment + geom_segment(data = syn_positions[syn_positions$position <= max_position, ], + aes(x = x - 0.485, xend = x + 0.485, + y = y - 0.485, yend = y + 0.485, color = synonymous), + size = 0.2) + + + # Manuelle Farbskala für die diagonalen Linien + scale_color_manual(values = c("TRUE" = "grey10"), labels = c("TRUE" = "")) + + + theme_minimal() + + labs(title = "Heatmap of Counts per Variant", x = "Wild-type Amino Acid", y = "Mutant Amino Acid", fill = "Counts", color = "Synonymous Mutation") + + theme(plot.title = element_text(size = 16, face = "bold"), + axis.text.x = element_text(angle = 90, hjust = 1, vjust = 0.5, size = 12), + axis.text.y = element_text(size = 12), # Larger y-axis labels + axis.title.x = element_text(size = 16), + axis.title.y = element_text(size = 16), + legend.title = element_text(size = 14), # Larger legend title + legend.text = element_text(size = 12), # Larger legend text + panel.spacing = unit(0.1, "lines"), # Adjust panel spacing + strip.text = element_blank(), # Remove row group labels (facet numbers) + strip.background = element_blank(), + panel.grid.major = element_blank(), # Remove major grid lines + panel.grid.minor = element_blank()) + # Remove minor grid lines + facet_wrap(~ row_group, scales = "free_x", ncol = 1) + # Group by 75 positions per row + theme(panel.spacing = unit(0.2, "lines")) + + heatmap_plot <- heatmap_plot + + geom_point(data = heatmap_data_long, aes(size = ""), colour = "black", alpha = 0) # Invisible points for legend + heatmap_plot <- heatmap_plot + + guides(size = guide_legend(paste("Dropout (Counts <", threshold, ")"), override.aes = list(shape = 15, size = 8, colour = "grey35", alpha = 1))) # Define Legend for Dropouts + + # Save the heatmap plot + if (img_format == "pdf") { + ggsave(output_pdf_path, plot = heatmap_plot, width = 16, height = plot_height, dpi = 150, device = cairo_pdf) + } else { + ggsave(output_pdf_path, plot = heatmap_plot, width = 16, height = plot_height, dpi = 150) + } + + if (file.exists(output_pdf_path)) { + print("Heatmap image successfully created!") + } else { + print("Error: Heatmap image was not created.") + } +} + +# Test the function +# counts_heatmap( +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/prepared_gatk_data.csv", +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/counts_heatmap.pdf", +# threshold = 3 +# ) diff --git a/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R b/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R new file mode 100644 index 0000000..8af6b80 --- /dev/null +++ b/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R @@ -0,0 +1,154 @@ +# Input: prefiltered GATK data path, output_path, threshold (same as used for prepare_gatk_data_for_counts_per_cov_heatmap function) +# Output: counts_per_cov_heatmap.pdf + +library(dplyr) +library(ggplot2) + +counts_per_cov_heatmap <- function(input_csv_path, threshold = 3, output_pdf_path, img_format = "pdf") { + + # Inner function to add padding to the last row, adding 21 amino acids per position + pad_heatmap_data_long <- function(heatmap_data_long, min_non_na_value, num_positions_per_row = 75) { + all_amino_acids <- c("A", "C", "D", "E", "F", "G", "H", "I", "K", "L", + "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y", "*") + + max_position <- max(heatmap_data_long$position) + num_missing_positions <- num_positions_per_row - (max_position %% num_positions_per_row) + + if (num_missing_positions < num_positions_per_row) { + new_positions <- (max_position + 1):(max_position + num_missing_positions) + + # Add all 21 amino acid variants for each new position + padding_data <- expand.grid( + mut_aa = all_amino_acids, # All possible amino acids + position = new_positions # New positions to be padded + ) + + # Set placeholder values for the added positions to the exact smallest non-NA value + padding_data$total_counts_per_cov <- min_non_na_value # Set to the smallest non-NA value + padding_data$wt_aa <- "Y" # Set wild-type amino acid to 'Y' + padding_data$wt_aa_pos <- paste0("Y", padding_data$position) # Create wt_aa_pos with correct positions + padding_data$row_group <- max(heatmap_data_long$row_group) # Set row group to the current last group + + # Add the new padding rows to heatmap_data_long + heatmap_data_long <- dplyr::bind_rows(heatmap_data_long, padding_data) + } + + return(heatmap_data_long) + } + + # Load the CSV data + heatmap_data <- read.csv(input_csv_path) + + # Check if the necessary column exists in the data + if (!"total_counts_per_cov" %in% colnames(heatmap_data)) { + stop("The column 'total_counts_per_cov' is not found in the data.") + } + + # Create heatmap_data_long by selecting necessary columns + heatmap_data_long <- heatmap_data %>% + select(mut_aa, position, total_counts_per_cov, wt_aa) # Use 'total_counts_per_cov' + + # Find the smallest non-NA value in total_counts_per_cov + min_non_na_value <- min(heatmap_data_long$total_counts_per_cov, na.rm = TRUE) + + # Group positions by rows (75 positions per row) and calculate row_group + heatmap_data_long <- heatmap_data_long %>% + mutate(row_group = ((position - 1) %/% 75) + 1) # Grouping positions into rows + + # Apply padding to add missing positions at the end of the last row, using the calculated min value + heatmap_data_long <- pad_heatmap_data_long(heatmap_data_long, min_non_na_value) + + # Convert positions to numeric, sort them, and create wt_aa_pos for the plot + heatmap_data_long <- heatmap_data_long %>% + mutate(position = as.numeric(position)) %>% # Ensure position is numeric + arrange(position) %>% # Sort by position + mutate(wt_aa_pos = factor(paste0(wt_aa, position), levels = unique(paste0(wt_aa, position)))) # Create sorted factor levels for wt_aa_pos + + # Add a column to identify synonymous mutations (where mut_aa == wt_aa) + heatmap_data_long <- heatmap_data_long %>% + mutate(synonymous = mut_aa == wt_aa) + + # Definiere die korrekte Reihenfolge der Aminosäuren + amino_acid_order <- c("*", "A", "C", "D", "E", "F", "G", "H", "I", "K", + "L", "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y") + + # Bearbeite heatmap_data_long und erstelle syn_positions gleichzeitig + syn_positions <- heatmap_data_long %>% + mutate(mut_aa = factor(mut_aa, levels = amino_acid_order), + # Berechne die x-Koordinate, die pro Gruppe immer von 1 bis 75 verläuft + x = as.numeric(factor(wt_aa_pos, levels = unique(wt_aa_pos))) - ((row_group - 1) * 75), + y = as.numeric(factor(mut_aa, levels = amino_acid_order))) %>% + filter(synonymous == TRUE) + + # Calculate the number of row groups and adjust plot height dynamically + num_row_groups <- max(heatmap_data_long$row_group) + plot_height <- num_row_groups * 4 + + # Set the limits for the color scale, ignoring NA (negative values are now NA) + min_count <- min(heatmap_data_long$total_counts_per_cov, na.rm = TRUE) + max_count <- max(heatmap_data_long$total_counts_per_cov, na.rm = TRUE) + max_position <- max(heatmap_data$position) + + # Create the heatmap plot with explicit handling for positions > max_position + heatmap_plot <- ggplot(heatmap_data_long, aes(x = wt_aa_pos, y = mut_aa, fill = total_counts_per_cov)) + + scale_fill_gradientn(colours = c(alpha("blue", 0), "blue"), na.value = "grey35", trans = "log", # Apply log transformation to the scale + limits = c(min_count, max_count), + breaks = scales::trans_breaks("log10", function(x) 10^x), # Logarithmic scale breaks + labels = scales::trans_format("log10", scales::math_format(10^.x))) + + scale_x_discrete(labels = function(x) { + numeric_pos <- as.numeric(gsub("[^0-9]", "", x)) + ifelse(numeric_pos > max_position, " ", x) + }) + + geom_tile() + + + # Add diagonal lines for synonymous mutations using geom_segment + geom_segment(data = syn_positions[syn_positions$position <= max_position, ], + aes(x = x - 0.485, xend = x + 0.485, + y = y - 0.485, yend = y + 0.485, color = synonymous), + size = 0.2) + + + # Manuelle Farbskala für die diagonalen Linien + scale_color_manual(values = c("TRUE" = "grey10"), labels = c("TRUE" = "")) + + + theme_minimal() + + labs(title = "Heatmap of Counts per Coverage for Mutations", x = "Wild-type Amino Acid", y = "Mutant Amino Acid", fill = "Counts per \n Coverage", color = "Synonymous Mutation") + + theme(plot.title = element_text(size = 16, face = "bold"), + axis.text.x = element_text(angle = 90, hjust = 1, vjust = 0.5, size = 12), + axis.text.y = element_text(size = 12), # Larger y-axis labels + axis.title.x = element_text(size = 16), + axis.title.y = element_text(size = 16), + legend.title = element_text(size = 14), # Larger legend title + legend.text = element_text(size = 12), # Larger legend text + panel.spacing = unit(0.1, "lines"), # Adjust panel spacing + strip.text = element_blank(), # Remove row group labels (facet numbers) + strip.background = element_blank(), + panel.grid.major = element_blank(), # Remove major grid lines + panel.grid.minor = element_blank()) + # Remove minor grid lines + facet_wrap(~ row_group, scales = "free_x", ncol = 1) + # Group by 75 positions per row + theme(panel.spacing = unit(0.2, "lines")) + + heatmap_plot <- heatmap_plot + + geom_point(data = heatmap_data_long, aes(size = ""), colour = "black", alpha = 0) # Invisible points for legend + heatmap_plot <- heatmap_plot + + guides(size = guide_legend(paste("Dropout (Counts <", threshold, ")"), override.aes = list(shape = 15, size = 8, colour = "grey35", alpha = 1))) # Define Legend for Dropouts + + # Save the heatmap plot + if (img_format == "pdf") { + ggsave(output_pdf_path, plot = heatmap_plot, width = 16, height = plot_height, dpi = 150, device = cairo_pdf) + } else { + ggsave(output_pdf_path, plot = heatmap_plot, width = 16, height = plot_height, dpi = 150) + } + + if (file.exists(output_pdf_path)) { + print("Heatmap image successfully created!") + } else { + print("Error: Heatmap image was not created.") + } +} + +# Test the function +# counts_per_cov_heatmap( +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/prepared_gatk_data.csv", +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/heatmap.pdf", +# threshold = 3 +# ) diff --git a/modules/local/dmsanalysis/bin/detect_codons.R b/modules/local/dmsanalysis/bin/detect_codons.R new file mode 100644 index 0000000..0c35b3b --- /dev/null +++ b/modules/local/dmsanalysis/bin/detect_codons.R @@ -0,0 +1,65 @@ +# input: wildtype nucleotide sequence as string or .fa +# output: first nucleotide position of start codon and last nucleotide position of stopp codon (e.g. "7-2145") + +# Load necessary package +if (!requireNamespace("BiocManager", quietly = TRUE)) { + install.packages("BiocManager") +} +if (!requireNamespace("Biostrings", quietly = TRUE)) { + BiocManager::install("Biostrings") +} +suppressMessages(library(Biostrings)) + +# Function to find the nearest stop codon in the reading frame of the first start codon +# Accepts either a sequence string or a path to a reference FASTA file +find_start_stop <- function(input_string, start_codon = "ATG", stop_codons = c("TAA", "TAG", "TGA")) { + # Determine if the input is a file path or a raw sequence + if (file.exists(input_string)) { + # If the input is a valid file path, read the DNA sequence from the FASTA file + dna <- readDNAStringSet(input_string) + sequence <- as.character(dna[[1]]) # Extract the sequence as a character string + } else { + # If the input is not a file path, treat it as a raw sequence + sequence <- input_string + } + + # Check if the sequence is provided or successfully read + if (is.null(sequence)) { + stop("No valid sequence or FASTA file provided.") + } + + # Convert the sequence to uppercase to ensure matching regardless of case + sequence <- toupper(sequence) + + # Function to find codons in the sequence + find_codons <- function(sequence, codon) { + codon_positions <- gregexpr(codon, sequence)[[1]] + codon_positions[codon_positions > 0] # Return only positive matches + } + + # Search for the first start codon + start_positions <- find_codons(sequence, start_codon) + + if (length(start_positions) == 0 || start_positions[1] == -1) { + return("No start codon found") # Return message if no start codon is found + } + first_start <- start_positions[1] # Consider only the first start codon + + # Search for all possible stop codons + stop_positions_list <- lapply(stop_codons, find_codons, sequence = sequence) + all_stop_positions <- sort(unlist(stop_positions_list)) + + # Filter only stop codons that are in the same reading frame and come after the start codon + in_frame_stops <- all_stop_positions[(all_stop_positions - first_start) %% 3 == 0 & all_stop_positions > first_start] + if (length(in_frame_stops) > 0) { + # Choose the nearest stop codon + closest_stop <- min(in_frame_stops) + return(cat(paste(first_start, closest_stop + 2, sep = "-"))) # Output format: start-stop + } else { + return("No suitable stop codon found") # If no suitable stop codon is found + } +} + +#find_start_stop("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/MORtn5_reference.fa") + + diff --git a/modules/local/dmsanalysis/bin/filter_gatk_by_codon_library.R b/modules/local/dmsanalysis/bin/filter_gatk_by_codon_library.R new file mode 100644 index 0000000..15ee22d --- /dev/null +++ b/modules/local/dmsanalysis/bin/filter_gatk_by_codon_library.R @@ -0,0 +1,124 @@ +# Input: pre_processed_raw_gatk_path, mutation library .csv path (former "possible_NNK_mutations.csv"), output_path +# Output: gatk table filtered for only single-codon mutations that are part of the library + +# Load necessary library +library("dplyr") +library("stringr") + +# filter_gatk_by_codon_library <- function(gatk_file_path, codon_library, output_file_path) { +# # Load the GATK table from the provided file path +# gatk_table <- read.csv(gatk_file_path) +# +# # Predefined codon libraries +# nnk_codons <- c('AAG', 'AAT', 'ATG', 'ATT', 'AGG', 'AGT', 'ACG', 'ACT', +# 'TAG', 'TAT', 'TTG', 'TTT', 'TGG', 'TGT', 'TCG', 'TCT', +# 'GAG', 'GAT', 'GTG', 'GTT', 'GGG', 'GGT', 'GCG', 'GCT', +# 'CAG', 'CAT', 'CTG', 'CTT', 'CGG', 'CGT', 'CCG', 'CCT') +# +# # Check if the codon_library is a predefined string or a custom vector +# if (is.character(codon_library) && length(codon_library) == 1) { +# if (codon_library == "NNK") { +# codon_set <- nnk_codons +# } else { +# stop("Invalid predefined codon library specified.") +# } +# } else if (is.vector(codon_library)) { +# codon_set <- codon_library +# } else { +# stop("Invalid codon library format. Must be a predefined string or a custom vector.") +# } +# +# # Filter for single-codon mutations that are part of the codon library and make sure to handle some mistaken formatting from gatk (there are cases where more than 3 Bases are mutated, but column varying_codons == 1) +# filtered_gatk <- gatk_table %>% +# filter(varying_codons == 1 & sub(".*>", "", codon_mut) %in% codon_set) %>% +# rowwise() %>% +# filter({ +# # Split base_mut into individual mutations +# mutations <- unlist(strsplit(base_mut, ",\\s*")) # Splits by comma and removes extra spaces +# # Extract numeric positions from each mutation string +# positions <- as.numeric(str_extract(mutations, "^[0-9]+")) +# +# # Calculate the distance between the first and last position +# distance <- max(positions, na.rm = TRUE) - min(positions, na.rm = TRUE) +# +# # Keep rows where the distance is <= 2 +# distance <= 2 +# }) %>% +# ungroup() +# +# +# # Write the filtered GATK table to the output file path +# write.csv(filtered_gatk, file = output_file_path, row.names = FALSE) +# } +# +# # Example usage with predefined library (NNK): +# # filtered_gatk <- filter_gatk_by_codon_library(raw_gatk, codon_library = "NNK") +# +# # Example usage with custom library: +# # custom_library <- c('AAG', 'TGT', 'GCT') +# # filtered_gatk <- filter_gatk_by_codon_library(raw_gatk, custom_library = custom_library) +# +# #filter_gatk_by_codon_library("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/raw_gatk.csv", codon_library = "NNK", output_file_path = "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library.csv") + + + + + +library("dplyr") +library("stringr") + +filter_gatk_by_codon_library <- function(gatk_file_path, codon_library_path, output_file_path) { + # Load the GATK table from the provided file path + gatk_table <- read.csv(gatk_file_path) + + # Load the codon library from the provided .csv file + codon_library <- read.csv(codon_library_path) + + # Ensure the codon library has the expected columns + if (!all(c("Codon_Number", "wt_codon", "Variant") %in% colnames(codon_library))) { + stop("Codon library must contain columns 'Codon_Number', 'wt_codon', and 'Variant'.") + } + + # Filter the GATK table + filtered_gatk <- gatk_table %>% + filter(varying_codons == 1) %>% # Keep rows with single-codon mutations + rowwise() %>% + filter({ + # Extract the position and mutated codon + codon_position <- as.numeric(sub(":.*", "", codon_mut)) # Extract position before ':' + mutated_codon <- sub(".*>", "", codon_mut) # Extract codon after '>' + + # Check if the position and codon are valid + is_in_library <- any( + codon_library$Codon_Number == codon_position & + (codon_library$Variant == mutated_codon | # Check Variant column + codon_library$wt_codon == mutated_codon) # Check wt_codon column + ) + is_in_library + }) %>% + ungroup() %>% + # Apply additional filtering based on mutation distances + rowwise() %>% + filter({ + # Split base_mut into individual mutations + mutations <- unlist(strsplit(base_mut, ",\\s*")) # Splits by comma and removes extra spaces + # Extract numeric positions from each mutation string + positions <- as.numeric(str_extract(mutations, "^[0-9]+")) + + # Calculate the distance between the first and last position + distance <- max(positions, na.rm = TRUE) - min(positions, na.rm = TRUE) + + # Keep rows where the distance is <= 2 + distance <= 2 + }) %>% + ungroup() + + # Write the filtered GATK table to the output file path + write.csv(filtered_gatk, file = output_file_path, row.names = FALSE) +} + +# example +#filter_gatk_by_codon_library("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/raw_gatk.csv", codon_library = "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/possible_NNK_mutations.csv", output_file_path = "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library.csv") ### this one's is correct for this data set +#filter_gatk_by_codon_library("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/raw_gatk.csv", codon_library = "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/possible_NNK_mutations_taylors_nnk_and_nns.csv", output_file_path = "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/gatk_filtered_by_complete_codon_library.csv") + + diff --git a/modules/local/dmsanalysis/bin/fitness_heatmap.R b/modules/local/dmsanalysis/bin/fitness_heatmap.R new file mode 100644 index 0000000..09e7d57 --- /dev/null +++ b/modules/local/dmsanalysis/bin/fitness_heatmap.R @@ -0,0 +1,160 @@ +library(dplyr) +library(ggplot2) + +fitness_heatmap <- function(input_csv_path, threshold = 3, output_pdf_path, img_format = "pdf") { + + # Inner function to add padding to the last row, adding 21 amino acids per position + pad_heatmap_data_long <- function(heatmap_data_long, min_non_na_value, num_positions_per_row = 75) { + all_amino_acids <- c("A", "C", "D", "E", "F", "G", "H", "I", "K", "L", + "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y", "*") + + max_position <- max(heatmap_data_long$position) + num_missing_positions <- num_positions_per_row - (max_position %% num_positions_per_row) + + if (num_missing_positions < num_positions_per_row) { + new_positions <- (max_position + 1):(max_position + num_missing_positions) + + # Add all 21 amino acid variants for each new position + padding_data <- expand.grid( + mut_aa = all_amino_acids, # All possible amino acids + position = new_positions # New positions to be padded + ) + + # Set placeholder values for the added positions to the exact smallest non-NA value + padding_data$total_counts_per_cov <- min_non_na_value # Set to the smallest non-NA value + padding_data$wt_aa <- "Y" # Set wild-type amino acid to 'Y' + padding_data$wt_aa_pos <- paste0("Y", padding_data$position) # Create wt_aa_pos with correct positions + padding_data$row_group <- max(heatmap_data_long$row_group) # Set row group to the current last group + + # Add the new padding rows to heatmap_data_long + heatmap_data_long <- dplyr::bind_rows(heatmap_data_long, padding_data) + } + + return(heatmap_data_long) + } + + # Load the CSV data + heatmap_data <- read.csv(input_csv_path) + + # Check if the necessary column exists in the data + if (!"fitness" %in% colnames(heatmap_data)) { + stop("The column 'fitness' is not found in the data.") + } + + # Create heatmap_data_long by selecting necessary columns + heatmap_data_long <- heatmap_data %>% + select(mut_aa, position, fitness, wt_aa) # Use 'fitness' + + # Find the smallest non-NA value in fitness column + min_non_na_value <- min(heatmap_data_long$fitness, na.rm = TRUE) + + # Group positions by rows (75 positions per row) and calculate row_group + heatmap_data_long <- heatmap_data_long %>% + mutate(row_group = ((position - 1) %/% 75) + 1) # Grouping positions into rows + + # Apply padding to add missing positions at the end of the last row, using the calculated min value + heatmap_data_long <- pad_heatmap_data_long(heatmap_data_long, min_non_na_value) + + # Convert positions to numeric, sort them, and create wt_aa_pos for the plot + heatmap_data_long <- heatmap_data_long %>% + mutate(position = as.numeric(position)) %>% # Ensure position is numeric + arrange(position) %>% # Sort by position + mutate(wt_aa_pos = factor(paste0(wt_aa, position), levels = unique(paste0(wt_aa, position)))) # Create sorted factor levels for wt_aa_pos + + # Add a column to identify synonymous mutations (where mut_aa == wt_aa) + heatmap_data_long <- heatmap_data_long %>% + mutate(synonymous = mut_aa == wt_aa) + + # Definiere die korrekte Reihenfolge der Aminosäuren + amino_acid_order <- c("*", "A", "C", "D", "E", "F", "G", "H", "I", "K", + "L", "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y") + + # Bearbeite heatmap_data_long und erstelle syn_positions gleichzeitig + syn_positions <- heatmap_data_long %>% + mutate(mut_aa = factor(mut_aa, levels = amino_acid_order), + # Berechne die x-Koordinate, die pro Gruppe immer von 1 bis 75 verläuft + x = as.numeric(factor(wt_aa_pos, levels = unique(wt_aa_pos))) - ((row_group - 1) * 75), + y = as.numeric(factor(mut_aa, levels = amino_acid_order))) %>% + filter(synonymous == TRUE) + + # Calculate the number of row groups and adjust plot height dynamically + num_row_groups <- max(heatmap_data_long$row_group) + plot_height <- num_row_groups * 4 + + # Set the limits for the color scale, ignoring NA (negative values are now NA) + min_count <- min(heatmap_data_long$fitness, na.rm = TRUE) + max_count <- max(heatmap_data_long$fitness, na.rm = TRUE) + max_position <- max(heatmap_data$position) + + # Create the heatmap plot with explicit handling for positions > max_position + heatmap_plot <- ggplot(heatmap_data_long, aes(x = wt_aa_pos, y = mut_aa, fill = fitness)) + + scale_fill_gradientn( + colours = c("#D73027", "#F0F0F0", "#4575B4"), # From low = red to high = blue + values = c(0, abs(min_count) / (abs(min_count) + max_count), 1), # Manually define proportions + na.value = "grey35", # Color for NA values + limits = c(min_count, max_count) # Define the range for the color scale + ) + + scale_x_discrete(labels = function(x) { + numeric_pos <- as.numeric(gsub("[^0-9]", "", x)) + ifelse(numeric_pos > max_position, " ", x) + }) + + geom_tile() + + + # Add diagonal lines for synonymous mutations using geom_segment + geom_segment(data = syn_positions[syn_positions$position <= max_position, ], + aes(x = x - 0.485, xend = x + 0.485, + y = y - 0.485, yend = y + 0.485, color = synonymous), + size = 0.2) + + + # Manuelle Farbskala für die diagonalen Linien + scale_color_manual(values = c("TRUE" = "grey10"), labels = c("TRUE" = "")) + + + theme_minimal() + + labs(title = "Heatmap of Fitness Estimations for Mutations", x = "Wild-type Amino Acid", y = "Mutant Amino Acid", fill = "Fitness", color = "Synonymous Mutation") + + theme(plot.title = element_text(size = 16, face = "bold"), + axis.text.x = element_text(angle = 90, hjust = 1, vjust = 0.5, size = 12), + axis.text.y = element_text(size = 12), # Larger y-axis labels + axis.title.x = element_text(size = 16), + axis.title.y = element_text(size = 16), + legend.title = element_text(size = 14), # Larger legend title + legend.text = element_text(size = 12), # Larger legend text + panel.spacing = unit(0.1, "lines"), # Adjust panel spacing + strip.text = element_blank(), # Remove row group labels (facet numbers) + strip.background = element_blank(), + panel.grid.major = element_blank(), # Remove major grid lines + panel.grid.minor = element_blank()) + # Remove minor grid lines + facet_wrap(~ row_group, scales = "free_x", ncol = 1) + # Group by 75 positions per row + theme(panel.spacing = unit(0.2, "lines")) + + heatmap_plot <- heatmap_plot + + geom_point(data = heatmap_data_long, aes(size = ""), colour = "black", alpha = 0) # Invisible points for legend + heatmap_plot <- heatmap_plot + + guides(size = guide_legend(paste("Dropout (Counts <", threshold, ")"), override.aes = list(shape = 15, size = 8, colour = "grey35", alpha = 1))) # Define Legend for Dropouts + + # Save the heatmap plot + if (img_format == "pdf") { + ggsave(output_pdf_path, plot = heatmap_plot, width = 16, height = plot_height, dpi = 150, device = cairo_pdf) + } else { + ggsave(output_pdf_path, plot = heatmap_plot, width = 16, height = plot_height, dpi = 150) + } + + if (file.exists(output_pdf_path)) { + print("Heatmap image successfully created!") + } else { + print("Error: Heatmap image was not created.") + } +} + +# Test the function +fitness_heatmap( + "/Users/benjaminwehnert/CRG/DMS_QC/howard/prepared_gatk_for_fitness.csv", + "/Users/benjaminwehnert/CRG/DMS_QC/howard/fitness_heatmap.pdf", + threshold = 3 +) + +#test on gid1a data +fitness_heatmap( + "/Users/benjaminwehnert/Downloads/dimsum_fitness_gid1a/prepared_gatk_for_fitness.csv", + "/Users/benjaminwehnert/Downloads/dimsum_fitness_gid1a/fitness_heatmap.pdf", + threshold = 3 +) diff --git a/modules/local/dmsanalysis/bin/gatk_to_dimsum.R b/modules/local/dmsanalysis/bin/gatk_to_dimsum.R new file mode 100644 index 0000000..916404d --- /dev/null +++ b/modules/local/dmsanalysis/bin/gatk_to_dimsum.R @@ -0,0 +1,180 @@ +# input: wt_seq_path, input-gatk-counts filtered by codon library, output-gatk-counts filtered by codon library, start-stop-codon +# output: merged counts file for input and output data that can be fed into DiMSum +# sums up all synonymous mutations, assigning them to the wildtype sequence in the first line + + +# suppressMessages(library(seqinr)) +# suppressMessages(library(Biostrings)) + +# generate_dimsum_input <- function(wt_seq_path, gatk_input, gatk_output, pos_range) { +# # Parse the position range +# positions <- unlist(strsplit(pos_range, "-")) +# start_pos <- as.numeric(positions[1]) +# stop_pos <- as.numeric(positions[2]) +# +# # Load the wild-type sequence +# seq_data <- Biostrings::readDNAStringSet(filepath = wt_seq_path) +# wt_seq <- seq_data[[1]] # Extract the sequence +# wt_seq <- subseq(wt_seq, start = start_pos, end = stop_pos) +# +# # Convert wt_seq to a character string +# wt_seq <- as.character(wt_seq) +# +# # Split the wild-type sequence into codons (groups of 3 bases) +# wt_codons <- substring(wt_seq, seq(1, nchar(wt_seq), 3), seq(3, nchar(wt_seq), 3)) +# +# # Helper function to process GATK CSVs into count data +# process_gatk_file <- function(gatk_csv) { +# # Load the input GATK CSV file +# gatk_data <- read.csv(gatk_csv, stringsAsFactors = FALSE) +# +# # Calculate the sum of all synonymous mutation counts +# synonymous_counts <- sum(gatk_data$counts[grep("^S:", gatk_data$aa_mut)]) +# +# # Initialize a data frame with the wild-type sequence and its synonymous counts +# results <- data.frame( +# nt_seq = wt_seq, +# count = synonymous_counts, +# stringsAsFactors = FALSE +# ) +# +# # Iterate over each row in the input data +# for (i in 1:nrow(gatk_data)) { +# # Extract the mutation info +# codon_mut <- gatk_data$codon_mut[i] +# counts <- gatk_data$counts[i] +# +# # Create a mutable copy of the wild-type codons +# mutated_codons <- wt_codons +# +# # Apply the mutation +# mutations <- strsplit(codon_mut, ", ")[[1]] +# for (mutation in mutations) { +# codon_position <- as.numeric(sub(":.*", "", mutation)) +# new_codon <- sub(".*>", "", mutation) +# # Replace the codon at the specified position +# mutated_codons[codon_position] <- new_codon +# } +# +# # Convert the mutated codons back to a sequence string +# mutated_seq_string <- paste(mutated_codons, collapse = "") +# +# # Add the result to the data frame +# results <- rbind(results, data.frame(nt_seq = mutated_seq_string, count = counts)) +# } +# +# return(results) +# } +# +# # Process the GATK input and output files +# cat("Processing GATK input file...\n") +# input_data <- process_gatk_file(gatk_input) +# colnames(input_data)[2] <- "input1" # Rename count column +# +# cat("Processing GATK output file...\n") +# output_data <- process_gatk_file(gatk_output) +# colnames(output_data)[2] <- "output1" # Rename count column +# +# # Merge the input and output data +# merged_data <- merge(input_data, output_data, by = "nt_seq", all = TRUE) +# merged_data[is.na(merged_data)] <- 0 # Replace NA with 0 for missing counts +# +# # Write the merged data to a file +# output_file <- "merged_counts_for_dimsum.txt" +# write.table(merged_data, file = output_file, sep = "\t", row.names = FALSE, col.names = TRUE, quote = FALSE) +# cat("Merged counts file created:", output_file, "\n") +# } +# + +# +# generate_dimsum_input( +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/MORtn5_reference.fa", +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library.csv", +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library_dummy.csv", +# "23-1225" +# ) + + +suppressMessages(library(Biostrings)) + +generate_dimsum_input <- function(wt_seq_path, gatk_file, pos_range, output_file_path) { + # Parse the position range + positions <- unlist(strsplit(pos_range, "-")) + start_pos <- as.numeric(positions[1]) + stop_pos <- as.numeric(positions[2]) + + # Load the wild-type sequence + seq_data <- Biostrings::readDNAStringSet(filepath = wt_seq_path) + wt_seq <- seq_data[[1]] # Extract the sequence + wt_seq <- subseq(wt_seq, start = start_pos, end = stop_pos) + + # Convert wt_seq to a character string + wt_seq <- as.character(wt_seq) + + # Split the wild-type sequence into codons (groups of 3 bases) + wt_codons <- substring(wt_seq, seq(1, nchar(wt_seq), 3), seq(3, nchar(wt_seq), 3)) + + # Helper function to process GATK CSVs into count data + process_gatk_file <- function(gatk_csv) { + # Load the input GATK CSV file + gatk_data <- read.csv(gatk_csv, stringsAsFactors = FALSE) + + # Initialize a data frame for results + results <- data.frame( + nt_seq = character(), + count = numeric(), + stringsAsFactors = FALSE + ) + + # Iterate over each row in the input data + for (i in 1:nrow(gatk_data)) { + # Extract the mutation info + codon_mut <- gatk_data$codon_mut[i] + counts <- gatk_data$counts[i] + + # Create a mutable copy of the wild-type codons + mutated_codons <- wt_codons + + # Apply the mutation + mutations <- strsplit(codon_mut, ", ")[[1]] + for (mutation in mutations) { + codon_position <- as.numeric(sub(":.*", "", mutation)) + new_codon <- sub(".*>", "", mutation) + # Replace the codon at the specified position + mutated_codons[codon_position] <- new_codon + } + + # Convert the mutated codons back to a sequence string + mutated_seq_string <- paste(mutated_codons, collapse = "") + + # Add the result to the data frame + results <- rbind(results, data.frame(nt_seq = mutated_seq_string, count = counts)) + } + + return(results) + } + + # Process the GATK file + cat("Processing GATK file...\n") + processed_data <- process_gatk_file(gatk_file) + + # Write the processed data to a file without column names + write.table(processed_data, file = output_file_path, sep = "\t", row.names = FALSE, col.names = FALSE, quote = FALSE) +} + +# generate_dimsum_input( +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/MORtn5_reference.fa", +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library.csv", +# "23-1225", +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/dimsum_input.tsv" +# ) + + + + + + + + + + diff --git a/modules/local/dmsanalysis/bin/global_position_biases_counts_and_counts_per_cov.R b/modules/local/dmsanalysis/bin/global_position_biases_counts_and_counts_per_cov.R new file mode 100644 index 0000000..142e1c9 --- /dev/null +++ b/modules/local/dmsanalysis/bin/global_position_biases_counts_and_counts_per_cov.R @@ -0,0 +1,163 @@ +# input: prefiltered gatk path, aa seq path, window_size (sliding window), output_path_folder +# output: two lineplots showing counts & counts per coverage divided in types of variants (single-/double-/triple-base exchange) + +library(zoo) # sliding window +library(dplyr) +library(ggplot2) +library(scales) + +position_biases <- function(prefiltered_gatk_path, aa_seq_path, window_size = 10) { + + # Load and process the data + prefiltered_gatk <- read.table(prefiltered_gatk_path, sep = ",", fill = NA, header = TRUE) + prefiltered_gatk$pos <- as.numeric(sub("(\\D)(\\d+)(\\D)", "\\2", prefiltered_gatk$pos_mut)) + unique_pos <- unique(as.numeric(prefiltered_gatk$pos)) + aa_seq <- readLines(aa_seq_path, warn = FALSE) + aa_seq_length <- nchar(aa_seq) + aa_positions <- seq(nchar(aa_seq)) + + means_counts_single <- rep(NA, nchar(aa_seq)) + means_counts_double <- rep(NA, nchar(aa_seq)) + means_counts_triple <- rep(NA, nchar(aa_seq)) + means_counts_per_cov_single <- rep(NA, nchar(aa_seq)) + means_counts_per_cov_double <- rep(NA, nchar(aa_seq)) + means_counts_per_cov_triple <- rep(NA, nchar(aa_seq)) + + + # Loop through each position in the amino acid sequence + for (i in 1:(nchar(aa_seq))) { + + # Filter the data for the current position in aa_positions + window_data <- prefiltered_gatk %>% filter(prefiltered_gatk$pos %in% aa_positions[i]) + + # Calculate mean for Single mutations (where varying_bases == 1) + window_data_single <- window_data %>% filter(varying_bases == 1) + means_counts_single[i] <- mean(window_data_single$counts, na.rm = FALSE) + means_counts_per_cov_single[i] <- mean(window_data_single$counts_per_cov, na.rm = FALSE) + + # Calculate mean for Double mutations (where varying_bases == 2) + window_data_double <- window_data %>% filter(varying_bases == 2) + means_counts_double[i] <- mean(window_data_double$counts, na.rm = FALSE) + means_counts_per_cov_double[i] <- mean(window_data_double$counts_per_cov, na.rm = FALSE) + + # Calculate mean for Triple mutations (where varying_bases == 3) + window_data_triple <- window_data %>% filter(varying_bases == 3) + means_counts_triple[i] <- mean(window_data_triple$counts, na.rm = FALSE) + means_counts_per_cov_triple[i] <- mean(window_data_triple$counts_per_cov, na.rm = FALSE) + } + + pos_bias_df <- data.frame(pos = seq(nchar(aa_seq))) + + + pos_bias_df$rolling_mean_counts_single <- rollapply(means_counts_single, width = window_size, + FUN = function(x) (mean(x, na.rm = TRUE) + 0.00001), fill = "extend") + pos_bias_df$rolling_mean_counts_double <- rollapply(means_counts_double, width = window_size, + FUN = function(x) (mean(x, na.rm = TRUE) + 0.00001), fill = "extend") + pos_bias_df$rolling_mean_counts_triple <- rollapply(means_counts_triple, width = window_size, + FUN = function(x) (mean(x, na.rm = TRUE) + 0.00001), fill = "extend") + + pos_bias_df$rolling_mean_counts_per_cov_single <- rollapply(means_counts_per_cov_single, width = window_size, + FUN = function(x) (mean(x, na.rm = TRUE) + 0.00001), fill = "extend") + pos_bias_df$rolling_mean_counts_per_cov_double <- rollapply(means_counts_per_cov_double, width = window_size, + FUN = function(x) (mean(x, na.rm = TRUE) + 0.00001), fill = "extend") + pos_bias_df$rolling_mean_counts_per_cov_triple <- rollapply(means_counts_per_cov_triple, width = window_size, + FUN = function(x) (mean(x, na.rm = TRUE) + 0.00001), fill = "extend") + + + # Replace NAs with 0 for rolling means and SE + pos_bias_df$rolling_mean_counts_single[is.na(pos_bias_df$rolling_mean_counts_single)] <- 0.00001 + pos_bias_df$rolling_mean_counts_double[is.na(pos_bias_df$rolling_mean_counts_double)] <- 0.00001 + pos_bias_df$rolling_mean_counts_triple[is.na(pos_bias_df$rolling_mean_counts_triple)] <- 0.00001 + pos_bias_df$rolling_mean_counts_per_cov_single[is.na(pos_bias_df$rolling_mean_counts_per_cov_single)] <- 0.000001 + pos_bias_df$rolling_mean_counts_per_cov_double[is.na(pos_bias_df$rolling_mean_counts_per_cov_double)] <- 0.000001 + pos_bias_df$rolling_mean_counts_per_cov_triple[is.na(pos_bias_df$rolling_mean_counts_per_cov_triple)] <- 0.000001 + + + + plots_theme <- list( + + # Add the minimal theme to the list + theme_minimal(), + + # Customize legend title and appearance + theme(legend.title = element_text(size = 10, face = "bold"), + legend.position = "right"), + + # Customize guides for the legend elements + guides( + color = guide_legend(title = "Mutation Type", order = 1), # Title for line color + fill = guide_legend(title = "Standard Deviation", order = 2), # Title for ribbon fill + linetype = guide_legend(title = "Required Coverage", order = 3) # Title for line type + ) + ) + + + # Placeholder name for the linetype + linetype_placeholder <- "Required coverage" + + rolling_counts_plot <- ggplot(pos_bias_df, aes(x = pos)) + + + # Add line for rolling mean coverage + geom_line(aes(y = rolling_mean_counts_single, color = "One Varying Base")) + + geom_line(aes(y = rolling_mean_counts_double, color = "Two Varying Bases")) + + geom_line(aes(y = rolling_mean_counts_triple, color = "Three Varying Bases")) + + + # Individual axis labels for this plot + xlab("Amino Acid Position") + + ylab("Counts") + + + # Manually set color and fill labels + scale_color_manual(name = "Mutation Type", + values = c("One Varying Base" = "chocolate", "Two Varying Bases" = "darkolivegreen3", "Three Varying Bases" = "deepskyblue1"), + limits = c("One Varying Base", "Two Varying Bases", "Three Varying Bases")) + # Color for line + + # Apply the saved theme and design elements at the end + plots_theme + + + scale_y_continuous(trans = 'log10', labels = scales::comma) + + + + + rolling_counts_per_cov_plot <- ggplot(pos_bias_df, aes(x = pos)) + + + # Add line for rolling mean coverage + geom_line(aes(y = rolling_mean_counts_per_cov_single, color = "One Varying Base")) + + geom_line(aes(y = rolling_mean_counts_per_cov_double, color = "Two Varying Bases")) + + geom_line(aes(y = rolling_mean_counts_per_cov_triple, color = "Three Varying Bases")) + + + # Individual axis labels for this plot + xlab("Amino Acid Position") + + ylab("Counts") + + + # Manually set color and fill labels + scale_color_manual(name = "Mutation Type", + values = c("One Varying Base" = "chocolate", "Two Varying Bases" = "darkolivegreen3", "Three Varying Bases" = "deepskyblue1"), + limits = c("One Varying Base", "Two Varying Bases", "Three Varying Bases")) + # Color and legend order for lines + + # Apply the saved theme and design elements at the end + plots_theme + + + scale_y_continuous(trans = 'log10', labels = scales::comma) + + + ggsave(filename = "rolling_counts.pdf", plot = rolling_counts_plot, device = "pdf", width = 10, height = 6) + ggsave(filename = "rolling_counts_per_cov.pdf", plot = rolling_counts_per_cov_plot, device = "pdf", width = 10, height = 6) +} + +# Example call to the function +#position_biases("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library.csv", "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/aa_seq.txt", window_size = 18, output_path_folder = "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs") + + + + + + + + + + + + + + diff --git a/modules/local/dmsanalysis/bin/global_position_biases_cov.R b/modules/local/dmsanalysis/bin/global_position_biases_cov.R new file mode 100644 index 0000000..857f4f2 --- /dev/null +++ b/modules/local/dmsanalysis/bin/global_position_biases_cov.R @@ -0,0 +1,86 @@ +# input: prefiltered gatk path, aa seq path, window_size (sliding window), output_path_folder, aimed counts per aa variant +# output: lineplot showing coverage over positions and dotted line: Assuming all potential 21 variants are equally distributed, this is the coverage one need to get the aimed counts per variant. + +library(zoo) # sliding window +library(dplyr) +library(ggplot2) +library(scales) +position_biases <- function(prefiltered_gatk_path, aa_seq_path, window_size = 10, output_file_path, targeted_counts_per_aa_variant = 100) { + + # Load and process the data + prefiltered_gatk <- read.table(prefiltered_gatk_path, sep = ",", fill = NA, header = TRUE) + prefiltered_gatk$pos <- as.numeric(sub("(\\D)(\\d+)(\\D)", "\\2", prefiltered_gatk$pos_mut)) + unique_pos <- unique(as.numeric(prefiltered_gatk$pos)) + aa_seq <- readLines(aa_seq_path, warn = FALSE) + aa_seq_length <- nchar(aa_seq) + aa_positions <- seq(nchar(aa_seq)) + + means_cov <- rep(NA, nchar(aa_seq)) + + # Calculate means for cov (should be the same over all variants in one position) + for (i in 1:(nchar(aa_seq))) { + window_data <- prefiltered_gatk %>% filter(prefiltered_gatk$pos %in% aa_positions[i]) + means_cov[i] <- mean(window_data$cov, na.rm = FALSE) + } + + pos_bias_df <- data.frame(pos = seq(nchar(aa_seq))) + + # Log-transform the rolling means to avoid issues with zeros (log(y + 0.001)) + pos_bias_df$rolling_mean_cov <- rollapply(means_cov, width = window_size, + FUN = function(x) (mean(x, na.rm = TRUE)), fill = "extend") + + # Generate the plot + plots_theme <- list( + + # Customize legend appearance (leave titles blank) + guides( + color = guide_legend(title = NULL, order = 1), # Title for line color + fill = guide_legend(title = NULL, order = 2), # Title for ribbon fill + linetype = guide_legend(title = NULL, order = 3) # Title for line type + ), + + # Apply minimal theme + theme_minimal() + ) + + + linetype_placeholder <- "Required coverage" + + rolling_cov_plot <- ggplot(pos_bias_df, aes(x = pos, y = rolling_mean_cov)) + + + # Add line for rolling mean coverage + geom_line(aes(color = "Coverage")) + + + # Add a horizontal line with a mapped linetype so it appears in the legend + geom_hline(aes(yintercept = ((21 * nchar(aa_seq) * targeted_counts_per_aa_variant)), + linetype = linetype_placeholder), + color = "black", linewidth = 0.3) + + + # Individual axis labels for this plot + xlab("Amino Acid Position") + + ylab("Coverage") + + + # Manually set color and fill labels + scale_color_manual(values = c("Coverage" = "black")) + # Color for line + + # Set the linetype with a label that uses the targeted threshold dynamically + scale_linetype_manual(values = c("Required coverage" = "dotted"), + labels = paste("Required coverage \n for", as.character(targeted_counts_per_aa_variant), "counts per variant \n if equally present")) + + + # Apply the saved theme and design elements at the end + plots_theme + + + # Set y-axis to log scale and apply comma formatting + scale_y_continuous(trans = 'log10', labels = scales::comma) + + + # Return the plot + ggsave(filename = output_file_path, plot = rolling_cov_plot, device = "pdf", width = 10, height = 6) +} + +# Example call to the function +#position_biases("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library.csv", "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/aa_seq.txt", window_size = 18, output_path_folder = "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/rolling_coverage.pdf", targeted_counts_per_aa_variant = 15) + + + + diff --git a/modules/local/dmsanalysis/bin/install_packages.R b/modules/local/dmsanalysis/bin/install_packages.R new file mode 100644 index 0000000..2f98481 --- /dev/null +++ b/modules/local/dmsanalysis/bin/install_packages.R @@ -0,0 +1,82 @@ +# List of required packages +required_packages <- c("Biostrings", "dplyr", "ggplot2", "scales", "zoo", "reshape2", "tidyr", "tidyverse") + +# Function to install and load missing packages +install_if_missing <- function(pkg) { + # Check if package is already installed + if (!requireNamespace(pkg, quietly = TRUE)) { + message(paste("Installing package:", pkg)) + + # Ensure BiocManager is installed for Bioconductor packages + if (!requireNamespace("BiocManager", quietly = TRUE)) { + install.packages("BiocManager", repos = "https://cloud.r-project.org") # Set a CRAN mirror + } + + # Install Bioconductor packages via BiocManager + if (pkg %in% c("Biostrings", "GenomicRanges", "IRanges")) { # Add more Bioconductor packages if needed + BiocManager::install(pkg) + } else { + # Install CRAN packages + install.packages(pkg, repos = "https://cloud.r-project.org") + } + } else { + message(paste("Package", pkg, "is already installed")) + } +} + +# Install and load all required packages +invisible(lapply(required_packages, install_if_missing)) + +message("All required packages are installed and loaded.") + + + + + + + + + + + + + + + + + + + +##Find current versions of dependencies: +# List of your packages +packages <- c("dplyr", "ggplot2", "scales", "zoo", "reshape2", "tidyr", "tidyverse", "Biostrings") + +# Function to get the current version of each package +get_package_version <- function(pkg) { + if (requireNamespace(pkg, quietly = TRUE)) { + return(packageVersion(pkg)) + } else { + return(NA) + } +} + +# Check and print versions +package_versions <- sapply(packages, get_package_version) +print(package_versions) + + + +# Check the version of libcurl +system("curl --version") + +# Check the version of OpenSSL +system("openssl version") + +# Check the version of libxml2 +system("xml2-config --version") + +# Check the version of zlib +system("zlib-flate -version") + +system("ldconfig -p | grep zlib") + diff --git a/modules/local/dmsanalysis/bin/logdiff.R b/modules/local/dmsanalysis/bin/logdiff.R new file mode 100644 index 0000000..5982d6d --- /dev/null +++ b/modules/local/dmsanalysis/bin/logdiff.R @@ -0,0 +1,149 @@ + +# Maybe develop additional ideas to characterise variants below 10 percentile -> Do we find patterns that help the user to understand why certain variants are hard to count? + +# input: completed_prefiltered_gatk path, output folder path +# output: two logdiff-plots (counts_per_cov): 1st lineplot 2nd dotplot showing type of variant (varying bases - 1/2/3) + +library(dplyr) +library(ggplot2) +library(scales) + + logdiff_plot <- function(prefiltered_gatk_path) { + + # Load the data + prefiltered_gatk <- read.table(prefiltered_gatk_path, sep = ",", fill = NA, header = TRUE) + + # Sort by counts_per_cov while keeping corresponding varying_bases + sorted_counts <- prefiltered_gatk %>% + arrange(counts_per_cov) # Sort by counts_per_cov + + # Create a new column for the sorted index (1, 2, 3, ...) + sorted_counts$ids <- 1:nrow(sorted_counts) + + # Calculate Q1 (10% quantile) and Q3 (90% quantile) + Q1 <- quantile(sorted_counts$counts_per_cov, 0.10, na.rm = TRUE) + Q3 <- quantile(sorted_counts$counts_per_cov, 0.90, na.rm = TRUE) + + # Calculate the LogDiff + LogDiff <- log10(Q3) - log10(Q1) + + # Create the first plot with a line + line_plot <- ggplot(sorted_counts, aes(x = ids, y = counts_per_cov)) + + geom_line(color = "black") + + + # Set axis labels + xlab("Variants") + + ylab("Counts per Coverage") + + + # Apply logarithmic scale to the y-axis + scale_y_continuous(trans = 'log10') + + + # Add horizontal dotted lines at Q1 and Q3 + geom_hline(yintercept = Q1, linetype = "dotted", color = "black") + + geom_hline(yintercept = Q3, linetype = "dotted", color = "black") + + + # Add the LogDiff value to the top left corner + annotate("text", x = 0, y = max(sorted_counts$counts_per_cov), label = paste("LogDiff =", round(LogDiff, 2)), hjust = 0, vjust = 1, size = 5, color = "black") + + + # Apply the minimal theme + theme_minimal() + + # Save the line plot as a PDF + ggsave(filename = "logdiff_plot.pdf", plot = line_plot, device = "pdf", width = 10, height = 6) + + # Create the second plot with colored dots based on varying_bases + colored_plot <- ggplot(sorted_counts, aes(x = ids, y = counts_per_cov, color = as.factor(varying_bases))) + + + # Add horizontal dotted lines at Q1 and Q3 + geom_hline(yintercept = Q1, linetype = "dotted", color = "black") + + geom_hline(yintercept = Q3, linetype = "dotted", color = "black") + + + geom_point(size = 0.9) + # Use points instead of lines + + # Set axis labels + xlab("Variants") + + ylab("Counts per Coverage") + + + # Apply logarithmic scale to the y-axis + scale_y_continuous(trans = 'log10') + + + # Add the LogDiff value to the top left corner + annotate("text", x = 1, y = max(sorted_counts$counts_per_cov), label = paste("LogDiff =", round(LogDiff, 2)), hjust = 0, vjust = 1, size = 5, color = "black") + + + # Add color scale for varying_bases + scale_color_manual(values = c("1" = "chocolate", "2" = "darkolivegreen3", "3" = "deepskyblue1"), name = "Varying \n Bases") + + + # Apply the minimal theme + theme_minimal() + + # Save the colored plot as a PDF + ggsave(filename = "logdiff_varying_bases.pdf", plot = colored_plot, device = "pdf", width = 10, height = 6) + } + + # Example call of the function +# logdiff_plot("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/completed_prefiltered_gatk.csv", "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs") + + + + + + # logdiff_plot_codon_mut <- function(prefiltered_gatk_path, output_folder_path) { + # + # # Load the data + # prefiltered_gatk <- read.table(prefiltered_gatk_path, sep = ",", fill = NA, header = TRUE) + # + # # Extract the codon number from the codon_mut column (e.g., '377' from '377:TCC>CTG') + # prefiltered_gatk <- prefiltered_gatk %>% + # mutate(position = as.numeric(gsub(":.*", "", codon_mut))) # Extract the number before the colon + # + # # Sort by counts_per_cov while keeping corresponding position + # sorted_counts <- prefiltered_gatk %>% + # arrange(counts_per_cov) # Sort by counts_per_cov + # + # # Create a new column for the sorted index (1, 2, 3, ...) + # sorted_counts$ids <- 1:nrow(sorted_counts) + # + # # Calculate Q1 (10% quantile) and Q3 (90% quantile) + # Q1 <- quantile(sorted_counts$counts_per_cov, 0.10, na.rm = TRUE) + # Q3 <- quantile(sorted_counts$counts_per_cov, 0.90, na.rm = TRUE) + # + # # Calculate the LogDiff + # LogDiff <- log10(Q3) - log10(Q1) + # + # # Create the second plot with a larger gray area and preserved horizontal lines and LogDiff + # codon_mut_plot <- ggplot(sorted_counts, aes(x = ids, y = counts_per_cov, color = position)) + + # + # # Add horizontal dotted lines in the background + # geom_hline(yintercept = Q1, linetype = "dotted", color = "black") + + # geom_hline(yintercept = Q3, linetype = "dotted", color = "black") + + # + # # Add points with smaller size + # geom_point(size = 0.9) + + # + # # Set axis labels + # xlab("Sorted Index") + + # ylab("Counts per Coverage") + + # + # # Apply logarithmic scale to the y-axis + # scale_y_continuous(trans = 'log10', labels = scales::comma) + + # + # # Add the LogDiff value to the top left corner + # annotate("text", x = 1, y = max(sorted_counts$counts_per_cov), label = paste("LogDiff =", round(LogDiff, 2)), hjust = 0, vjust = 1, size = 5, color = "black") + + # + # # Add a color gradient with a larger gray area + # scale_color_gradient2(low = "blue", mid = "lightgray", high = "green", midpoint = median(sorted_counts$position, na.rm = TRUE), name = "Position") + + # + # # Apply the minimal theme + # theme_minimal() + # + # # Save the second plot as a PDF + # ggsave(filename = paste(output_folder_path, "/logdiff_position.pdf", sep = ""), + # plot = codon_mut_plot, device = "pdf", width = 10, height = 6) + # } + # + # # Example call + # logdiff_plot_codon_mut("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/completed_prefiltered_gatk.csv", "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs") + # + + + diff --git a/modules/local/dmsanalysis/bin/low_count_variants.R b/modules/local/dmsanalysis/bin/low_count_variants.R new file mode 100644 index 0000000..ef1902b --- /dev/null +++ b/modules/local/dmsanalysis/bin/low_count_variants.R @@ -0,0 +1,130 @@ +#!!!UNFINISHED!!! --> Decide if those graphics make sense for users. If so, further develop them and put everything in a function :) + + +library(tidyverse) + +completed_prefiltered_gatk <- read.csv("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/completed_prefiltered_gatk.csv") +View(completed_prefiltered_gatk) + +# Lade die notwendigen Pakete +library(dplyr) +library(tidyr) + +# Funktion zur Klassifizierung der Basenwechsel +classify_base_changes <- function(wt_codon, variant_codon) { + # Definition der Purine und Pyrimidine + purines <- c("A", "G") + pyrimidines <- c("C", "T") + + # Initialisiere die Zähler + pur_pur <- 0 + pur_pyr <- 0 + pyr_pur <- 0 + pyr_pyr <- 0 + + # Vergleiche jede Base zwischen wt_codon und variant_codon + for (i in 1:3) { + wt_base <- substr(wt_codon, i, i) + var_base <- substr(variant_codon, i, i) + + # Klassifiziere den Basenwechsel + if (wt_base %in% purines && var_base %in% purines) { + pur_pur <- pur_pur + 1 + } else if (wt_base %in% purines && var_base %in% pyrimidines) { + pur_pyr <- pur_pyr + 1 + } else if (wt_base %in% pyrimidines && var_base %in% purines) { + pyr_pur <- pyr_pur + 1 + } else if (wt_base %in% pyrimidines && var_base %in% pyrimidines) { + pyr_pyr <- pyr_pyr + 1 + } + } + + # Erstelle eine Ausgabe als DataFrame + return(data.frame(pur_pur, pur_pyr, pyr_pur, pyr_pyr)) +} + +# Beispielanwendung auf den Datensatz +completed_prefiltered_gatk <- completed_prefiltered_gatk %>% + rowwise() %>% + mutate(base_change_classification = list(classify_base_changes(wt_codon, Variant))) %>% + unnest(base_change_classification) + + + + + + + + +# Load necessary packages +library(dplyr) +library(ggplot2) + +# 1. Data preparation +# Create a binary variable for low vs. high counts_per_cov +threshold <- quantile(completed_prefiltered_gatk$counts_per_cov, 0.10, na.rm = TRUE) +completed_prefiltered_gatk <- completed_prefiltered_gatk %>% + mutate(group = ifelse(counts_per_cov < threshold, "Bottom 10%", "Top 90%")) + +# 2. Calculate the mean frequency for each category in both groups +frequency_data <- completed_prefiltered_gatk %>% + group_by(group) %>% + summarise( + pur_pur_mean = mean(pur_pur, na.rm = TRUE), + pur_pyr_mean = mean(pur_pyr, na.rm = TRUE), + pyr_pur_mean = mean(pyr_pur, na.rm = TRUE), + pyr_pyr_mean = mean(pyr_pyr, na.rm = TRUE) + ) %>% + pivot_longer(cols = c(pur_pur_mean, pur_pyr_mean, pyr_pur_mean, pyr_pyr_mean), + names_to = "mutation_type", + values_to = "mean_frequency") %>% + # Ensure that all mutation types are present + complete(mutation_type, group, fill = list(mean_frequency = 0)) + +# 3. Visualize the distribution of frequencies for the top 90% and bottom 10% +ggplot(frequency_data, aes(x = mutation_type, y = mean_frequency, fill = group)) + + geom_bar(stat = "identity", position = "dodge") + + labs(x = "Mutation Type", y = "Mean Frequency", title = "Distribution of Mutation Types for Top 90% vs. Bottom 10%") + + scale_fill_manual(values = c("Top 90%" = "steelblue", "Bottom 10%" = "tomato")) + + theme_minimal() + + scale_x_discrete(labels = c("pur_pur" = "Purine > Purine", "pur_pyr" = "Purine > Pyrimidine", + "pyr_pur" = "Pyrimidine > Purine", "pyr_pyr" = "Pyrimidine > Pyrimidine")) + + + + + + + + + + +threshold <- quantile(completed_prefiltered_gatk$counts_per_cov, 0.10, na.rm = TRUE) +completed_prefiltered_gatk <- completed_prefiltered_gatk %>% + mutate(group = ifelse(counts_per_cov < threshold, "Bottom 10%", "Top 90%")) + +# 2. Create a flag for variants with at least one pur_pyr or pur_pur +completed_prefiltered_gatk <- completed_prefiltered_gatk %>% + mutate(has_pur_mutation = ifelse(pyr_pur > 0 | pur_pyr > 0, 1, 0)) + +# 3. Calculate the proportion of variants with at least one pur_pyr or pur_pur for each group +proportion_data <- completed_prefiltered_gatk %>% + group_by(group) %>% + summarise( + total_variants = n(), + variants_with_pur_mutation = sum(has_pur_mutation), + proportion_with_pur_mutation = variants_with_pur_mutation / total_variants + ) + +# 4. Print the results +print(proportion_data) + +# 5. Optional: Visualize the proportion for each group +ggplot(proportion_data, aes(x = group, y = proportion_with_pur_mutation, fill = group)) + + geom_bar(stat = "identity") + + labs(x = "Group", y = "Proportion of Variants", title = "Proportion of Variants with at Least One Pur_pyr or Pur_pur Mutation") + + scale_fill_manual(values = c("Top 90%" = "steelblue", "Bottom 10%" = "tomato")) + + theme_minimal() + + + diff --git a/modules/local/dmsanalysis/bin/possible_mutations.R b/modules/local/dmsanalysis/bin/possible_mutations.R new file mode 100644 index 0000000..1206d58 --- /dev/null +++ b/modules/local/dmsanalysis/bin/possible_mutations.R @@ -0,0 +1,105 @@ +# input: wildtype-seq (string or fasta file), start&stopp pos., output_folder_path, mutagenesis_type (choose from nnk, nns, max_diff_to_wt, custom), if you choose custom, add: custom_codon_library -> comma-separated .txt (-> "AAA, AAC, AAG, AAT, ...") +# output: .csv with all possible programmed codons for each position + +suppressMessages(library(Biostrings)) + +# Define the function +generate_possible_variants <- function(wt_seq_input, start_stop_pos, mutagenesis_type = "nnk", + custom_codon_library = NULL, output_file) { + # Parse the start and stop positions from the input format "start-stop" + positions <- unlist(strsplit(start_stop_pos, "-")) + start_pos <- as.numeric(positions[1]) + stop_pos <- as.numeric(positions[2]) + + # Check if the input is a file or a string + if (file.exists(wt_seq_input)) { + # If it's a file, read the sequence from the fasta file + seq_data <- readDNAStringSet(filepath = wt_seq_input) + wt_seq <- seq_data[[1]] # Extract the sequence + } else { + # Otherwise, treat the input as a sequence string + wt_seq <- DNAString(wt_seq_input) + } + + # Extract the sequence between start and stop codons + coding_seq <- subseq(wt_seq, start = start_pos, end = stop_pos) + coding_seq <- as.character(coding_seq) + + # List of predefined NNK & NNS codons + nnk_codons <- c('AAG', 'AAT', 'ATG', 'ATT', 'AGG', 'AGT', 'ACG', 'ACT', + 'TAG', 'TAT', 'TTG', 'TTT', 'TGG', 'TGT', 'TCG', 'TCT', + 'GAG', 'GAT', 'GTG', 'GTT', 'GGG', 'GGT', 'GCG', 'GCT', + 'CAG', 'CAT', 'CTG', 'CTT', 'CGG', 'CGT', 'CCG', 'CCT') + + nns_codons <- c('AAG', 'AAC', 'ATG', 'ATC', 'AGG', 'AGC', 'ACG', 'ACC', + 'TAG', 'TAC', 'TTG', 'TTC', 'TGG', 'TGC', 'TCG', 'TCC', + 'GAG', 'GAC', 'GTG', 'GTC', 'GGG', 'GGC', 'GCG', 'GCC', + 'CAG', 'CAC', 'CTG', 'CTC', 'CGG', 'CGC', 'CCG', 'CCC') + + # Function to split a DNA sequence into codons (triplets) + split_into_codons <- function(seq) { + return(strsplit(seq, "(?<=.{3})", perl = TRUE)[[1]]) + } + + # Read custom codons if mode is 'custom' + if (mutagenesis_type == "custom") { + if (is.null(custom_codon_library) || !file.exists(custom_codon_library)) { + stop("Custom codons file must be provided and valid when using 'custom' mutagenesis_type") + } + # Read and parse the custom codons from the file + custom_codons <- unlist(strsplit(readLines(custom_codon_library), ",")) + custom_codons <- trimws(custom_codons) # Remove any whitespace + } + + # Split wild-type sequence into codons + wt_codons <- split_into_codons(coding_seq) + + # Initialize DataFrame to store mutated variants + result <- data.frame(Codon_Number = integer(), wt_codon = character(), Variant = character(), stringsAsFactors = FALSE) + + # Determine the codon list based on the mutagenesis_type + get_codon_list <- function(wt_codon) { + if (mutagenesis_type == "nnk") { + return(nnk_codons) + } else if (mutagenesis_type == "nns") { + return(nns_codons) + } else if (mutagenesis_type == "max_diff_to_wt") { + if (substr(wt_codon, 3, 3) == "T") { + return(nns_codons) + } else { + return(nnk_codons) + } + } else if (mutagenesis_type == "custom") { + return(custom_codons) + } else { + stop("Invalid mutagenesis_type Choose from 'nnk', 'nns', 'max_diff_to_wt', or 'custom'.") + } + } + + # Iterate over each codon in the wild-type sequence + for (i in seq_along(wt_codons)) { + wt_codon <- wt_codons[i] + codon_list <- get_codon_list(wt_codon) + + # Filter codons that are different from the wild-type codon + possible_variants <- codon_list[codon_list != wt_codon] + + # Add all variants to the result list, including the wild-type codon + for (variant in possible_variants) { + result <- rbind(result, data.frame(Codon_Number = i, wt_codon = wt_codon, Variant = variant, stringsAsFactors = FALSE)) + } + } + + # Save the variants into a CSV file + write.csv(result, output_file, row.names = FALSE) +} + +# Example usage +# Possibly generate a custom codons file: "AAA, AAC, AAG, AAT, ..." +# generate_possible_variants("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/MORtn5_reference.fa", "23-1225", "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/possible_NNK_mutations.csv", mutagenesis_type = "nnk") ### this one's correct for the dataset +# generate_possible_variants("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/MORtn5_reference.fa", "23-1225", "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/possible_NNK_mutations_taylors_nnk_and_nns.csv", mutagenesis_type = "max_diff_to_wt") + + + + + diff --git a/modules/local/dmsanalysis/bin/prepare_gatk_data_for_count_heatmaps.R b/modules/local/dmsanalysis/bin/prepare_gatk_data_for_count_heatmaps.R new file mode 100644 index 0000000..694a1fa --- /dev/null +++ b/modules/local/dmsanalysis/bin/prepare_gatk_data_for_count_heatmaps.R @@ -0,0 +1,82 @@ +# input: prefiltered GATK path (filtered for codon library), aa-seq file path, output path, threshold (for minimum counts to recognize variant) +# output: csv file serving as basis for counts_per_cov_heatmap function + +suppressMessages(library(dplyr)) +suppressMessages(library(ggplot2)) +suppressMessages(library(tidyr)) +suppressMessages(library(reshape2)) +suppressMessages(library(scales)) + +prepare_gatk_data_for_counts_heatmaps <- function(gatk_file_path, aa_seq_file_path, output_csv_path, threshold = 3) { + # Load the raw GATK data + raw_gatk <- read.table(gatk_file_path, sep = ",", header = TRUE) + + # Read the wild-type amino acid sequence from the text file + wt_seq <- readLines(aa_seq_file_path) + wt_seq <- unlist(strsplit(wt_seq, "")) # Split the sequence into individual amino acids + + # Summarize counts-per-cov for each unique aa mutation in pos_mut + aggregated_data <- raw_gatk %>% + group_by(pos_mut) %>% + summarize(total_counts_per_cov = sum(counts_per_cov, na.rm = TRUE), + total_counts = sum(counts, na.rm = TRUE)) # Also sum the counts + + # Extract the wild-type position and mutations from 'pos_mut' + aggregated_data <- aggregated_data %>% + mutate( + wt_aa = sub("(\\D)(\\d+)(\\D)", "\\1", pos_mut), # Wild-type amino acid (e.g., S) + position = as.numeric(sub("(\\D)(\\d+)(\\D)", "\\2", pos_mut)), # Position (e.g., 3) + mut_aa = sub("(\\D)(\\d+)(\\D)", "\\3", pos_mut) # Mutant amino acid (e.g., R) + ) + + # Replace 'X' with '*', indicating the stop codon + aggregated_data <- aggregated_data %>% + mutate(mut_aa = ifelse(mut_aa == "X", "*", mut_aa)) + + # Replace 'X' with '*' in the wild-type amino acid sequence as well + wt_seq <- ifelse(wt_seq == "X", "*", wt_seq) + + # Define all 20 standard amino acids and the stop codon "*" + all_amino_acids <- c("A", "C", "D", "E", "F", "G", "H", "I", "K", "L", + "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y", "*") + + # Create a list of all positions in the wild-type sequence + all_positions <- 1:length(wt_seq) + + # Create a complete grid of all possible combinations of positions and amino acids + complete_data <- expand.grid(mut_aa = all_amino_acids, position = all_positions) + + # Merge the summarized data with the complete grid (filling missing entries with 0) + heatmap_data <- complete_data %>% + left_join(aggregated_data, by = c("mut_aa", "position")) %>% + mutate(total_counts_per_cov = ifelse(is.na(total_counts_per_cov), 0, total_counts_per_cov), + wt_aa = wt_seq[position]) # Assign the wild-type amino acid + + # Set variants with counts < threshold to NA + heatmap_data <- heatmap_data %>% + mutate( + total_counts_per_cov = ifelse(total_counts < threshold, NA, total_counts_per_cov), + total_counts = ifelse(total_counts < threshold, NA, total_counts) + ) + + # Fill pos_mut column + heatmap_data <- heatmap_data %>% + mutate( + pos_mut = ifelse(is.na(pos_mut), + paste0(wt_aa, position, mut_aa), + pos_mut) + ) + + # Save the aggregated data to a CSV file + write.csv(heatmap_data, file = output_csv_path, row.names = FALSE) + print(paste("Aggregated data saved to:", output_csv_path)) +} + + +# Aufruf der Datenaufbereitungsfunktion +# prepare_gatk_data_for_counts_heatmaps( +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library.csv", +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/aa_seq.txt", +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/prepared_gatk_data.csv", +# threshold = 3 +# ) diff --git a/modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R b/modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R new file mode 100644 index 0000000..ef25ce9 --- /dev/null +++ b/modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R @@ -0,0 +1,134 @@ +suppressMessages(library(dplyr)) + +prepare_gatk_data_for_fitness_heatmap <- function(csv_file_path, aa_seq_path, dimsum_fitness_path, output_csv_path) { + # Load the CSV file + gatk_data <- read.csv(csv_file_path) + + # Read the wild-type amino acid sequence from the text file + aa_seq <- readLines(aa_seq_path) + aa_seq <- unlist(strsplit(aa_seq, "")) # Split the sequence into individual amino acids + + # Add the mutated sequence column + gatk_data <- gatk_data %>% + mutate( + mutated_sequence = sapply(1:nrow(gatk_data), function(i) { + # Create a copy of the wild-type sequence + mutated_seq <- aa_seq + # Extract mutation information + position <- as.numeric(position[i]) + mut_aa <- mut_aa[i] + # Apply the mutation + if (!is.na(mut_aa) && position > 0 && position <= length(mutated_seq)) { + mutated_seq[position] <- mut_aa + } + # Return the mutated sequence as a string + paste(mutated_seq, collapse = "") + }) + ) + + # # Save the updated data to a new CSV file + # write.csv(gatk_data, file = output_csv_path, row.names = FALSE) + # print(paste("Updated data with mutated sequences saved to:", output_csv_path)) + + + load(dimsum_fitness_path) + dimsum_fitness <- rbind(all_variants, synonymous) + dimsum_fitness <- dimsum_fitness[-which(dimsum_fitness$WT == T)[-1],] # remove duplicate + rm(doubles,singles,all_variants,synonymous,wildtype) + + # Perform a left join to retain all rows from gatk_data + merged_data <- gatk_data %>% + left_join(dimsum_fitness, by = c("mutated_sequence" = "aa_seq")) + + # Handle wild-type rows specifically + merged_data <- merged_data %>% + mutate( + # If WT from dimsum_fitness exists, use it; otherwise keep as is + WT = ifelse(is.na(WT) & mutated_sequence == paste(aa_seq, collapse = ""), TRUE, WT) + ) + + # Save the merged data to a CSV file + write.csv(merged_data, file = output_csv_path, row.names = FALSE) + print(paste("Merged data saved to:", output_csv_path)) +} + + + + +# add_mutated_sequence("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/prepared_gatk_data.csv", +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/aa_seq.txt", +# +# "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/testing_outputs/prepared_gatk_for_fitness.csv") + + + + + + +suppressMessages(library(dplyr)) + +add_mutated_sequence <- function(csv_file_path, aa_seq_path, dimsum_fitness_path, output_csv_path) { + + # Load the CSV file + gatk_data <- read.csv(csv_file_path) + + # Read the wild-type amino acid sequence from the text file + aa_seq <- readLines(aa_seq_path) + aa_seq <- unlist(strsplit(aa_seq, "")) # Split the sequence into individual amino acids + + # Add the mutated sequence column + gatk_data <- gatk_data %>% + mutate( + mutated_sequence = sapply(1:nrow(gatk_data), function(i) { + # Create a copy of the wild-type sequence + mutated_seq <- aa_seq + # Extract mutation information + position <- as.numeric(gatk_data$position[i]) + mut_aa <- gatk_data$mut_aa[i] + # Apply the mutation + if (!is.na(mut_aa) && position > 0 && position <= length(mutated_seq)) { + mutated_seq[position] <- mut_aa + } + # Return the mutated sequence as a string + paste(mutated_seq, collapse = "") + }) + ) + + # Load dimsum_fitness data + load(dimsum_fitness_path) + dimsum_fitness <- rbind(all_variants, synonymous) + + # Remove unnecessary columns + dimsum_fitness <- dimsum_fitness %>% + select(-nt_seq, -Nham_nt, -Nmut_codons, -indel, -STOP, -STOP_readthrough) + + # Ensure one-to-one mapping for the merge + dimsum_fitness <- dimsum_fitness %>% + distinct(aa_seq, .keep_all = TRUE) # Keep only one row per unique `aa_seq` + + # Perform a left join to merge gatk_data with dimsum_fitness + merged_data <- gatk_data %>% + left_join(dimsum_fitness, by = c("mutated_sequence" = "aa_seq")) + + # Save the merged data to a CSV file + write.csv(merged_data, file = output_csv_path, row.names = FALSE) + print(paste("Merged data saved to:", output_csv_path)) +} + + + + + + + +# test data +prepare_gatk_data_for_fitness_heatmap("/Users/benjaminwehnert/CRG/DMS_QC/howard/bin4/intermediate_files/processed_gatk_files/variantCounts_for_heatmaps.csv", + "/Users/benjaminwehnert/CRG/DMS_QC/howard/bin4/intermediate_files/aa_seq.txt", + "/Users/benjaminwehnert/CRG/DMS_QC/howard/dimsum/howards_data/howards_data_fitness_replicates.RData", + "/Users/benjaminwehnert/CRG/DMS_QC/howard/prepared_gatk_for_fitness.csv") + +#test on gid1a data +prepare_gatk_data_for_fitness_heatmap("/Users/benjaminwehnert/Downloads/variantCounts_for_heatmaps.csv", + "/Users/benjaminwehnert/Downloads/aa_seq.txt", + "/Users/benjaminwehnert/Downloads/2025-01-20_GID1A_DiMSum_1_fitness_replicates.RData", + "/Users/benjaminwehnert/Downloads/dimsum_fitness_gid1a/prepared_gatk_for_fitness.csv") diff --git a/modules/local/dmsanalysis/bin/process_raw_gatk.R b/modules/local/dmsanalysis/bin/process_raw_gatk.R new file mode 100644 index 0000000..292fa6c --- /dev/null +++ b/modules/local/dmsanalysis/bin/process_raw_gatk.R @@ -0,0 +1,46 @@ +# input: gatk variantcounts tsv file, output_path +# output: csv with column names. Creates additional counts_per_cov column. Fills pos_mut column for synonymous mutations. Sorted out variants that have mutations, but do not show up in the specifying columns -> was affecting roughly 30 low-count variants out of over 15000 in Taylor's data + +library("dplyr") +process_raw_gatk <- function(gatk_file_path, output_csv_path) { + + # Set the column names + colnames <- c("counts", "cov", "mean_length_variant_reads", "varying_bases", + "base_mut", "varying_codons", "codon_mut", "aa_mut", "pos_mut") + + # Read the GATK file into a data frame + gatk_raw <- read.table(gatk_file_path, sep = "\t", header = FALSE, fill = TRUE, col.names = colnames) + + # Filter out rows where 'aa_mut' is empty or NA + gatk_raw <- gatk_raw[!(gatk_raw$aa_mut == "" | is.na(gatk_raw$aa_mut)), ] + + # Handle synonymous mutations: where aa_mut starts with "S" and pos_mut is either NA or "" + gatk_raw <- gatk_raw %>% + rowwise() %>% + mutate( + pos_mut = ifelse( + (is.na(pos_mut) | pos_mut == "") & grepl("^S:", aa_mut), + # Construct the new 'pos_mut' entry for synonymous mutations + paste0( + sub("S:([A-Z])>[A-Z]", "\\1", aa_mut), # Get the original amino acid from 'aa_mut' + sub("^(\\d+):.*", "\\1", codon_mut), # Get the position from 'codon_mut' + sub("S:[A-Z]>([A-Z])", "\\1", aa_mut) # Get the mutated amino acid from 'aa_mut' + ), + pos_mut # Keep the existing 'pos_mut' if it's not NA or "" + ) + ) %>% + ungroup() %>% + mutate(counts_per_cov = counts / cov) + + # Write the cleaned data frame to a CSV file + write.csv(gatk_raw, file = output_csv_path, row.names = FALSE) +} + + + + +# Example usage (can be used for testing): +# process_raw_gatk("/path/to/gatk_file.txt", "/path/to/output_file.csv") +#process_raw_gatk("/Users/benjaminwehnert/CRG/DMS_QC/testing_data/output_premerged_vsearch.variantCounts", "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/raw_gatk.csv") + + From de75c7a861ead5d1e1270726fe8341edc23463af Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 15:12:54 +0200 Subject: [PATCH 080/317] Update dmscore.nf --- workflows/dmscore.nf | 5 +++-- 1 file changed, 3 insertions(+), 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 55e3fd6..37e611e 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -10,6 +10,7 @@ include { BWA_MEM } from '../modules/nf-core/bwa/mem/main' include { BAMFILTER_DMS } from '../modules/local/bamfilteringdms' include { PREMERGE } from '../modules/local/premerge' include { GATK_SATURATIONMUTAGENESIS } from '../modules/local/gatk/saturationmutagenesis' +include { DMSANALYSIS_AASEQ } from '../modules/local/dmsanalysis/aaseq' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -130,12 +131,12 @@ workflow DMSCORE { BWA_MEM.out.bam ) - PREMERGE( + PREMERGE ( BAMFILTER_DMS.out.bam, ch_fasta.map{ it[1] } // extract fasta path from Tuple ) - GATK_SATURATIONMUTAGENESIS( + GATK_SATURATIONMUTAGENESIS ( PREMERGE.out.bam, // merged reads (tuple(meta, merged_reads.fastq)) ch_fasta.map{ it[1] }, // fasta path (path) reading_frame_ch, // codon range (path) From 39e84a70572d4af9f9962aed4d68df6586e024ce Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 15:14:49 +0200 Subject: [PATCH 081/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index 43e639c..6d2a2ca 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -2,12 +2,12 @@ process DMSANALYSIS_AASEQ { tag "$meta.id" label 'process_single' - // Define the correct container and/or conda environment + conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:8fa94107068d5af9" input: tuple val(meta), path(wt_seq) - path pos_range + val pos_range output: tuple val(meta), path("aa_seq.txt"), emit: aa_seq @@ -19,7 +19,7 @@ process DMSANALYSIS_AASEQ { script: def prefix = task.ext.prefix ?: "${meta.id}" """ - start_stop_codon=\$(cat $pos_range) + start_stop_codon="$pos_range" Rscript -e "source('../bin/aa_seq.R'); aa_seq('$wt_seq', '\$start_stop_codon', 'aa_seq.txt')" From 683ddcf291fd3293aac7ba414c01a692b40724e6 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 15:20:12 +0200 Subject: [PATCH 082/317] Update dmscore.nf --- workflows/dmscore.nf | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 37e611e..f592fb4 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -143,6 +143,11 @@ workflow DMSCORE { min_counts_ch // min_counts (val) ) + DMSANALYSIS_AASEQ ( + ch_fasta, + reading_frame_ch + ) + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From b3d27982fc44a088ca27799aae6573f984cef192 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 15:24:49 +0200 Subject: [PATCH 083/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 1 - 1 file changed, 1 deletion(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index 6d2a2ca..4ace6e2 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -17,7 +17,6 @@ process DMSANALYSIS_AASEQ { task.ext.when == null || task.ext.when script: - def prefix = task.ext.prefix ?: "${meta.id}" """ start_stop_codon="$pos_range" From bec122d1b4487ec2d4dce30365b554631c0cdfdd Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 15:27:19 +0200 Subject: [PATCH 084/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 1 - 1 file changed, 1 deletion(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index 4ace6e2..a3df06e 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -1,5 +1,4 @@ process DMSANALYSIS_AASEQ { - tag "$meta.id" label 'process_single' conda "${moduleDir}/environment.yml" From 3bb52951b5db0570decf39439712bb79f125e579 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 15:49:20 +0200 Subject: [PATCH 085/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index a3df06e..027b0bb 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -7,6 +7,7 @@ process DMSANALYSIS_AASEQ { input: tuple val(meta), path(wt_seq) val pos_range + path script // aa_seq.R output: tuple val(meta), path("aa_seq.txt"), emit: aa_seq @@ -19,7 +20,7 @@ process DMSANALYSIS_AASEQ { """ start_stop_codon="$pos_range" - Rscript -e "source('../bin/aa_seq.R'); aa_seq('$wt_seq', '\$start_stop_codon', 'aa_seq.txt')" + Rscript -e "source('$script'); aa_seq('$wt_seq', '\$start_stop_codon', 'aa_seq.txt')" cat <<-END_VERSIONS > versions.yml "\${task.process}": From 659794a8dba2d08d9ef0e726773cdab36559ffa4 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 15:52:07 +0200 Subject: [PATCH 086/317] Update dmscore.nf --- workflows/dmscore.nf | 9 ++++++++- 1 file changed, 8 insertions(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index f592fb4..972ed54 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -39,6 +39,12 @@ Channel .value(params.min_counts) .set { min_counts_ch } +// Define R scripts as channels +Channel + .fromPath("modules/local/dmsanalysis/bin/aa_seq.R", checkIfExists: true) + .set { aa_seq_script_ch } + + workflow DMSCORE { @@ -145,7 +151,8 @@ workflow DMSCORE { DMSANALYSIS_AASEQ ( ch_fasta, - reading_frame_ch + reading_frame_ch, + aa_seq_script_ch // path to aa_seq.R (defined at the top) ) emit: From ba955d29bec73f883f008630712f175a80be6f41 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 15:54:47 +0200 Subject: [PATCH 087/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index 027b0bb..f037519 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -2,7 +2,7 @@ process DMSANALYSIS_AASEQ { label 'process_single' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:8fa94107068d5af9" + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" input: tuple val(meta), path(wt_seq) From 38c6d3a893ac221f8d28957ae565ee353419706f Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 16:05:47 +0200 Subject: [PATCH 088/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index f037519..12096a5 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -23,8 +23,8 @@ process DMSANALYSIS_AASEQ { Rscript -e "source('$script'); aa_seq('$wt_seq', '\$start_stop_codon', 'aa_seq.txt')" cat <<-END_VERSIONS > versions.yml - "\${task.process}": - r: \$(R --version | head -n 1 | sed 's/^R version //') + "DMSANALYSIS_AASEQ": + r: $(R --version | head -n 1 | sed 's/^R version //') aa_seq_script: custom_R_script_bin END_VERSIONS """ From 7347161d84ce4f1d041b3814a6c710bc79ae6e93 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 16:09:21 +0200 Subject: [PATCH 089/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index 12096a5..9ca8d6b 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -27,6 +27,7 @@ process DMSANALYSIS_AASEQ { r: $(R --version | head -n 1 | sed 's/^R version //') aa_seq_script: custom_R_script_bin END_VERSIONS + """ stub: From 8248af99bade6f77c5a1f3ca00d101c136ef29da Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 16:12:10 +0200 Subject: [PATCH 090/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index 9ca8d6b..c936b08 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -24,10 +24,9 @@ process DMSANALYSIS_AASEQ { cat <<-END_VERSIONS > versions.yml "DMSANALYSIS_AASEQ": - r: $(R --version | head -n 1 | sed 's/^R version //') + r: \$\(R --version | head -n 1 | sed 's/^R version //'\) aa_seq_script: custom_R_script_bin END_VERSIONS - """ stub: From c6b5f8ae98546b8911326141c44970dc06eadb94 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 16:15:22 +0200 Subject: [PATCH 091/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 8 +++++--- 1 file changed, 5 insertions(+), 3 deletions(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index c936b08..35d2c02 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -19,12 +19,14 @@ process DMSANALYSIS_AASEQ { script: """ start_stop_codon="$pos_range" - + + R_version=\$(R --version | head -n 1 | sed 's/^R version //') + Rscript -e "source('$script'); aa_seq('$wt_seq', '\$start_stop_codon', 'aa_seq.txt')" - + cat <<-END_VERSIONS > versions.yml "DMSANALYSIS_AASEQ": - r: \$\(R --version | head -n 1 | sed 's/^R version //'\) + r: \$R_version aa_seq_script: custom_R_script_bin END_VERSIONS """ From 5c3c1a71b3e3f0afc7dee75d3881cae643b56e87 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 2 May 2025 16:36:15 +0200 Subject: [PATCH 092/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index 35d2c02..fc17294 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -1,4 +1,5 @@ process DMSANALYSIS_AASEQ { + tag "amino_acid_sequence" label 'process_single' conda "${moduleDir}/environment.yml" From a2d4c463813f9a0df59bf20d36d9edc3393df5d6 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 6 May 2025 13:37:27 +0200 Subject: [PATCH 093/317] Update dmscore.nf --- workflows/dmscore.nf | 22 +++++++++++++++++++--- 1 file changed, 19 insertions(+), 3 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 972ed54..bc8379b 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -40,9 +40,25 @@ Channel .set { min_counts_ch } // Define R scripts as channels -Channel - .fromPath("modules/local/dmsanalysis/bin/aa_seq.R", checkIfExists: true) - .set { aa_seq_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/SeqDepth_simulation.R", checkIfExists: true).set { seqdepth_simulation_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/aa_seq.R", checkIfExists: true).set { aa_seq_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/complete_prefiltered_gatk.R", checkIfExists: true).set { complete_gatk_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/counts_heatmap.R", checkIfExists: true).set { counts_heatmap_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R", checkIfExists: true).set { counts_per_cov_heatmap_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/detect_codons.R", checkIfExists: true).set { detect_codons_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/filter_gatk_by_codon_library.R", checkIfExists: true).set { filter_by_library_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/fitness_heatmap.R", checkIfExists: true).set { fitness_heatmap_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/gatk_to_dimsum.R", checkIfExists: true).set { gatk_to_dimsum_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/global_position_biases_counts_and_counts_per_cov.R", checkIfExists: true).set { global_bias_counts_cov_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/global_position_biases_cov.R", checkIfExists: true).set { global_bias_cov_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/install_packages.R", checkIfExists: true).set { install_packages_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/logdiff.R", checkIfExists: true).set { logdiff_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/low_count_variants.R", checkIfExists: true).set { low_count_variants_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/possible_mutations.R", checkIfExists: true).set { possible_mutations_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/prepare_gatk_data_for_count_heatmaps.R", checkIfExists: true).set { prepare_counts_heatmap_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R", checkIfExists: true).set { prepare_fitness_heatmap_script_ch } +Channel.fromPath("modules/local/dmsanalysis/bin/process_raw_gatk.R", checkIfExists: true).set { process_raw_gatk_script_ch } + From 9239de8d50e1d3185637ac3a3679b3b31c4d66cf Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Tue, 6 May 2025 15:50:41 +0200 Subject: [PATCH 094/317] possible mutations module --- log.txt | 428 +++++++++++++++++++++++++++++ modules/local/possiblemutations.nf | 89 ++++++ 2 files changed, 517 insertions(+) create mode 100644 log.txt create mode 100644 modules/local/possiblemutations.nf diff --git a/log.txt b/log.txt new file mode 100644 index 0000000..c55d577 --- /dev/null +++ b/log.txt @@ -0,0 +1,428 @@ +May-02 15:57:46.796 [main] DEBUG nextflow.cli.Launcher - $> nextflow run . -profile test,docker --fasta /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta --reading_frame 352-1383 --min_counts 2 --outdir ./results -resume +May-02 15:57:46.924 [main] DEBUG nextflow.cli.CmdRun - N E X T F L O W ~ version 24.04.4 +May-02 15:57:46.948 [main] DEBUG nextflow.plugin.PluginsFacade - Setting up plugin manager > mode=prod; embedded=false; plugins-dir=/Users/benjaminwehnert/.nextflow/plugins; core-plugins: nf-amazon@2.5.3,nf-azure@1.6.1,nf-cloudcache@0.4.1,nf-codecommit@0.2.1,nf-console@1.1.3,nf-ga4gh@1.3.0,nf-google@1.13.2-patch1,nf-tower@1.9.1,nf-wave@1.4.2-patch1 +May-02 15:57:46.960 [main] INFO o.pf4j.DefaultPluginStatusProvider - Enabled plugins: [] +May-02 15:57:46.961 [main] INFO o.pf4j.DefaultPluginStatusProvider - Disabled plugins: [] +May-02 15:57:46.963 [main] INFO org.pf4j.DefaultPluginManager - PF4J version 3.12.0 in 'deployment' mode +May-02 15:57:46.979 [main] INFO org.pf4j.AbstractPluginManager - No plugins +May-02 15:57:47.853 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` +May-02 15:57:47.871 [main] DEBUG nextflow.config.ConfigBuilder - Found config local: /Users/benjaminwehnert/dmscore/nextflow.config +May-02 15:57:47.879 [main] DEBUG nextflow.config.ConfigBuilder - Parsing config file: /Users/benjaminwehnert/dmscore/nextflow.config +May-02 15:57:47.897 [main] DEBUG n.secret.LocalSecretsProvider - Secrets store: /Users/benjaminwehnert/.nextflow/secrets/store.json +May-02 15:57:47.900 [main] DEBUG nextflow.secret.SecretsLoader - Discovered secrets providers: [nextflow.secret.LocalSecretsProvider@7ee8130e] - activable => nextflow.secret.LocalSecretsProvider@7ee8130e +May-02 15:57:47.910 [main] DEBUG nextflow.config.ConfigBuilder - Applying config profile: `test,docker` +May-02 15:57:49.363 [main] DEBUG nextflow.config.ConfigBuilder - Available config profiles: [bih, cfc_dev, uzl_omics, ifb_core, embl_hd, denbi_qbic, alice, mjolnir_globe, uppmax, giga, incliva, ilifu, ki_luria, uge, icr_alma, rosalind_uge, lugh, mccleary, unibe_ibu, vai, czbiohub_aws, jax, roslin, ccga_med, tes, scw, unc_longleaf, tigem, tubingen_apg, google, apollo, ipop_up, vsc_calcua, pdc_kth, googlels, ceci_nic5, humantechnopole, stjude, daisybio, eddie, medair, biowulf, apptainer, bi, bigpurple, adcra, cedars, pawsey_setonix, vsc_kul_uhasselt, pawsey_nimbus, ucl_myriad, utd_ganymede, charliecloud, seattlechildrens, icr_davros, ceres, arm, munin, rosalind, hasta, cfc, uzh, shu_bmrc, ebi_codon_slurm, ebc, ccga_dx, crick, ku_sund_danhead, marvin, shifter, biohpc_gen, mana, mamba, york_viking, unc_lccc, wehi, awsbatch, wustl_htcf, arcc, ceci_dragon2, imperial, maestro, software_license, cannon, genotoul, nci_gadi, abims, janelia, nu_genomics, googlebatch, oist, sahmri, kaust, alliance_canada, mpcdf, leicester, vsc_ugent, create, sage, cambridge, jex, podman, ebi_codon, cheaha, xanadu, nyu_hpc, test, marjorie, computerome, ucd_sonic, seg_globe, sanger, dkfz, bluebear, pasteur, einstein, ethz_euler, m3c, test_full, imb, ucl_cscluster, tuos_stanage, azurebatch, hki, seadragon, crukmi, csiro_petrichor, qmul_apocrita, wave, docker, engaging, gis, hypatia, psmn, eva, unity, cropdiversityhpc, nygc, fgcz, conda, crg, singularity, mpcdf_viper, pe2, self_hosted_runner, tufts, uw_hyak_pedslabs, binac2, debug, genouest, cbe, unsw_katana, gitpod, phoenix, seawulf, uod_hpc, fub_curta, uct_hpc, aws_tower, binac, fsu_draco] +May-02 15:57:49.415 [main] DEBUG nextflow.cli.CmdRun - Applied DSL=2 by global default +May-02 15:57:49.433 [main] DEBUG nextflow.cli.CmdRun - Launching `./main.nf` [golden_curran] DSL2 - revision: 84101fc51c +May-02 15:57:49.435 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins declared=[nf-schema@2.3.0] +May-02 15:57:49.435 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins default=[] +May-02 15:57:49.436 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins resolved requirement=[nf-schema@2.3.0] +May-02 15:57:49.436 [main] DEBUG nextflow.plugin.PluginUpdater - Installing plugin nf-schema version: 2.3.0 +May-02 15:57:49.444 [main] INFO org.pf4j.AbstractPluginManager - Plugin 'nf-schema@2.3.0' resolved +May-02 15:57:49.444 [main] INFO org.pf4j.AbstractPluginManager - Start plugin 'nf-schema@2.3.0' +May-02 15:57:49.450 [main] DEBUG nextflow.plugin.BasePlugin - Plugin started nf-schema@2.3.0 +May-02 15:57:49.513 [main] DEBUG nextflow.Session - Session UUID: 36458275-f468-409a-bb18-98996facae84 +May-02 15:57:49.513 [main] DEBUG nextflow.Session - Run name: golden_curran +May-02 15:57:49.513 [main] DEBUG nextflow.Session - Executor pool size: 8 +May-02 15:57:49.521 [main] DEBUG nextflow.file.FilePorter - File porter settings maxRetries=3; maxTransfers=50; pollTimeout=null +May-02 15:57:49.531 [main] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'FileTransfer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +May-02 15:57:49.553 [main] DEBUG nextflow.cli.CmdRun - + Version: 24.04.4 build 5917 + Created: 01-08-2024 07:05 UTC (09:05 CEST) + System: Mac OS X 15.0 + Runtime: Groovy 4.0.21 on OpenJDK 64-Bit Server VM 17.0.13+0 + Encoding: UTF-8 (UTF-8) + Process: 36172@MacBook-Air-von-Benjamin.local [127.0.0.1] + CPUs: 8 - Mem: 8 GB (162.4 MB) - Swap: 8 GB (449.1 MB) +May-02 15:57:49.575 [main] DEBUG nextflow.Session - Work-dir: /Users/benjaminwehnert/dmscore/work [Mac OS X] +May-02 15:57:49.575 [main] DEBUG nextflow.Session - Script base path does not exist or is not a directory: /Users/benjaminwehnert/dmscore/bin +May-02 15:57:49.591 [main] DEBUG nextflow.executor.ExecutorFactory - Extension executors providers=[] +May-02 15:57:49.599 [main] DEBUG nextflow.Session - Observer factory: DefaultObserverFactory +May-02 15:57:49.619 [main] DEBUG nextflow.Session - Observer factory: ValidationObserverFactory +May-02 15:57:49.650 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` +May-02 15:57:49.675 [main] DEBUG nextflow.cache.CacheFactory - Using Nextflow cache factory: nextflow.cache.DefaultCacheFactory +May-02 15:57:49.685 [main] DEBUG nextflow.util.CustomThreadPool - Creating default thread pool > poolSize: 9; maxThreads: 1000 +May-02 15:57:49.783 [main] DEBUG nextflow.Session - Session start +May-02 15:57:49.785 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow started -- trace file: /Users/benjaminwehnert/dmscore/results/pipeline_info/execution_trace_2025-05-02_15-57-48.txt +May-02 15:57:49.944 [main] DEBUG nextflow.script.ScriptRunner - > Launching execution +May-02 15:57:51.084 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +May-02 15:57:51.555 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema +May-02 15:57:51.556 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema +May-02 15:57:51.559 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +May-02 15:57:51.560 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema +May-02 15:57:52.021 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema +May-02 15:57:52.022 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema +May-02 15:57:52.024 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +May-02 15:57:52.025 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema +May-02 15:57:52.227 [main] INFO nextflow.Nextflow - +------------------------------------------------------ + ,--./,-. + ___ __ __ __ ___ /,-._.--~' + |\ | |__ __ / ` / \ |__) |__ } { + | \| | \__, \__/ | \ |___ \`-._,-`-, + `._,._,' + nf-core/dmscore 1.0.0dev +------------------------------------------------------ +Input/output options + input : https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv + outdir : ./results + min_counts : 2 + +Reference genome options + genome : R64-1-1 + fasta : /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta + +Institutional config options + config_profile_name : Test profile + config_profile_description: Minimal test dataset to check pipeline function + +Generic options + trace_report_suffix : 2025-05-02_15-57-48 + +Core Nextflow options + runName : golden_curran + containerEngine : docker + launchDir : /Users/benjaminwehnert/dmscore + workDir : /Users/benjaminwehnert/dmscore/work + projectDir : /Users/benjaminwehnert/dmscore + userName : benjaminwehnert + profile : test,docker + configFiles : /Users/benjaminwehnert/dmscore/nextflow.config + +!! Only displaying parameters that differ from the pipeline defaults !! +------------------------------------------------------ +* The nf-core framework + https://doi.org/10.1038/s41587-020-0439-x + +* Software dependencies + https://github.com/nf-core/dmscore/blob/master/CITATIONS.md + +May-02 15:57:52.229 [main] DEBUG n.validation.ValidationExtension - Starting parameters validation +May-02 15:57:52.604 [main] DEBUG nextflow.validation.SchemaEvaluator - Started validating /BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv +May-02 15:57:53.970 [main] DEBUG nextflow.validation.SchemaEvaluator - Validation of file 'https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv' passed! +May-02 15:57:54.378 [main] DEBUG n.v.FormatDirectoryPathEvaluator - Cloud blob storage paths are not supported by 'FormatDirectoryPathEvaluator': 's3://ngi-igenomes/igenomes/' +May-02 15:57:54.380 [main] DEBUG n.validation.ValidationExtension - Finishing parameters validation +May-02 15:57:54.462 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:FASTQC +May-02 15:57:54.464 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:FASTQC` matches process NFCORE_DMSCORE:DMSCORE:FASTQC +May-02 15:57:54.477 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-02 15:57:54.477 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-02 15:57:54.483 [main] DEBUG nextflow.executor.Executor - [warm up] executor > local +May-02 15:57:54.488 [main] DEBUG n.processor.LocalPollingMonitor - Creating local task monitor for executor 'local' > cpus=8; memory=8 GB; capacity=8; pollInterval=100ms; dumpInterval=5m +May-02 15:57:54.490 [main] DEBUG n.processor.TaskPollingMonitor - >>> barrier register (monitor: local) +May-02 15:57:54.672 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:MULTIQC +May-02 15:57:54.673 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:MULTIQC` matches process NFCORE_DMSCORE:DMSCORE:MULTIQC +May-02 15:57:54.676 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-02 15:57:54.676 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-02 15:57:54.686 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BWA_INDEX +May-02 15:57:54.688 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-02 15:57:54.688 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-02 15:57:54.702 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_high` matches labels `process_high` for process with name NFCORE_DMSCORE:DMSCORE:BWA_MEM +May-02 15:57:54.706 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-02 15:57:54.706 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-02 15:57:54.715 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS +May-02 15:57:54.718 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-02 15:57:54.718 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-02 15:57:54.729 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:PREMERGE +May-02 15:57:54.732 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-02 15:57:54.732 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-02 15:57:54.746 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_high` matches labels `process_high` for process with name NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS +May-02 15:57:54.750 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-02 15:57:54.750 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-02 15:57:54.761 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ +May-02 15:57:54.764 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-02 15:57:54.764 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-02 15:57:54.781 [main] DEBUG nextflow.Session - Config process names validation disabled as requested +May-02 15:57:54.783 [main] DEBUG nextflow.Session - Igniting dataflow network (22) +May-02 15:57:54.790 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:FASTQC +May-02 15:57:54.792 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:MULTIQC +May-02 15:57:54.793 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX +May-02 15:57:54.794 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_MEM +May-02 15:57:54.794 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS +May-02 15:57:54.794 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:PREMERGE +May-02 15:57:54.795 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS +May-02 15:57:54.795 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ +May-02 15:57:54.797 [main] DEBUG nextflow.script.ScriptRunner - Parsed script files: + Script_e5f965e09aa3641b: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/premerge.nf + Script_113f045ba19c2c46: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/fastqc/main.nf + Script_4f612cfd8c52e8cd: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nextflow_pipeline/main.nf + Script_215a636da7ab24a2: /Users/benjaminwehnert/dmscore/./subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfschema_plugin/main.nf + Script_68606fccdecc54d9: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf + Script_f88958acfb50d604: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/dmsanalysis/aaseq.nf + Script_df773f5847ef0bc7: /Users/benjaminwehnert/dmscore/./workflows/dmscore.nf + Script_0f3077e98f6a93f6: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/mem/main.nf + Script_5c4e8d4051efa81e: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/nf-core/utils_nfcore_pipeline/main.nf + Script_4ff7366a79e0ef06: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/bamfilteringdms.nf + Script_c568aae5b239c1c5: /Users/benjaminwehnert/dmscore/main.nf + Script_4955288afd8ca61e: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/multiqc/main.nf + Script_37dcd664b2773148: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/gatk/saturationmutagenesis.nf + Script_25aa31c18d513c61: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/index/main.nf +May-02 15:57:54.797 [main] DEBUG nextflow.script.ScriptRunner - > Awaiting termination +May-02 15:57:54.798 [main] DEBUG nextflow.Session - Session await +May-02 15:57:54.893 [Actor Thread 10] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.002 s; external sort time: 0.016 s; total time: 0.018 s +May-02 15:57:54.893 [Actor Thread 1] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.002 s; external sort time: 0.016 s; total time: 0.018 s +May-02 15:57:54.894 [Actor Thread 1] DEBUG nextflow.file.FileCollector - >> temp file exists? false +May-02 15:57:54.894 [Actor Thread 10] DEBUG nextflow.file.FileCollector - >> temp file exists? false +May-02 15:57:54.895 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Missed collect-file cache -- cause: java.nio.file.NoSuchFileException: /Users/benjaminwehnert/dmscore/work/collect-file/9aa80ec1e5fc421feabac9f320c60e5e +May-02 15:57:54.895 [Actor Thread 10] DEBUG nextflow.file.FileCollector - Missed collect-file cache -- cause: java.nio.file.NoSuchFileException: /Users/benjaminwehnert/dmscore/work/collect-file/a9ac31ffb91d381d31c639634831f9b7 +May-02 15:57:54.901 [Actor Thread 10] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/a9ac31ffb91d381d31c639634831f9b7 +May-02 15:57:54.901 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/9aa80ec1e5fc421feabac9f320c60e5e +May-02 15:57:54.910 [Actor Thread 10] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-1468229241367644128 +May-02 15:57:54.910 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-9598669176738179442 +May-02 15:57:54.920 [Actor Thread 15] DEBUG nextflow.util.HashBuilder - [WARN] Unknown hashing type: class Script_df773f5847ef0bc7$_runScript_closure1$_closure7 +May-02 15:57:55.001 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-02 15:57:55.003 [Task submitter] INFO nextflow.Session - [a4/7e1d06] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta) +May-02 15:57:55.734 [FileTransfer-2] DEBUG nextflow.file.FilePorter - Local cache found for foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq at /Users/benjaminwehnert/dmscore/work/stage-36458275-f468-409a-bb18-98996facae84/cc/f938a8775ac8befd4337b2732c7dc9/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq +May-02 15:57:55.734 [FileTransfer-1] DEBUG nextflow.file.FilePorter - Local cache found for foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq at /Users/benjaminwehnert/dmscore/work/stage-36458275-f468-409a-bb18-98996facae84/28/abb932c7e69700944b1605c1ee72d2/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq +May-02 15:57:55.867 [Actor Thread 9] INFO nextflow.processor.TaskProcessor - [5a/035f68] Cached process > NFCORE_DMSCORE:DMSCORE:FASTQC (gid1a_1_quality_1_pe) +May-02 15:57:55.967 [Actor Thread 9] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'PublishDir' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +May-02 15:57:56.063 [Actor Thread 18] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 2; slices: 1; internal sort time: 0.022 s; external sort time: 0.002 s; total time: 0.024 s +May-02 15:57:56.063 [Actor Thread 18] DEBUG nextflow.file.FileCollector - >> temp file exists? true +May-02 15:57:56.065 [Actor Thread 18] DEBUG nextflow.file.FileCollector - Retrieved cached collect-files from: /Users/benjaminwehnert/dmscore/work/collect-file/0dde6dd9bfe7393330209153ab0f204f +May-02 15:57:56.070 [Actor Thread 18] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-5944054719096968617 +May-02 15:57:57.568 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 1; name: NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/a4/7e1d06298bb5191af0791289427bba] +May-02 15:57:57.569 [Task monitor] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'TaskFinalizer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +May-02 15:57:57.621 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-02 15:57:57.623 [Task submitter] INFO nextflow.Session - [09/593b38] Submitted process > NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1) +May-02 15:57:57.692 [Actor Thread 17] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process +May-02 15:59:23.448 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 2; name: NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1); status: COMPLETED; exit: 1; error: -; workDir: /Users/benjaminwehnert/dmscore/work/09/593b381f4e3da5089ff973e0ac9448] +May-02 15:59:23.464 [TaskFinalizer-2] DEBUG nextflow.processor.TaskProcessor - Handling unexpected condition for + task: name=NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1); work-dir=/Users/benjaminwehnert/dmscore/work/09/593b381f4e3da5089ff973e0ac9448 + error [nextflow.exception.ProcessFailedException]: Process `NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1)` terminated with an error exit status (1) +May-02 15:59:23.479 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-02 15:59:23.480 [Task submitter] INFO nextflow.Session - [0c/0721f6] Submitted process > NFCORE_DMSCORE:DMSCORE:MULTIQC +May-02 15:59:23.536 [TaskFinalizer-2] ERROR nextflow.processor.TaskProcessor - Error executing process > 'NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1)' + +Caused by: + Process `NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1)` terminated with an error exit status (1) + + +Command executed: + + start_stop_codon="352-1383" + + Rscript -e "source('aa_seq.R'); aa_seq('GID1A_SUNi_ref_small.fasta', '$start_stop_codon', 'aa_seq.txt')" + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + r: $(R --version | head -n 1 | sed 's/^R version //') + aa_seq_script: custom_R_script_bin + END_VERSIONS + +Command exit status: + 1 + +Command output: + (empty) + +Command error: + Unable to find image 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' locally + 0fd2e39a5bf2ecaa: Pulling from library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned + 6114410fdbce: Pulling fs layer + bc6461884445: Pulling fs layer + bc6461884445: Download complete + 6114410fdbce: Download complete + Digest: sha256:2e209ed7eb4e3c8c5f29a6e9f65dc2113c00e69b5c7dc4ff5bc5abbd2b080242 + Status: Downloaded newer image for community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa + .command.sh: line 12: "${task.process}": + r: $(R --version | head -n 1 | sed 's/^R version //') + aa_seq_script: custom_R_script_bin + : bad substitution + +Work dir: + /Users/benjaminwehnert/dmscore/work/09/593b381f4e3da5089ff973e0ac9448 + +Tip: you can try to figure out what's wrong by changing to the process work dir and showing the script file named `.command.sh` +May-02 15:59:23.548 [TaskFinalizer-2] INFO nextflow.Session - Execution cancelled -- Finishing pending tasks before exit +May-02 15:59:23.575 [TaskFinalizer-2] ERROR nextflow.Nextflow - Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting +May-02 15:59:23.577 [main] DEBUG nextflow.Session - Session await > all processes finished +May-02 15:59:23.599 [Actor Thread 18] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process +May-02 15:59:23.602 [Actor Thread 18] DEBUG nextflow.processor.TaskContext - Failed to serialize delegate map items: [ + 'meta':[Script_df773f5847ef0bc7$_runScript_closure1$_closure7] = + 'pos_range':[java.lang.String] = 352-1383 + '$':[java.lang.Boolean] = true + 'wt_seq':[nextflow.processor.TaskPath] = GID1A_SUNi_ref_small.fasta + 'script':[nextflow.processor.TaskPath] = aa_seq.R + 'task':[nextflow.processor.TaskConfig] = [container:community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure15@6c0e933f, withName:FASTQC:[ext:[args:--quiet], containerOptions:], withLabel:error_retry:[errorStrategy:retry, maxRetries:2], when:nextflow.script.TaskClosure@591d5648, withLabel:process_high:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure23@773dbcd3, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure24@2400957, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure25@1acff54b], withLabel:error_ignore:[errorStrategy:ignore], resourceLimits:[cpus:4, memory:8.GB, time:1.h], withLabel:process_high_memory:[memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure11$_closure27@29d34f4c], publishDir:[[path:Script7B16D34DB7A7F1FB6B799846EECD6610$_run_closure1$_closure2@3392a581, mode:copy, saveAs:Script7B16D34DB7A7F1FB6B799846EECD6610$_run_closure1$_closure3@319a1815]], executor:local, stub:nextflow.script.TaskClosure@549ad6c0, conda:null/environment.yml, cacheable:true, withLabel:process_low:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure17@67005a30, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure18@411845fc, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure19@20a3b77a], withLabel:process_medium:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure20@220d4978, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure21@47b956d3, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure22@448ddc7d], withName:MULTIQC:[ext:[args:Script7B16D34DB7A7F1FB6B799846EECD6610$_run_closure1$_closure5$_closure6@79a8bbf1], publishDir:[path:Script7B16D34DB7A7F1FB6B799846EECD6610$_run_closure1$_closure5$_closure7@31266b35, mode:copy, saveAs:Script7B16D34DB7A7F1FB6B799846EECD6610$_run_closure1$_closure5$_closure8@3c24ac0c]], workDir:/Users/benjaminwehnert/dmscore/work/09/593b381f4e3da5089ff973e0ac9448, exitStatus:1, errorCount:1, ext:[:], withLabel:process_single:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure14@16e0f1c1, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure15@6c0e933f, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure16@2a2ba7df], process:NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ, debug:false, cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure14@16e0f1c1, retryCount:1, index:1, label:[process_single], withLabel:process_long:[time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure10$_closure26@28561e6c], maxRetries:1, maxErrors:-1, shell:bash + +set -e # Exit if a tool returns a non-zero status/exit code +set -u # Treat unset variables and parameters as an error +set -o pipefail # Returns the status of the last command to exit with a non-zero status or zero if all successfully execute +set -C # No clobber - prevent output redirection from overwriting files. +, name:NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1), containerOptions:-u $(id -u):$(id -g), errorStrategy:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure5@310b93a6, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure16@2a2ba7df, hash:09593b381f4e3da5089ff973e0ac9448] +] +com.esotericsoftware.kryo.KryoException: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue +Serialization trace: +queue (org.codehaus.groovy.util.ReferenceManager$CallBackedManager) +manager (org.codehaus.groovy.util.ReferenceManager$1) +manager (org.codehaus.groovy.util.ReferenceBundle) +bundle (org.codehaus.groovy.reflection.CachedClass$4) +cachedSuperClass (org.codehaus.groovy.reflection.stdclasses.ObjectCachedClass) +cachedClass (org.codehaus.groovy.reflection.CachedMethod) +allMethods (groovy.lang.MetaClassImpl) +delegate (groovy.runtime.metaclass.NextflowDelegatingMetaClass) +metaClass (groovyx.gpars.dataflow.DataflowVariable) +first (groovyx.gpars.dataflow.stream.DataflowStream) +asyncHead (groovyx.gpars.dataflow.stream.DataflowStreamReadAdapter) +source (nextflow.extension.MapOp) +owner (nextflow.extension.MapOp$_apply_closure1) +code (groovyx.gpars.dataflow.operator.DataflowOperatorActor) +actor (groovyx.gpars.dataflow.operator.DataflowOperator) +allOperators (nextflow.Session) +session (nextflow.validation.ValidationExtension) +target (nextflow.script.FunctionDef) +definitions (nextflow.script.ScriptMeta) +meta (nextflow.script.ScriptBinding) +binding (Script_df773f5847ef0bc7) +delegate (Script_df773f5847ef0bc7$_runScript_closure1) +delegate (Script_df773f5847ef0bc7$_runScript_closure1$_closure7) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:82) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:82) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:22) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:82) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:22) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:95) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:21) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:95) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:21) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.util.KryoHelper.serialize(SerializationHelper.groovy:166) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.processor.TaskContext.serialize(TaskContext.groovy:198) + at nextflow.cache.CacheDB.writeTaskEntry0(CacheDB.groovy:148) + at java.base/jdk.internal.reflect.NativeMethodAccessorImpl.invoke0(Native Method) + at java.base/jdk.internal.reflect.NativeMethodAccessorImpl.invoke(NativeMethodAccessorImpl.java:77) + at java.base/jdk.internal.reflect.DelegatingMethodAccessorImpl.invoke(DelegatingMethodAccessorImpl.java:43) + at java.base/java.lang.reflect.Method.invoke(Method.java:569) + at org.codehaus.groovy.reflection.CachedMethod.invoke(CachedMethod.java:343) + at groovy.lang.MetaMethod.doMethodInvoke(MetaMethod.java:328) + at groovy.lang.MetaClassImpl.doInvokeMethod(MetaClassImpl.java:1333) + at groovy.lang.MetaClassImpl.invokeMethod(MetaClassImpl.java:1088) + at groovy.lang.MetaClassImpl.invokeMethod(MetaClassImpl.java:1007) + at org.codehaus.groovy.runtime.InvokerHelper.invokePogoMethod(InvokerHelper.java:645) + at org.codehaus.groovy.runtime.InvokerHelper.invokeMethod(InvokerHelper.java:628) + at org.codehaus.groovy.runtime.InvokerHelper.invokeMethodSafe(InvokerHelper.java:82) + at nextflow.cache.CacheDB$_putTaskAsync_closure1.doCall(CacheDB.groovy:157) + at nextflow.cache.CacheDB$_putTaskAsync_closure1.call(CacheDB.groovy) + at groovyx.gpars.agent.AgentBase.onMessage(AgentBase.java:102) + at groovyx.gpars.agent.Agent.handleMessage(Agent.java:84) + at groovyx.gpars.agent.AgentCore$1.handleMessage(AgentCore.java:48) + at groovyx.gpars.util.AsyncMessagingCore.run(AsyncMessagingCore.java:132) + at java.base/java.util.concurrent.ThreadPoolExecutor.runWorker(ThreadPoolExecutor.java:1136) + at java.base/java.util.concurrent.ThreadPoolExecutor$Worker.run(ThreadPoolExecutor.java:635) + at java.base/java.lang.Thread.run(Thread.java:840) +Caused by: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue + at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:48) + at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:26) + at com.esotericsoftware.kryo.Kryo.newDefaultSerializer(Kryo.java:351) + at com.esotericsoftware.kryo.Kryo.getDefaultSerializer(Kryo.java:344) + at com.esotericsoftware.kryo.util.DefaultClassResolver.registerImplicit(DefaultClassResolver.java:56) + at com.esotericsoftware.kryo.Kryo.getRegistration(Kryo.java:461) + at com.esotericsoftware.kryo.util.DefaultClassResolver.writeClass(DefaultClassResolver.java:79) + at com.esotericsoftware.kryo.Kryo.writeClass(Kryo.java:488) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:57) + ... 106 common frames omitted +Caused by: java.lang.reflect.InvocationTargetException: null + at jdk.internal.reflect.GeneratedConstructorAccessor37.newInstance(Unknown Source) + at java.base/jdk.internal.reflect.DelegatingConstructorAccessorImpl.newInstance(DelegatingConstructorAccessorImpl.java:45) + at java.base/java.lang.reflect.Constructor.newInstanceWithCaller(Constructor.java:500) + at java.base/java.lang.reflect.Constructor.newInstance(Constructor.java:481) + at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:35) + ... 114 common frames omitted +Caused by: java.lang.reflect.InaccessibleObjectException: Unable to make field private final java.lang.ref.ReferenceQueue$Lock java.lang.ref.ReferenceQueue.lock accessible: module java.base does not "opens java.lang.ref" to unnamed module @7b02881e + at java.base/java.lang.reflect.AccessibleObject.checkCanSetAccessible(AccessibleObject.java:354) + at java.base/java.lang.reflect.AccessibleObject.checkCanSetAccessible(AccessibleObject.java:297) + at java.base/java.lang.reflect.Field.checkCanSetAccessible(Field.java:178) + at java.base/java.lang.reflect.Field.setAccessible(Field.java:172) + at com.esotericsoftware.kryo.serializers.FieldSerializer.buildValidFields(FieldSerializer.java:282) + at com.esotericsoftware.kryo.serializers.FieldSerializer.rebuildCachedFields(FieldSerializer.java:217) + at com.esotericsoftware.kryo.serializers.FieldSerializer.rebuildCachedFields(FieldSerializer.java:156) + at com.esotericsoftware.kryo.serializers.FieldSerializer.(FieldSerializer.java:133) + ... 119 common frames omitted +May-02 15:59:33.713 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 4; name: NFCORE_DMSCORE:DMSCORE:MULTIQC; status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/0c/0721f606c722113c3be3676c55bf64] +May-02 15:59:33.768 [Task monitor] DEBUG n.processor.TaskPollingMonitor - <<< barrier arrives (monitor: local) - terminating tasks monitor poll loop +May-02 15:59:33.769 [main] DEBUG nextflow.Session - Session await > all barriers passed +May-02 15:59:34.181 [TaskFinalizer-3] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:MULTIQC > Skipping output binding because one or more optional files are missing: fileoutparam<2> +May-02 15:59:34.199 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'TaskFinalizer' shutdown completed (hard=false) +May-02 15:59:34.204 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'PublishDir' shutdown completed (hard=false) +May-02 15:59:34.218 [main] INFO nextflow.Nextflow - -[nf-core/dmscore] Pipeline completed with errors- +May-02 15:59:34.228 [main] DEBUG n.trace.WorkflowStatsObserver - Workflow completed > WorkflowStats[succeededCount=2; failedCount=1; ignoredCount=0; cachedCount=1; pendingCount=1; submittedCount=0; runningCount=0; retriesCount=0; abortedCount=0; succeedDuration=8.6s; failedDuration=1m 26s; cachedDuration=48s;loadCpus=0; loadMemory=0; peakRunning=1; peakCpus=1; peakMemory=6 GB; ] +May-02 15:59:34.229 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow completed -- saving trace file +May-02 15:59:34.233 [main] DEBUG nextflow.trace.ReportObserver - Workflow completed -- rendering execution report +May-02 15:59:35.624 [main] DEBUG nextflow.trace.TimelineObserver - Workflow completed -- rendering execution timeline +May-02 15:59:35.837 [main] DEBUG nextflow.cache.CacheDB - Closing CacheDB done +May-02 15:59:35.864 [main] INFO org.pf4j.AbstractPluginManager - Stop plugin 'nf-schema@2.3.0' +May-02 15:59:35.864 [main] DEBUG nextflow.plugin.BasePlugin - Plugin stopped nf-schema +May-02 15:59:35.865 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'FileTransfer' shutdown completed (hard=false) +May-02 15:59:35.867 [main] DEBUG nextflow.script.ScriptRunner - > Execution complete -- Goodbye diff --git a/modules/local/possiblemutations.nf b/modules/local/possiblemutations.nf new file mode 100644 index 0000000..4c44188 --- /dev/null +++ b/modules/local/possiblemutations.nf @@ -0,0 +1,89 @@ +// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) +// https://github.com/nf-core/modules/tree/master/modules/nf-core/ +// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: +// https://nf-co.re/join +// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. +// All other parameters MUST be provided using the "task.ext" directive, see here: +// https://www.nextflow.io/docs/latest/process.html#ext +// where "task.ext" is a string. +// Any parameters that need to be evaluated in the context of a particular sample +// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. +// TODO nf-core: Software that can be piped together SHOULD be added to separate module files +// unless there is a run-time, storage advantage in implementing in this way +// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: +// bwa mem | samtools view -B -T ref.fasta +// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty +// list (`[]`) instead of a file can be used to work around this issue. + +process POSSIBLEMUTATIONS { + tag '$bam' + label 'process_single' + + // TODO nf-core: List required Conda package(s). + // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). + // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. + // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/YOUR-TOOL-HERE': + 'biocontainers/YOUR-TOOL-HERE' }" + + input: + // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" + // MUST be provided as an input via a Groovy Map called "meta". + // This information may not be required in some instances e.g. indexing reference genome files: + // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf + // TODO nf-core: Where applicable please provide/convert compressed files as input/output + // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. + path bam + + output: + // TODO nf-core: Named file extensions MUST be emitted for ALL output channels + path "*.bam", emit: bam + // TODO nf-core: List additional required output channels/values here + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + + // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 + // If the software is unable to output a version number on the command-line then it can be manually specified + // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf + // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) + // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive + // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter + // using the Nextflow "task" variable e.g. "--threads $task.cpus" + // TODO nf-core: Please replace the example samtools command below with your module's command + // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) + """ + samtools \\ + sort \\ + $args \\ + -@ $task.cpus \\ + $bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + possiblemutations: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ + + stub: + def args = task.ext.args ?: '' + + // TODO nf-core: A stub section should mimic the execution of the original module as best as possible + // Have a look at the following examples: + // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 + // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 + """ + touch ${prefix}.bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + possiblemutations: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ +} From 15c422fa1e32638554d5351c6d5db5ae38993965 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Tue, 6 May 2025 16:53:29 +0200 Subject: [PATCH 095/317] move possible mutations script --- modules/local/{ => dmsanalysis}/possiblemutations.nf | 0 1 file changed, 0 insertions(+), 0 deletions(-) rename modules/local/{ => dmsanalysis}/possiblemutations.nf (100%) diff --git a/modules/local/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf similarity index 100% rename from modules/local/possiblemutations.nf rename to modules/local/dmsanalysis/possiblemutations.nf From bf0f2932172b4f6284b2303165950815d48a42c6 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 6 May 2025 17:08:01 +0200 Subject: [PATCH 096/317] Update possiblemutations.nf --- .../local/dmsanalysis/possiblemutations.nf | 88 ++++--------------- 1 file changed, 19 insertions(+), 69 deletions(-) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index 4c44188..66af6c3 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -1,89 +1,39 @@ -// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) -// https://github.com/nf-core/modules/tree/master/modules/nf-core/ -// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: -// https://nf-co.re/join -// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. -// All other parameters MUST be provided using the "task.ext" directive, see here: -// https://www.nextflow.io/docs/latest/process.html#ext -// where "task.ext" is a string. -// Any parameters that need to be evaluated in the context of a particular sample -// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. -// TODO nf-core: Software that can be piped together SHOULD be added to separate module files -// unless there is a run-time, storage advantage in implementing in this way -// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: -// bwa mem | samtools view -B -T ref.fasta -// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty -// list (`[]`) instead of a file can be used to work around this issue. - -process POSSIBLEMUTATIONS { - tag '$bam' +process DMSANALYSIS_POSSIBLE_MUTATIONS { label 'process_single' - // TODO nf-core: List required Conda package(s). - // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). - // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. - // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/YOUR-TOOL-HERE': - 'biocontainers/YOUR-TOOL-HERE' }" + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" input: - // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" - // MUST be provided as an input via a Groovy Map called "meta". - // This information may not be required in some instances e.g. indexing reference genome files: - // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf - // TODO nf-core: Where applicable please provide/convert compressed files as input/output - // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. - path bam + path wt_seq + val pos_range + val mutagenesis_type + path custom_codon_library + path script // possible_mutations.R output: - // TODO nf-core: Named file extensions MUST be emitted for ALL output channels - path "*.bam", emit: bam - // TODO nf-core: List additional required output channels/values here - path "versions.yml" , emit: versions + path "possible_mutations.csv", emit: possible_mutations + path "versions.yml", emit: versions when: task.ext.when == null || task.ext.when script: - def args = task.ext.args ?: '' - - // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 - // If the software is unable to output a version number on the command-line then it can be manually specified - // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf - // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) - // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive - // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter - // using the Nextflow "task" variable e.g. "--threads $task.cpus" - // TODO nf-core: Please replace the example samtools command below with your module's command - // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) """ - samtools \\ - sort \\ - $args \\ - -@ $task.cpus \\ - $bam + start_stop_codon="$pos_range" + + if [[ "$custom_codon_library" == "/NULL" ]]; then + Rscript -e "source('$script'); generate_possible_variants('$wt_seq', '\$start_stop_codon', '$mutagenesis_type', NULL, 'possible_mutations.csv')" + else + Rscript -e "source('$script'); generate_possible_variants('$wt_seq', '\$start_stop_codon', '$mutagenesis_type', '$custom_codon_library', 'possible_mutations.csv')" + fi - cat <<-END_VERSIONS > versions.yml - "${task.process}": - possiblemutations: \$(samtools --version |& sed '1!d ; s/samtools //') - END_VERSIONS + echo "." > versions.yml """ stub: - def args = task.ext.args ?: '' - - // TODO nf-core: A stub section should mimic the execution of the original module as best as possible - // Have a look at the following examples: - // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 - // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 """ - touch ${prefix}.bam - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - possiblemutations: \$(samtools --version |& sed '1!d ; s/samtools //') - END_VERSIONS + touch possible_mutations.csv + echo "stub" > versions.yml """ } From 599f5b177866f4e90a3561a318d8e11f1e3c9617 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 6 May 2025 17:19:45 +0200 Subject: [PATCH 097/317] Update dmscore.nf --- workflows/dmscore.nf | 28 ++++++++++++++++++++++++++-- 1 file changed, 26 insertions(+), 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index bc8379b..ea8bf70 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -9,8 +9,9 @@ include { BWA_INDEX } from '../modules/nf-core/bwa/index/main' include { BWA_MEM } from '../modules/nf-core/bwa/mem/main' include { BAMFILTER_DMS } from '../modules/local/bamfilteringdms' include { PREMERGE } from '../modules/local/premerge' -include { GATK_SATURATIONMUTAGENESIS } from '../modules/local/gatk/saturationmutagenesis' +include { GATK_SATURATIONMUTAGENESIS } from '../modules/local/gatk/saturationmutagenesis' include { DMSANALYSIS_AASEQ } from '../modules/local/dmsanalysis/aaseq' +include { DMSANALYSIS_POSSIBLE_MUTATIONS } from '../modules/local/dmsanalysis/possiblemutations' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -22,6 +23,12 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ +// Params defaults +params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library + + // Define fasta file as channel (e.g. for BWA index) Channel .fromPath(params.fasta, checkIfExists: true) @@ -34,11 +41,20 @@ Channel .set { reading_frame_ch } // Define min_counts as channel (e.g. for gatk function) -> if not set - default: 3 -params.min_counts = params.min_counts ?: 3 Channel .value(params.min_counts) .set { min_counts_ch } +// Define custom library as channel +Channel + .value(params.custom_codon_library) + .set { custom_codon_library_ch } + +// Define mutagenesis type as channel +Channel + .value(params.mutagenesis_type) + .set { mutagenesis_type_ch } + // Define R scripts as channels Channel.fromPath("modules/local/dmsanalysis/bin/SeqDepth_simulation.R", checkIfExists: true).set { seqdepth_simulation_script_ch } Channel.fromPath("modules/local/dmsanalysis/bin/aa_seq.R", checkIfExists: true).set { aa_seq_script_ch } @@ -171,6 +187,14 @@ workflow DMSCORE { aa_seq_script_ch // path to aa_seq.R (defined at the top) ) + DMSANALYSIS_POSSIBLE_MUTATIONS( + ch_fasta.map{ it[1] }, // wt_seq (as path) + reading_frame_ch, // pos_range (as val) + mutagenesis_type_ch, // mutagenesis_type (as val) + custom_codon_library_ch, // custom_codon_library (as path) + possible_mutations_script_ch // path to R script + ) + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From 18d9883cfbcd839e00642c9313035021662870ae Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 6 May 2025 17:26:44 +0200 Subject: [PATCH 098/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index 66af6c3..6644e8a 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -1,4 +1,5 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { + tag "table /w all possible variants" label 'process_single' conda "${moduleDir}/environment.yml" From 548dd7d086ff7290f17c92fa62d14c5929e3d3f7 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Tue, 6 May 2025 17:44:17 +0200 Subject: [PATCH 099/317] restructure folders --- modules/local/{ => bamprocessing}/bamfilteringdms.nf | 0 modules/local/{ => bamprocessing}/premerge.nf | 0 modules/local/{dmsanalysis => gatk}/annotategatk.nf | 0 modules/local/{dmsanalysis => visualization}/visualization.nf | 0 4 files changed, 0 insertions(+), 0 deletions(-) rename modules/local/{ => bamprocessing}/bamfilteringdms.nf (100%) rename modules/local/{ => bamprocessing}/premerge.nf (100%) rename modules/local/{dmsanalysis => gatk}/annotategatk.nf (100%) rename modules/local/{dmsanalysis => visualization}/visualization.nf (100%) diff --git a/modules/local/bamfilteringdms.nf b/modules/local/bamprocessing/bamfilteringdms.nf similarity index 100% rename from modules/local/bamfilteringdms.nf rename to modules/local/bamprocessing/bamfilteringdms.nf diff --git a/modules/local/premerge.nf b/modules/local/bamprocessing/premerge.nf similarity index 100% rename from modules/local/premerge.nf rename to modules/local/bamprocessing/premerge.nf diff --git a/modules/local/dmsanalysis/annotategatk.nf b/modules/local/gatk/annotategatk.nf similarity index 100% rename from modules/local/dmsanalysis/annotategatk.nf rename to modules/local/gatk/annotategatk.nf diff --git a/modules/local/dmsanalysis/visualization.nf b/modules/local/visualization/visualization.nf similarity index 100% rename from modules/local/dmsanalysis/visualization.nf rename to modules/local/visualization/visualization.nf From b4aa379c949f8bfe2094f03be66acae8130cf073 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Tue, 6 May 2025 17:45:39 +0200 Subject: [PATCH 100/317] Update dmscore.nf --- workflows/dmscore.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index ea8bf70..f70c809 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -7,8 +7,8 @@ include { FASTQC } from '../modules/nf-core/fastqc/main' include { MULTIQC } from '../modules/nf-core/multiqc/main' include { BWA_INDEX } from '../modules/nf-core/bwa/index/main' include { BWA_MEM } from '../modules/nf-core/bwa/mem/main' -include { BAMFILTER_DMS } from '../modules/local/bamfilteringdms' -include { PREMERGE } from '../modules/local/premerge' +include { BAMFILTER_DMS } from '../modules/local/bamprocessing/bamfilteringdms' +include { PREMERGE } from '../modules/local/bamprocessing/premerge' include { GATK_SATURATIONMUTAGENESIS } from '../modules/local/gatk/saturationmutagenesis' include { DMSANALYSIS_AASEQ } from '../modules/local/dmsanalysis/aaseq' include { DMSANALYSIS_POSSIBLE_MUTATIONS } from '../modules/local/dmsanalysis/possiblemutations' From 7c201fedfbaef8844aa8937a57a49006da9dbd82 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 09:39:50 +0200 Subject: [PATCH 101/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 2 ++ 1 file changed, 2 insertions(+) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index 6644e8a..8f5e55b 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -5,6 +5,8 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + publishDir "${params.outdir}/intermediate_files", mode: 'copy' + input: path wt_seq val pos_range From fd9406a1ac20ed03d968fe9d9c28ce4e48436a20 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:10:53 +0200 Subject: [PATCH 102/317] Update dmscore.nf --- workflows/dmscore.nf | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index f70c809..599be2d 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -76,6 +76,11 @@ Channel.fromPath("modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_he Channel.fromPath("modules/local/dmsanalysis/bin/process_raw_gatk.R", checkIfExists: true).set { process_raw_gatk_script_ch } +// Centralized output paths +output_intermediate_files = "${params.outdir}/intermediate_files" +output_gatk = "${params.outdir}/intermediate_files/gatk" +output_final_plots = "${params.outdir}/final_plots" +output_processed_gatk = "${params.outdir}/intermediate_files/processed_gatk_files" workflow DMSCORE { From cf99cafb1fd9a56488c28c08dd373b149e0f5196 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:11:29 +0200 Subject: [PATCH 103/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index 8f5e55b..5df3d87 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -5,7 +5,7 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" - publishDir "${params.outdir}/intermediate_files", mode: 'copy' + publishDir "${output_intermediate_files}", mode: 'copy' input: path wt_seq From 13a8e645475b25f3e628b72bd0ec3b295990c1ba Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:38:34 +0200 Subject: [PATCH 104/317] Update dmscore.nf --- workflows/dmscore.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 599be2d..945a560 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -24,10 +24,10 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc */ // Params defaults -params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library - +params.outdir = params.outdir ?: './results' // default output folder is "results" inside the wd // Define fasta file as channel (e.g. for BWA index) Channel From 17f9a6c02b44f52a5da5ca59104b5d6160e0f37f Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:41:24 +0200 Subject: [PATCH 105/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index 5df3d87..f9da708 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -5,7 +5,7 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" - publishDir "${output_intermediate_files}", mode: 'copy' + publishDir "${params.output_intermediate_files}", mode: 'copy' input: path wt_seq From 8419034bb5bfb17e77ec9f0305b2b51300c86793 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:42:16 +0200 Subject: [PATCH 106/317] Update dmscore.nf --- workflows/dmscore.nf | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 945a560..51e6a90 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -77,10 +77,10 @@ Channel.fromPath("modules/local/dmsanalysis/bin/process_raw_gatk.R", checkIfExis // Centralized output paths -output_intermediate_files = "${params.outdir}/intermediate_files" -output_gatk = "${params.outdir}/intermediate_files/gatk" -output_final_plots = "${params.outdir}/final_plots" -output_processed_gatk = "${params.outdir}/intermediate_files/processed_gatk_files" +params.output_intermediate_files = "${params.outdir}/intermediate_files" +params.output_gatk = "${params.outdir}/intermediate_files/gatk" +params.output_final_plots = "${params.outdir}/final_plots" +params.output_processed_gatk = "${params.outdir}/intermediate_files/processed_gatk_files" workflow DMSCORE { From 043384cebc078086cd45be47497b0f4f283fd07a Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:43:48 +0200 Subject: [PATCH 107/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index f9da708..baac46a 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -5,7 +5,7 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" - publishDir "${params.output_intermediate_files}", mode: 'copy' + publishDir params.output_intermediate_files, mode: 'copy' input: path wt_seq From 2c0d4a3b9fbb6ae547272d6e1eedb84a0c5bd4d1 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:46:22 +0200 Subject: [PATCH 108/317] Update dmscore.nf --- workflows/dmscore.nf | 5 +++-- 1 file changed, 3 insertions(+), 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 51e6a90..8c32a3c 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -27,7 +27,7 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library -params.outdir = params.outdir ?: './results' // default output folder is "results" inside the wd +params.outdir = params.outdir ?: "./results" // default output folder is "results" inside the wd // Define fasta file as channel (e.g. for BWA index) Channel @@ -77,12 +77,13 @@ Channel.fromPath("modules/local/dmsanalysis/bin/process_raw_gatk.R", checkIfExis // Centralized output paths -params.output_intermediate_files = "${params.outdir}/intermediate_files" +params.output_intermediate_files = params.output_intermediate_files ?: "${params.outdir}/intermediate_files" params.output_gatk = "${params.outdir}/intermediate_files/gatk" params.output_final_plots = "${params.outdir}/final_plots" params.output_processed_gatk = "${params.outdir}/intermediate_files/processed_gatk_files" + workflow DMSCORE { take: From 60023178078cc3469f868e13f7d1ba09ec9d4304 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:47:25 +0200 Subject: [PATCH 109/317] Update dmscore.nf --- workflows/dmscore.nf | 8 -------- 1 file changed, 8 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 8c32a3c..d2c6442 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -76,14 +76,6 @@ Channel.fromPath("modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_he Channel.fromPath("modules/local/dmsanalysis/bin/process_raw_gatk.R", checkIfExists: true).set { process_raw_gatk_script_ch } -// Centralized output paths -params.output_intermediate_files = params.output_intermediate_files ?: "${params.outdir}/intermediate_files" -params.output_gatk = "${params.outdir}/intermediate_files/gatk" -params.output_final_plots = "${params.outdir}/final_plots" -params.output_processed_gatk = "${params.outdir}/intermediate_files/processed_gatk_files" - - - workflow DMSCORE { take: From 89fcc694c4259aede8bd1ecd7ae0973274670398 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:48:16 +0200 Subject: [PATCH 110/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index baac46a..8f5e55b 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -5,7 +5,7 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" - publishDir params.output_intermediate_files, mode: 'copy' + publishDir "${params.outdir}/intermediate_files", mode: 'copy' input: path wt_seq From 273cbb622ccf75da00de871cbc5776db70933463 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:50:23 +0200 Subject: [PATCH 111/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index d2c6442..825120e 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -27,7 +27,7 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library -params.outdir = params.outdir ?: "./results" // default output folder is "results" inside the wd +params.outdir = params.outdir ?: './results' // default output folder is "results" inside the wd // Define fasta file as channel (e.g. for BWA index) Channel From 34a0853180f5e3f207f0377d7da819b8e4836f4e Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:51:29 +0200 Subject: [PATCH 112/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 825120e..939824b 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -27,7 +27,7 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library -params.outdir = params.outdir ?: './results' // default output folder is "results" inside the wd +params.outdir = params.outdir ?: 'results' // default output folder is "results" inside the wd // Define fasta file as channel (e.g. for BWA index) Channel From 9041601a4c7dbd40b65bd89712959c096a0d0c10 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:52:09 +0200 Subject: [PATCH 113/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 939824b..4f2c6d0 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -27,7 +27,7 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library -params.outdir = params.outdir ?: 'results' // default output folder is "results" inside the wd +params.outdir = params.outdir ?: "results" // default output folder is "results" inside the wd // Define fasta file as channel (e.g. for BWA index) Channel From 298ff18c257953cdacb8a2fad003b7bb50251823 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:52:41 +0200 Subject: [PATCH 114/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 4f2c6d0..345cc70 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -27,7 +27,7 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library -params.outdir = params.outdir ?: "results" // default output folder is "results" inside the wd +params.outdir = "results" // default output folder is "results" inside the wd // Define fasta file as channel (e.g. for BWA index) Channel From 5adbeb42f0b7d1da11cfb94218cc4d916fb2922c Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:53:24 +0200 Subject: [PATCH 115/317] Update dmscore.nf --- workflows/dmscore.nf | 1 - 1 file changed, 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 345cc70..4c1e1a0 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -27,7 +27,6 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library -params.outdir = "results" // default output folder is "results" inside the wd // Define fasta file as channel (e.g. for BWA index) Channel From f55191da420f608e7c1f46b0c89417735ecd2805 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:54:30 +0200 Subject: [PATCH 116/317] Update dmscore.nf --- workflows/dmscore.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 4c1e1a0..eef055e 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -27,6 +27,7 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library +params.outdir = params.outdir ?: ./results // Define fasta file as channel (e.g. for BWA index) Channel From 0dc0891dd9a4ee0ee43839b098e95e2ccd6d227b Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 10:54:58 +0200 Subject: [PATCH 117/317] Update dmscore.nf --- workflows/dmscore.nf | 1 - 1 file changed, 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index eef055e..4c1e1a0 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -27,7 +27,6 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library -params.outdir = params.outdir ?: ./results // Define fasta file as channel (e.g. for BWA index) Channel From 4d81bac61d4c752f250b487554d3759cb5822891 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 11:03:19 +0200 Subject: [PATCH 118/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 8 ++++++-- 1 file changed, 6 insertions(+), 2 deletions(-) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index 8f5e55b..442f48a 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -31,12 +31,16 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { Rscript -e "source('$script'); generate_possible_variants('$wt_seq', '\$start_stop_codon', '$mutagenesis_type', '$custom_codon_library', 'possible_mutations.csv')" fi - echo "." > versions.yml + # Capture R base and packages versions + echo "DMSANALYSIS_POSSIBLE_MUTATIONS:" > versions.yml + echo " r-base: \$(R --version | head -n 1 | sed 's/^R version //')" >> versions.yml + echo " biostrings: \$(Rscript -e 'packageVersion(\"Biostrings\")' | tail -n 1 | sed 's/\\[1\\] //')" >> versions.yml """ stub: """ touch possible_mutations.csv - echo "stub" > versions.yml + echo "DMSANALYSIS_POSSIBLE_MUTATIONS:" > versions.yml + echo " stub-version: 0.0.0" >> versions.yml """ } From 0f2d9051e0c0f05b614723e69188502acde0e939 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 11:09:28 +0200 Subject: [PATCH 119/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 12 ++++++++---- 1 file changed, 8 insertions(+), 4 deletions(-) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index 442f48a..7172bdf 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -31,10 +31,14 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { Rscript -e "source('$script'); generate_possible_variants('$wt_seq', '\$start_stop_codon', '$mutagenesis_type', '$custom_codon_library', 'possible_mutations.csv')" fi - # Capture R base and packages versions - echo "DMSANALYSIS_POSSIBLE_MUTATIONS:" > versions.yml - echo " r-base: \$(R --version | head -n 1 | sed 's/^R version //')" >> versions.yml - echo " biostrings: \$(Rscript -e 'packageVersion(\"Biostrings\")' | tail -n 1 | sed 's/\\[1\\] //')" >> versions.yml + # Extract R base and packages versions + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + BIOSTRINGS_VERSION=\$(Rscript -e "packageVersion('Biostrings')" | grep -Eo '[0-9]+(\\.[0-9]+)+') + cat <<-END_VERSIONS > versions.yml + DMSANALYSIS_POSSIBLE_MUTATIONS: + r-base: \$R_VERSION + biostrings: \$BIOSTRINGS_VERSION + END_VERSIONS """ stub: From 431278b50c0c802213ff4c546e3a4b9f8c5233d7 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 11:15:42 +0200 Subject: [PATCH 120/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 20 +++++++++++--------- 1 file changed, 11 insertions(+), 9 deletions(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index fc17294..d90f79f 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -5,6 +5,8 @@ process DMSANALYSIS_AASEQ { conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + publishDir "${params.outdir}/intermediate_files", mode: 'copy' + input: tuple val(meta), path(wt_seq) val pos_range @@ -25,20 +27,20 @@ process DMSANALYSIS_AASEQ { Rscript -e "source('$script'); aa_seq('$wt_seq', '\$start_stop_codon', 'aa_seq.txt')" + # Extract R base and packages versions + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + BIOSTRINGS_VERSION=\$(Rscript -e "packageVersion('Biostrings')" | grep -Eo '[0-9]+(\\.[0-9]+)+') cat <<-END_VERSIONS > versions.yml - "DMSANALYSIS_AASEQ": - r: \$R_version - aa_seq_script: custom_R_script_bin - END_VERSIONS + DMSANALYSIS_AASEQ: + r-base: \$R_VERSION + biostrings: \$BIOSTRINGS_VERSION + END_VERSIONS """ stub: """ touch aa_seq.txt - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - dmsanalysis_aaseq: dummy_version - END_VERSIONS + echo "DMSANALYSIS_AASEQ:" > versions.yml + echo " stub-version: 0.0.0" >> versions.yml """ } From 4047a3c893ea0f2e419bbc5a42b860da88206d51 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 11:22:20 +0200 Subject: [PATCH 121/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index 7172bdf..08181e6 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -8,14 +8,14 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { publishDir "${params.outdir}/intermediate_files", mode: 'copy' input: - path wt_seq + tuple val(meta), path(wt_seq) val pos_range val mutagenesis_type path custom_codon_library path script // possible_mutations.R output: - path "possible_mutations.csv", emit: possible_mutations + tuple val(meta), path("possible_mutations.csv"), emit: possible_mutations path "versions.yml", emit: versions when: From ddf66e1c3185fc415c99596e45a01bedc698c9d9 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 11:22:53 +0200 Subject: [PATCH 122/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 4c1e1a0..7b05f59 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -185,7 +185,7 @@ workflow DMSCORE { ) DMSANALYSIS_POSSIBLE_MUTATIONS( - ch_fasta.map{ it[1] }, // wt_seq (as path) + ch_fasta reading_frame_ch, // pos_range (as val) mutagenesis_type_ch, // mutagenesis_type (as val) custom_codon_library_ch, // custom_codon_library (as path) From cd918756cffb2f6605248c55d491ecfc1a95c129 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 11:25:33 +0200 Subject: [PATCH 123/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 7b05f59..1245cc4 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -185,7 +185,7 @@ workflow DMSCORE { ) DMSANALYSIS_POSSIBLE_MUTATIONS( - ch_fasta + ch_fasta, reading_frame_ch, // pos_range (as val) mutagenesis_type_ch, // mutagenesis_type (as val) custom_codon_library_ch, // custom_codon_library (as path) From c1f464147bec44d0fc5ca488dfc169b1941c3527 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 11:35:10 +0200 Subject: [PATCH 124/317] Update dmscore.nf --- workflows/dmscore.nf | 3 +++ 1 file changed, 3 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 1245cc4..6a4e85d 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -192,6 +192,9 @@ workflow DMSCORE { possible_mutations_script_ch // path to R script ) + ch_versions = ch_versions.mix(DMSANALYSIS_POSSIBLE_MUTATIONS.out.versions) + + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From 7326ed1db2dc99d7c5cdfbdee0b651c01c15d4a2 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 11:54:16 +0200 Subject: [PATCH 125/317] Update nextflow_schema.json --- nextflow_schema.json | 10 ++++++++++ 1 file changed, 10 insertions(+) diff --git a/nextflow_schema.json b/nextflow_schema.json index 74cdacc..0e40b84 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -52,7 +52,17 @@ "description": "Minimum number of variant observations required.", "minimum": 1, "default": 3 + }, + "reading_frame": { + "type": "string", + "description": "Type of mutagenic primers. Choose from nnk, nns, max_diff_to_wt, custom. When using 'custom', also provide the parameter 'custom_codon_library'", + "default": "nnk" } + "custom_codon_library": { + "type": "string", + "format": "file-path", + "description": "Path to a comma-separated .txt file listing custom codons (e.g., 'AAA,AAC,AAG,...'). Required when mutagenesis_type is 'custom'." + } } }, "reference_genome_options": { From ae9928ca17bc42b79b20d93b97e2b1489086f7e1 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 12:59:06 +0200 Subject: [PATCH 126/317] Update nextflow_schema.json --- nextflow_schema.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 0e40b84..049fccb 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -57,7 +57,7 @@ "type": "string", "description": "Type of mutagenic primers. Choose from nnk, nns, max_diff_to_wt, custom. When using 'custom', also provide the parameter 'custom_codon_library'", "default": "nnk" - } + }, "custom_codon_library": { "type": "string", "format": "file-path", From 2dc910ae2589960ee1aad8c87c6d1eb14e0072ed Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 13:00:39 +0200 Subject: [PATCH 127/317] Update nextflow_schema.json --- nextflow_schema.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 049fccb..cc08d2f 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -53,7 +53,7 @@ "minimum": 1, "default": 3 }, - "reading_frame": { + "mutagenesis_type": { "type": "string", "description": "Type of mutagenic primers. Choose from nnk, nns, max_diff_to_wt, custom. When using 'custom', also provide the parameter 'custom_codon_library'", "default": "nnk" From 04a19f3c3d77addf02e4de97cfdf2fe7a728626b Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 15:32:38 +0200 Subject: [PATCH 128/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 6a4e85d..3de6fe1 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -183,6 +183,7 @@ workflow DMSCORE { reading_frame_ch, aa_seq_script_ch // path to aa_seq.R (defined at the top) ) + ch_versions = ch_versions.mix(DMSANALYSIS_AASEQ.out.versions) DMSANALYSIS_POSSIBLE_MUTATIONS( ch_fasta, @@ -191,7 +192,6 @@ workflow DMSCORE { custom_codon_library_ch, // custom_codon_library (as path) possible_mutations_script_ch // path to R script ) - ch_versions = ch_versions.mix(DMSANALYSIS_POSSIBLE_MUTATIONS.out.versions) From 1800f68408f7188ccae5f2e976c6741af7459c9f Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 21:22:35 +0200 Subject: [PATCH 129/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 2 -- 1 file changed, 2 deletions(-) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index 08181e6..ea7d9cf 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -5,8 +5,6 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" - publishDir "${params.outdir}/intermediate_files", mode: 'copy' - input: tuple val(meta), path(wt_seq) val pos_range From 5eb97d1af465ebdfbcc6191659706e43f1b021dd Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 21:22:53 +0200 Subject: [PATCH 130/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 2 -- 1 file changed, 2 deletions(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index d90f79f..ec45ec1 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -5,8 +5,6 @@ process DMSANALYSIS_AASEQ { conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" - publishDir "${params.outdir}/intermediate_files", mode: 'copy' - input: tuple val(meta), path(wt_seq) val pos_range From 1438b13e9a86c04ca369c4d4c04c486beb8cf60d Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 21:24:57 +0200 Subject: [PATCH 131/317] Update modules.config --- conf/modules.config | 6 ++++++ 1 file changed, 6 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index 36edb1d..e07e4a1 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -32,4 +32,10 @@ process { ] } + process { + withName:DMSANALYSIS_POSSIBLE_MUTATIONS { + publishDir = [ "${params.outdir}/intermediate_files", mode: 'copy' ] + } + } + } From 7e428ac91357b48175d8c6ba4ee1736c2a4ae1f7 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 21:26:20 +0200 Subject: [PATCH 132/317] Update modules.config --- conf/modules.config | 4 +--- 1 file changed, 1 insertion(+), 3 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index e07e4a1..21e02e5 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -32,10 +32,8 @@ process { ] } - process { - withName:DMSANALYSIS_POSSIBLE_MUTATIONS { + withName:DMSANALYSIS_POSSIBLE_MUTATIONS { publishDir = [ "${params.outdir}/intermediate_files", mode: 'copy' ] - } } } From 0dec1e42e75e42a623347e6a1fa64ba11a9a0d65 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 21:28:07 +0200 Subject: [PATCH 133/317] Update modules.config --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index 21e02e5..51ec599 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -32,7 +32,7 @@ process { ] } - withName:DMSANALYSIS_POSSIBLE_MUTATIONS { + withName: DMSANALYSIS_POSSIBLE_MUTATIONS { publishDir = [ "${params.outdir}/intermediate_files", mode: 'copy' ] } From e7bd7bc07528ce1eb84d6966c8d4e1f4a113af8b Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 21:29:00 +0200 Subject: [PATCH 134/317] Update modules.config --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index 51ec599..d76e22f 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -32,7 +32,7 @@ process { ] } - withName: DMSANALYSIS_POSSIBLE_MUTATIONS { + withName: 'DMSANALYSIS_POSSIBLE_MUTATIONS' { publishDir = [ "${params.outdir}/intermediate_files", mode: 'copy' ] } From 413085d458d7165c581d6a96eec7dad7463fe508 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 21:34:14 +0200 Subject: [PATCH 135/317] Update modules.config --- conf/modules.config | 6 +++++- 1 file changed, 5 insertions(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index d76e22f..a7805da 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -33,7 +33,11 @@ process { } withName: 'DMSANALYSIS_POSSIBLE_MUTATIONS' { - publishDir = [ "${params.outdir}/intermediate_files", mode: 'copy' ] + publishDir = [ + path: "${params.outdir}/intermediate_files", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] } } From 0cda7331cdfd31a98706a7fc6171671da68fa84f Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 21:46:00 +0200 Subject: [PATCH 136/317] Update modules.config --- conf/modules.config | 24 ++++++++++++++++++++++++ 1 file changed, 24 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index a7805da..93a1a25 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -32,6 +32,22 @@ process { ] } + withName: 'BWA_INDEX' { + publishDir = [ + path: "${params.outdir}/intermediate_files/bwa/index", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + + withName: 'BWA_MEM' { + publishDir = [ + path: "${params.outdir}/intermediate_files/bwa/mem", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + withName: 'DMSANALYSIS_POSSIBLE_MUTATIONS' { publishDir = [ path: "${params.outdir}/intermediate_files", @@ -40,4 +56,12 @@ process { ] } + withName: 'DMSANALYSIS_AASEQ' { + publishDir = [ + path: "${params.outdir}/intermediate_files", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + } From 4d7ccd01b638d902aa6ee4d4adbae0149efd7f5a Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 21:56:31 +0200 Subject: [PATCH 137/317] Update modules.config --- conf/modules.config | 36 ++++++++++++++++++++++++++++++++++-- 1 file changed, 34 insertions(+), 2 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 93a1a25..a7da483 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -34,7 +34,7 @@ process { withName: 'BWA_INDEX' { publishDir = [ - path: "${params.outdir}/intermediate_files/bwa/index", + path: "${params.outdir}/intermediate_files/bam_files", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] @@ -42,7 +42,31 @@ process { withName: 'BWA_MEM' { publishDir = [ - path: "${params.outdir}/intermediate_files/bwa/mem", + path: "${params.outdir}/intermediate_files/bam_files/bwa/mem", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + + withName: 'BAMFILTER_DMS' { + publishDir = [ + path: "${params.outdir}/intermediate_files/bam_files", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + + withName: 'PREMERGE' { + publishDir = [ + path: "${params.outdir}/intermediate_files/bam_files", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + + withName: 'GATK_SATURATIONMUTAGENESIS' { + publishDir = [ + path: "${params.outdir}/intermediate_files/gatk", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] @@ -64,4 +88,12 @@ process { ] } + withName: 'DMSANALYSIS_AASEQ' { + publishDir = [ + path: "${params.outdir}/intermediate_files", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + } From 25525a0fe228087e383c93c37016ca1d6b97ee85 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Wed, 7 May 2025 22:18:43 +0200 Subject: [PATCH 138/317] process gatk script --- log.txt | 534 +++++------------------ modules/local/dmsanalysis/processgatk.nf | 91 ++++ 2 files changed, 206 insertions(+), 419 deletions(-) create mode 100644 modules/local/dmsanalysis/processgatk.nf diff --git a/log.txt b/log.txt index c55d577..4511556 100644 --- a/log.txt +++ b/log.txt @@ -1,428 +1,124 @@ -May-02 15:57:46.796 [main] DEBUG nextflow.cli.Launcher - $> nextflow run . -profile test,docker --fasta /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta --reading_frame 352-1383 --min_counts 2 --outdir ./results -resume -May-02 15:57:46.924 [main] DEBUG nextflow.cli.CmdRun - N E X T F L O W ~ version 24.04.4 -May-02 15:57:46.948 [main] DEBUG nextflow.plugin.PluginsFacade - Setting up plugin manager > mode=prod; embedded=false; plugins-dir=/Users/benjaminwehnert/.nextflow/plugins; core-plugins: nf-amazon@2.5.3,nf-azure@1.6.1,nf-cloudcache@0.4.1,nf-codecommit@0.2.1,nf-console@1.1.3,nf-ga4gh@1.3.0,nf-google@1.13.2-patch1,nf-tower@1.9.1,nf-wave@1.4.2-patch1 -May-02 15:57:46.960 [main] INFO o.pf4j.DefaultPluginStatusProvider - Enabled plugins: [] -May-02 15:57:46.961 [main] INFO o.pf4j.DefaultPluginStatusProvider - Disabled plugins: [] -May-02 15:57:46.963 [main] INFO org.pf4j.DefaultPluginManager - PF4J version 3.12.0 in 'deployment' mode -May-02 15:57:46.979 [main] INFO org.pf4j.AbstractPluginManager - No plugins -May-02 15:57:47.853 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` -May-02 15:57:47.871 [main] DEBUG nextflow.config.ConfigBuilder - Found config local: /Users/benjaminwehnert/dmscore/nextflow.config -May-02 15:57:47.879 [main] DEBUG nextflow.config.ConfigBuilder - Parsing config file: /Users/benjaminwehnert/dmscore/nextflow.config -May-02 15:57:47.897 [main] DEBUG n.secret.LocalSecretsProvider - Secrets store: /Users/benjaminwehnert/.nextflow/secrets/store.json -May-02 15:57:47.900 [main] DEBUG nextflow.secret.SecretsLoader - Discovered secrets providers: [nextflow.secret.LocalSecretsProvider@7ee8130e] - activable => nextflow.secret.LocalSecretsProvider@7ee8130e -May-02 15:57:47.910 [main] DEBUG nextflow.config.ConfigBuilder - Applying config profile: `test,docker` -May-02 15:57:49.363 [main] DEBUG nextflow.config.ConfigBuilder - Available config profiles: [bih, cfc_dev, uzl_omics, ifb_core, embl_hd, denbi_qbic, alice, mjolnir_globe, uppmax, giga, incliva, ilifu, ki_luria, uge, icr_alma, rosalind_uge, lugh, mccleary, unibe_ibu, vai, czbiohub_aws, jax, roslin, ccga_med, tes, scw, unc_longleaf, tigem, tubingen_apg, google, apollo, ipop_up, vsc_calcua, pdc_kth, googlels, ceci_nic5, humantechnopole, stjude, daisybio, eddie, medair, biowulf, apptainer, bi, bigpurple, adcra, cedars, pawsey_setonix, vsc_kul_uhasselt, pawsey_nimbus, ucl_myriad, utd_ganymede, charliecloud, seattlechildrens, icr_davros, ceres, arm, munin, rosalind, hasta, cfc, uzh, shu_bmrc, ebi_codon_slurm, ebc, ccga_dx, crick, ku_sund_danhead, marvin, shifter, biohpc_gen, mana, mamba, york_viking, unc_lccc, wehi, awsbatch, wustl_htcf, arcc, ceci_dragon2, imperial, maestro, software_license, cannon, genotoul, nci_gadi, abims, janelia, nu_genomics, googlebatch, oist, sahmri, kaust, alliance_canada, mpcdf, leicester, vsc_ugent, create, sage, cambridge, jex, podman, ebi_codon, cheaha, xanadu, nyu_hpc, test, marjorie, computerome, ucd_sonic, seg_globe, sanger, dkfz, bluebear, pasteur, einstein, ethz_euler, m3c, test_full, imb, ucl_cscluster, tuos_stanage, azurebatch, hki, seadragon, crukmi, csiro_petrichor, qmul_apocrita, wave, docker, engaging, gis, hypatia, psmn, eva, unity, cropdiversityhpc, nygc, fgcz, conda, crg, singularity, mpcdf_viper, pe2, self_hosted_runner, tufts, uw_hyak_pedslabs, binac2, debug, genouest, cbe, unsw_katana, gitpod, phoenix, seawulf, uod_hpc, fub_curta, uct_hpc, aws_tower, binac, fsu_draco] -May-02 15:57:49.415 [main] DEBUG nextflow.cli.CmdRun - Applied DSL=2 by global default -May-02 15:57:49.433 [main] DEBUG nextflow.cli.CmdRun - Launching `./main.nf` [golden_curran] DSL2 - revision: 84101fc51c -May-02 15:57:49.435 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins declared=[nf-schema@2.3.0] -May-02 15:57:49.435 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins default=[] -May-02 15:57:49.436 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins resolved requirement=[nf-schema@2.3.0] -May-02 15:57:49.436 [main] DEBUG nextflow.plugin.PluginUpdater - Installing plugin nf-schema version: 2.3.0 -May-02 15:57:49.444 [main] INFO org.pf4j.AbstractPluginManager - Plugin 'nf-schema@2.3.0' resolved -May-02 15:57:49.444 [main] INFO org.pf4j.AbstractPluginManager - Start plugin 'nf-schema@2.3.0' -May-02 15:57:49.450 [main] DEBUG nextflow.plugin.BasePlugin - Plugin started nf-schema@2.3.0 -May-02 15:57:49.513 [main] DEBUG nextflow.Session - Session UUID: 36458275-f468-409a-bb18-98996facae84 -May-02 15:57:49.513 [main] DEBUG nextflow.Session - Run name: golden_curran -May-02 15:57:49.513 [main] DEBUG nextflow.Session - Executor pool size: 8 -May-02 15:57:49.521 [main] DEBUG nextflow.file.FilePorter - File porter settings maxRetries=3; maxTransfers=50; pollTimeout=null -May-02 15:57:49.531 [main] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'FileTransfer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false -May-02 15:57:49.553 [main] DEBUG nextflow.cli.CmdRun - - Version: 24.04.4 build 5917 - Created: 01-08-2024 07:05 UTC (09:05 CEST) - System: Mac OS X 15.0 - Runtime: Groovy 4.0.21 on OpenJDK 64-Bit Server VM 17.0.13+0 - Encoding: UTF-8 (UTF-8) - Process: 36172@MacBook-Air-von-Benjamin.local [127.0.0.1] - CPUs: 8 - Mem: 8 GB (162.4 MB) - Swap: 8 GB (449.1 MB) -May-02 15:57:49.575 [main] DEBUG nextflow.Session - Work-dir: /Users/benjaminwehnert/dmscore/work [Mac OS X] -May-02 15:57:49.575 [main] DEBUG nextflow.Session - Script base path does not exist or is not a directory: /Users/benjaminwehnert/dmscore/bin -May-02 15:57:49.591 [main] DEBUG nextflow.executor.ExecutorFactory - Extension executors providers=[] -May-02 15:57:49.599 [main] DEBUG nextflow.Session - Observer factory: DefaultObserverFactory -May-02 15:57:49.619 [main] DEBUG nextflow.Session - Observer factory: ValidationObserverFactory -May-02 15:57:49.650 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` -May-02 15:57:49.675 [main] DEBUG nextflow.cache.CacheFactory - Using Nextflow cache factory: nextflow.cache.DefaultCacheFactory -May-02 15:57:49.685 [main] DEBUG nextflow.util.CustomThreadPool - Creating default thread pool > poolSize: 9; maxThreads: 1000 -May-02 15:57:49.783 [main] DEBUG nextflow.Session - Session start -May-02 15:57:49.785 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow started -- trace file: /Users/benjaminwehnert/dmscore/results/pipeline_info/execution_trace_2025-05-02_15-57-48.txt -May-02 15:57:49.944 [main] DEBUG nextflow.script.ScriptRunner - > Launching execution -May-02 15:57:51.084 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema -May-02 15:57:51.555 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema -May-02 15:57:51.556 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema -May-02 15:57:51.559 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema -May-02 15:57:51.560 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema -May-02 15:57:52.021 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema -May-02 15:57:52.022 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema -May-02 15:57:52.024 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema -May-02 15:57:52.025 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema -May-02 15:57:52.227 [main] INFO nextflow.Nextflow - ------------------------------------------------------- - ,--./,-. - ___ __ __ __ ___ /,-._.--~' - |\ | |__ __ / ` / \ |__) |__ } { - | \| | \__, \__/ | \ |___ \`-._,-`-, - `._,._,' - nf-core/dmscore 1.0.0dev ------------------------------------------------------- -Input/output options - input : https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv - outdir : ./results - min_counts : 2 - -Reference genome options - genome : R64-1-1 - fasta : /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta - -Institutional config options - config_profile_name : Test profile - config_profile_description: Minimal test dataset to check pipeline function - -Generic options - trace_report_suffix : 2025-05-02_15-57-48 - -Core Nextflow options - runName : golden_curran - containerEngine : docker - launchDir : /Users/benjaminwehnert/dmscore - workDir : /Users/benjaminwehnert/dmscore/work - projectDir : /Users/benjaminwehnert/dmscore - userName : benjaminwehnert - profile : test,docker - configFiles : /Users/benjaminwehnert/dmscore/nextflow.config - -!! Only displaying parameters that differ from the pipeline defaults !! ------------------------------------------------------- -* The nf-core framework - https://doi.org/10.1038/s41587-020-0439-x - -* Software dependencies - https://github.com/nf-core/dmscore/blob/master/CITATIONS.md - -May-02 15:57:52.229 [main] DEBUG n.validation.ValidationExtension - Starting parameters validation -May-02 15:57:52.604 [main] DEBUG nextflow.validation.SchemaEvaluator - Started validating /BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv -May-02 15:57:53.970 [main] DEBUG nextflow.validation.SchemaEvaluator - Validation of file 'https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv' passed! -May-02 15:57:54.378 [main] DEBUG n.v.FormatDirectoryPathEvaluator - Cloud blob storage paths are not supported by 'FormatDirectoryPathEvaluator': 's3://ngi-igenomes/igenomes/' -May-02 15:57:54.380 [main] DEBUG n.validation.ValidationExtension - Finishing parameters validation -May-02 15:57:54.462 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:FASTQC -May-02 15:57:54.464 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:FASTQC` matches process NFCORE_DMSCORE:DMSCORE:FASTQC -May-02 15:57:54.477 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -May-02 15:57:54.477 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -May-02 15:57:54.483 [main] DEBUG nextflow.executor.Executor - [warm up] executor > local -May-02 15:57:54.488 [main] DEBUG n.processor.LocalPollingMonitor - Creating local task monitor for executor 'local' > cpus=8; memory=8 GB; capacity=8; pollInterval=100ms; dumpInterval=5m -May-02 15:57:54.490 [main] DEBUG n.processor.TaskPollingMonitor - >>> barrier register (monitor: local) -May-02 15:57:54.672 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:MULTIQC -May-02 15:57:54.673 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:MULTIQC` matches process NFCORE_DMSCORE:DMSCORE:MULTIQC -May-02 15:57:54.676 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -May-02 15:57:54.676 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -May-02 15:57:54.686 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BWA_INDEX -May-02 15:57:54.688 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -May-02 15:57:54.688 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -May-02 15:57:54.702 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_high` matches labels `process_high` for process with name NFCORE_DMSCORE:DMSCORE:BWA_MEM -May-02 15:57:54.706 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -May-02 15:57:54.706 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -May-02 15:57:54.715 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS -May-02 15:57:54.718 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -May-02 15:57:54.718 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -May-02 15:57:54.729 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:PREMERGE -May-02 15:57:54.732 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -May-02 15:57:54.732 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -May-02 15:57:54.746 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_high` matches labels `process_high` for process with name NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS -May-02 15:57:54.750 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -May-02 15:57:54.750 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -May-02 15:57:54.761 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ -May-02 15:57:54.764 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null -May-02 15:57:54.764 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' -May-02 15:57:54.781 [main] DEBUG nextflow.Session - Config process names validation disabled as requested -May-02 15:57:54.783 [main] DEBUG nextflow.Session - Igniting dataflow network (22) -May-02 15:57:54.790 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:FASTQC -May-02 15:57:54.792 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:MULTIQC -May-02 15:57:54.793 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX -May-02 15:57:54.794 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_MEM -May-02 15:57:54.794 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS -May-02 15:57:54.794 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:PREMERGE -May-02 15:57:54.795 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS -May-02 15:57:54.795 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ -May-02 15:57:54.797 [main] DEBUG nextflow.script.ScriptRunner - Parsed script files: - Script_e5f965e09aa3641b: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/premerge.nf - Script_113f045ba19c2c46: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/fastqc/main.nf - Script_4f612cfd8c52e8cd: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nextflow_pipeline/main.nf - Script_215a636da7ab24a2: /Users/benjaminwehnert/dmscore/./subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfschema_plugin/main.nf - Script_68606fccdecc54d9: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf - Script_f88958acfb50d604: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/dmsanalysis/aaseq.nf - Script_df773f5847ef0bc7: /Users/benjaminwehnert/dmscore/./workflows/dmscore.nf - Script_0f3077e98f6a93f6: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/mem/main.nf - Script_5c4e8d4051efa81e: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/nf-core/utils_nfcore_pipeline/main.nf - Script_4ff7366a79e0ef06: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/bamfilteringdms.nf - Script_c568aae5b239c1c5: /Users/benjaminwehnert/dmscore/main.nf - Script_4955288afd8ca61e: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/multiqc/main.nf - Script_37dcd664b2773148: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/gatk/saturationmutagenesis.nf - Script_25aa31c18d513c61: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/index/main.nf -May-02 15:57:54.797 [main] DEBUG nextflow.script.ScriptRunner - > Awaiting termination -May-02 15:57:54.798 [main] DEBUG nextflow.Session - Session await -May-02 15:57:54.893 [Actor Thread 10] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.002 s; external sort time: 0.016 s; total time: 0.018 s -May-02 15:57:54.893 [Actor Thread 1] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.002 s; external sort time: 0.016 s; total time: 0.018 s -May-02 15:57:54.894 [Actor Thread 1] DEBUG nextflow.file.FileCollector - >> temp file exists? false -May-02 15:57:54.894 [Actor Thread 10] DEBUG nextflow.file.FileCollector - >> temp file exists? false -May-02 15:57:54.895 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Missed collect-file cache -- cause: java.nio.file.NoSuchFileException: /Users/benjaminwehnert/dmscore/work/collect-file/9aa80ec1e5fc421feabac9f320c60e5e -May-02 15:57:54.895 [Actor Thread 10] DEBUG nextflow.file.FileCollector - Missed collect-file cache -- cause: java.nio.file.NoSuchFileException: /Users/benjaminwehnert/dmscore/work/collect-file/a9ac31ffb91d381d31c639634831f9b7 -May-02 15:57:54.901 [Actor Thread 10] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/a9ac31ffb91d381d31c639634831f9b7 -May-02 15:57:54.901 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/9aa80ec1e5fc421feabac9f320c60e5e -May-02 15:57:54.910 [Actor Thread 10] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-1468229241367644128 -May-02 15:57:54.910 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-9598669176738179442 -May-02 15:57:54.920 [Actor Thread 15] DEBUG nextflow.util.HashBuilder - [WARN] Unknown hashing type: class Script_df773f5847ef0bc7$_runScript_closure1$_closure7 -May-02 15:57:55.001 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -May-02 15:57:55.003 [Task submitter] INFO nextflow.Session - [a4/7e1d06] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta) -May-02 15:57:55.734 [FileTransfer-2] DEBUG nextflow.file.FilePorter - Local cache found for foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq at /Users/benjaminwehnert/dmscore/work/stage-36458275-f468-409a-bb18-98996facae84/cc/f938a8775ac8befd4337b2732c7dc9/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq -May-02 15:57:55.734 [FileTransfer-1] DEBUG nextflow.file.FilePorter - Local cache found for foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq at /Users/benjaminwehnert/dmscore/work/stage-36458275-f468-409a-bb18-98996facae84/28/abb932c7e69700944b1605c1ee72d2/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq -May-02 15:57:55.867 [Actor Thread 9] INFO nextflow.processor.TaskProcessor - [5a/035f68] Cached process > NFCORE_DMSCORE:DMSCORE:FASTQC (gid1a_1_quality_1_pe) -May-02 15:57:55.967 [Actor Thread 9] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'PublishDir' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false -May-02 15:57:56.063 [Actor Thread 18] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 2; slices: 1; internal sort time: 0.022 s; external sort time: 0.002 s; total time: 0.024 s -May-02 15:57:56.063 [Actor Thread 18] DEBUG nextflow.file.FileCollector - >> temp file exists? true -May-02 15:57:56.065 [Actor Thread 18] DEBUG nextflow.file.FileCollector - Retrieved cached collect-files from: /Users/benjaminwehnert/dmscore/work/collect-file/0dde6dd9bfe7393330209153ab0f204f -May-02 15:57:56.070 [Actor Thread 18] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-5944054719096968617 -May-02 15:57:57.568 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 1; name: NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/a4/7e1d06298bb5191af0791289427bba] -May-02 15:57:57.569 [Task monitor] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'TaskFinalizer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false -May-02 15:57:57.621 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -May-02 15:57:57.623 [Task submitter] INFO nextflow.Session - [09/593b38] Submitted process > NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1) -May-02 15:57:57.692 [Actor Thread 17] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process -May-02 15:59:23.448 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 2; name: NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1); status: COMPLETED; exit: 1; error: -; workDir: /Users/benjaminwehnert/dmscore/work/09/593b381f4e3da5089ff973e0ac9448] -May-02 15:59:23.464 [TaskFinalizer-2] DEBUG nextflow.processor.TaskProcessor - Handling unexpected condition for - task: name=NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1); work-dir=/Users/benjaminwehnert/dmscore/work/09/593b381f4e3da5089ff973e0ac9448 - error [nextflow.exception.ProcessFailedException]: Process `NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1)` terminated with an error exit status (1) -May-02 15:59:23.479 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run -May-02 15:59:23.480 [Task submitter] INFO nextflow.Session - [0c/0721f6] Submitted process > NFCORE_DMSCORE:DMSCORE:MULTIQC -May-02 15:59:23.536 [TaskFinalizer-2] ERROR nextflow.processor.TaskProcessor - Error executing process > 'NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1)' - -Caused by: - Process `NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1)` terminated with an error exit status (1) - - -Command executed: - - start_stop_codon="352-1383" - - Rscript -e "source('aa_seq.R'); aa_seq('GID1A_SUNi_ref_small.fasta', '$start_stop_codon', 'aa_seq.txt')" +May-07 21:29:10.689 [main] DEBUG nextflow.cli.Launcher - $> nextflow run . -profile test,docker --fasta /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta --reading_frame 352-1383 --min_counts 2 --mutagenesis_type max_diff_to_wt --outdir ./results +May-07 21:29:10.864 [main] DEBUG nextflow.cli.CmdRun - N E X T F L O W ~ version 24.04.4 +May-07 21:29:10.887 [main] DEBUG nextflow.plugin.PluginsFacade - Setting up plugin manager > mode=prod; embedded=false; plugins-dir=/Users/benjaminwehnert/.nextflow/plugins; core-plugins: nf-amazon@2.5.3,nf-azure@1.6.1,nf-cloudcache@0.4.1,nf-codecommit@0.2.1,nf-console@1.1.3,nf-ga4gh@1.3.0,nf-google@1.13.2-patch1,nf-tower@1.9.1,nf-wave@1.4.2-patch1 +May-07 21:29:10.897 [main] INFO o.pf4j.DefaultPluginStatusProvider - Enabled plugins: [] +May-07 21:29:10.897 [main] INFO o.pf4j.DefaultPluginStatusProvider - Disabled plugins: [] +May-07 21:29:10.900 [main] INFO org.pf4j.DefaultPluginManager - PF4J version 3.12.0 in 'deployment' mode +May-07 21:29:10.914 [main] INFO org.pf4j.AbstractPluginManager - No plugins +May-07 21:29:11.769 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` +May-07 21:29:11.790 [main] DEBUG nextflow.config.ConfigBuilder - Found config local: /Users/benjaminwehnert/dmscore/nextflow.config +May-07 21:29:11.798 [main] DEBUG nextflow.config.ConfigBuilder - Parsing config file: /Users/benjaminwehnert/dmscore/nextflow.config +May-07 21:29:11.815 [main] DEBUG n.secret.LocalSecretsProvider - Secrets store: /Users/benjaminwehnert/.nextflow/secrets/store.json +May-07 21:29:11.818 [main] DEBUG nextflow.secret.SecretsLoader - Discovered secrets providers: [nextflow.secret.LocalSecretsProvider@6296474f] - activable => nextflow.secret.LocalSecretsProvider@6296474f +May-07 21:29:11.830 [main] DEBUG nextflow.config.ConfigBuilder - Applying config profile: `test,docker` +May-07 21:29:13.163 [main] ERROR nextflow.cli.Launcher - Unable to parse config file: '/Users/benjaminwehnert/dmscore/nextflow.config' + + Compile failed for sources FixedSetSources[name='/groovy/script/Script7BD980DA7F8768B9E9654BF7A071C3F7']. Cause: org.codehaus.groovy.control.MultipleCompilationErrorsException: startup failed: + /groovy/script/Script7BD980DA7F8768B9E9654BF7A071C3F7: 13: Unexpected input: '{' @ line 13, column 9. + process { + ^ - cat <<-END_VERSIONS > versions.yml - "${task.process}": - r: $(R --version | head -n 1 | sed 's/^R version //') - aa_seq_script: custom_R_script_bin - END_VERSIONS - -Command exit status: - 1 - -Command output: - (empty) - -Command error: - Unable to find image 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' locally - 0fd2e39a5bf2ecaa: Pulling from library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned - 6114410fdbce: Pulling fs layer - bc6461884445: Pulling fs layer - bc6461884445: Download complete - 6114410fdbce: Download complete - Digest: sha256:2e209ed7eb4e3c8c5f29a6e9f65dc2113c00e69b5c7dc4ff5bc5abbd2b080242 - Status: Downloaded newer image for community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa - .command.sh: line 12: "${task.process}": - r: $(R --version | head -n 1 | sed 's/^R version //') - aa_seq_script: custom_R_script_bin - : bad substitution - -Work dir: - /Users/benjaminwehnert/dmscore/work/09/593b381f4e3da5089ff973e0ac9448 - -Tip: you can try to figure out what's wrong by changing to the process work dir and showing the script file named `.command.sh` -May-02 15:59:23.548 [TaskFinalizer-2] INFO nextflow.Session - Execution cancelled -- Finishing pending tasks before exit -May-02 15:59:23.575 [TaskFinalizer-2] ERROR nextflow.Nextflow - Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting -May-02 15:59:23.577 [main] DEBUG nextflow.Session - Session await > all processes finished -May-02 15:59:23.599 [Actor Thread 18] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process -May-02 15:59:23.602 [Actor Thread 18] DEBUG nextflow.processor.TaskContext - Failed to serialize delegate map items: [ - 'meta':[Script_df773f5847ef0bc7$_runScript_closure1$_closure7] = - 'pos_range':[java.lang.String] = 352-1383 - '$':[java.lang.Boolean] = true - 'wt_seq':[nextflow.processor.TaskPath] = GID1A_SUNi_ref_small.fasta - 'script':[nextflow.processor.TaskPath] = aa_seq.R - 'task':[nextflow.processor.TaskConfig] = [container:community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure15@6c0e933f, withName:FASTQC:[ext:[args:--quiet], containerOptions:], withLabel:error_retry:[errorStrategy:retry, maxRetries:2], when:nextflow.script.TaskClosure@591d5648, withLabel:process_high:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure23@773dbcd3, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure24@2400957, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure25@1acff54b], withLabel:error_ignore:[errorStrategy:ignore], resourceLimits:[cpus:4, memory:8.GB, time:1.h], withLabel:process_high_memory:[memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure11$_closure27@29d34f4c], publishDir:[[path:Script7B16D34DB7A7F1FB6B799846EECD6610$_run_closure1$_closure2@3392a581, mode:copy, saveAs:Script7B16D34DB7A7F1FB6B799846EECD6610$_run_closure1$_closure3@319a1815]], executor:local, stub:nextflow.script.TaskClosure@549ad6c0, conda:null/environment.yml, cacheable:true, withLabel:process_low:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure17@67005a30, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure18@411845fc, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure19@20a3b77a], withLabel:process_medium:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure20@220d4978, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure21@47b956d3, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure22@448ddc7d], withName:MULTIQC:[ext:[args:Script7B16D34DB7A7F1FB6B799846EECD6610$_run_closure1$_closure5$_closure6@79a8bbf1], publishDir:[path:Script7B16D34DB7A7F1FB6B799846EECD6610$_run_closure1$_closure5$_closure7@31266b35, mode:copy, saveAs:Script7B16D34DB7A7F1FB6B799846EECD6610$_run_closure1$_closure5$_closure8@3c24ac0c]], workDir:/Users/benjaminwehnert/dmscore/work/09/593b381f4e3da5089ff973e0ac9448, exitStatus:1, errorCount:1, ext:[:], withLabel:process_single:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure14@16e0f1c1, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure15@6c0e933f, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure16@2a2ba7df], process:NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ, debug:false, cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure14@16e0f1c1, retryCount:1, index:1, label:[process_single], withLabel:process_long:[time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure10$_closure26@28561e6c], maxRetries:1, maxErrors:-1, shell:bash - -set -e # Exit if a tool returns a non-zero status/exit code -set -u # Treat unset variables and parameters as an error -set -o pipefail # Returns the status of the last command to exit with a non-zero status or zero if all successfully execute -set -C # No clobber - prevent output redirection from overwriting files. -, name:NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (1), containerOptions:-u $(id -u):$(id -g), errorStrategy:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure5@310b93a6, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure16@2a2ba7df, hash:09593b381f4e3da5089ff973e0ac9448] -] -com.esotericsoftware.kryo.KryoException: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -Serialization trace: -queue (org.codehaus.groovy.util.ReferenceManager$CallBackedManager) -manager (org.codehaus.groovy.util.ReferenceManager$1) -manager (org.codehaus.groovy.util.ReferenceBundle) -bundle (org.codehaus.groovy.reflection.CachedClass$4) -cachedSuperClass (org.codehaus.groovy.reflection.stdclasses.ObjectCachedClass) -cachedClass (org.codehaus.groovy.reflection.CachedMethod) -allMethods (groovy.lang.MetaClassImpl) -delegate (groovy.runtime.metaclass.NextflowDelegatingMetaClass) -metaClass (groovyx.gpars.dataflow.DataflowVariable) -first (groovyx.gpars.dataflow.stream.DataflowStream) -asyncHead (groovyx.gpars.dataflow.stream.DataflowStreamReadAdapter) -source (nextflow.extension.MapOp) -owner (nextflow.extension.MapOp$_apply_closure1) -code (groovyx.gpars.dataflow.operator.DataflowOperatorActor) -actor (groovyx.gpars.dataflow.operator.DataflowOperator) -allOperators (nextflow.Session) -session (nextflow.validation.ValidationExtension) -target (nextflow.script.FunctionDef) -definitions (nextflow.script.ScriptMeta) -meta (nextflow.script.ScriptBinding) -binding (Script_df773f5847ef0bc7) -delegate (Script_df773f5847ef0bc7$_runScript_closure1) -delegate (Script_df773f5847ef0bc7$_runScript_closure1$_closure7) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:82) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) - at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:82) - at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:22) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) - at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:82) - at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:22) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) - at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:95) - at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:21) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) - at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) - at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) - at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:95) - at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:21) - at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + 1 error + +ch.artecat.grengine.except.CompileException: Compile failed for sources FixedSetSources[name='/groovy/script/Script7BD980DA7F8768B9E9654BF7A071C3F7']. Cause: org.codehaus.groovy.control.MultipleCompilationErrorsException: startup failed: +/groovy/script/Script7BD980DA7F8768B9E9654BF7A071C3F7: 13: Unexpected input: '{' @ line 13, column 9. + process { + ^ + +1 error + + at ch.artecat.grengine.code.groovy.DefaultGroovyCompiler.compile(DefaultGroovyCompiler.java:260) + at ch.artecat.grengine.load.DefaultTopCodeCache.getUpToDateCode(DefaultTopCodeCache.java:80) + at ch.artecat.grengine.load.LayeredClassLoader.getTopLoader(LayeredClassLoader.java:251) + at ch.artecat.grengine.load.LayeredClassLoader.loadMainClass(LayeredClassLoader.java:202) + at ch.artecat.grengine.engine.LayeredEngine.loadMainClass(LayeredEngine.java:198) + at ch.artecat.grengine.Grengine.loadMainClass(Grengine.java:679) + at ch.artecat.grengine.BaseGrengine.load(BaseGrengine.java:538) + at ch.artecat.grengine.BaseGrengine.load(BaseGrengine.java:387) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.selectMethod(IndyInterface.java:355) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.config.ConfigBase.includeConfig(ConfigBase.groovy:112) + at java.base/jdk.internal.reflect.NativeMethodAccessorImpl.invoke0(Native Method) + at java.base/jdk.internal.reflect.NativeMethodAccessorImpl.invoke(NativeMethodAccessorImpl.java:77) + at java.base/jdk.internal.reflect.DelegatingMethodAccessorImpl.invoke(DelegatingMethodAccessorImpl.java:43) + at java.base/java.lang.reflect.Method.invoke(Method.java:569) + at org.codehaus.groovy.reflection.CachedMethod.invoke(CachedMethod.java:343) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.selectMethod(IndyInterface.java:355) at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.util.KryoHelper.serialize(SerializationHelper.groovy:166) + at nextflow.config.ConfigParser$_parse_closure5.doCall(ConfigParser.groovy:457) + at jdk.internal.reflect.GeneratedMethodAccessor19.invoke(Unknown Source) + at java.base/jdk.internal.reflect.DelegatingMethodAccessorImpl.invoke(DelegatingMethodAccessorImpl.java:43) + at java.base/java.lang.reflect.Method.invoke(Method.java:569) + at org.codehaus.groovy.reflection.CachedMethod.invoke(CachedMethod.java:343) + at org.codehaus.groovy.runtime.metaclass.ClosureMetaMethod.invoke(ClosureMetaMethod.java:88) + at groovy.lang.ExpandoMetaClass.invokeMethod(ExpandoMetaClass.java:1140) + at groovy.lang.MetaClassImpl.invokeMethod(MetaClassImpl.java:1007) + at groovy.lang.DelegatingMetaClass.invokeMethod(DelegatingMetaClass.java:165) at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.processor.TaskContext.serialize(TaskContext.groovy:198) - at nextflow.cache.CacheDB.writeTaskEntry0(CacheDB.groovy:148) + at _nf_config_af5e21ee.run(_nf_config_af5e21ee:288) at java.base/jdk.internal.reflect.NativeMethodAccessorImpl.invoke0(Native Method) at java.base/jdk.internal.reflect.NativeMethodAccessorImpl.invoke(NativeMethodAccessorImpl.java:77) at java.base/jdk.internal.reflect.DelegatingMethodAccessorImpl.invoke(DelegatingMethodAccessorImpl.java:43) at java.base/java.lang.reflect.Method.invoke(Method.java:569) at org.codehaus.groovy.reflection.CachedMethod.invoke(CachedMethod.java:343) - at groovy.lang.MetaMethod.doMethodInvoke(MetaMethod.java:328) - at groovy.lang.MetaClassImpl.doInvokeMethod(MetaClassImpl.java:1333) - at groovy.lang.MetaClassImpl.invokeMethod(MetaClassImpl.java:1088) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.config.ConfigParser$_parse_closure5.doCall(ConfigParser.groovy:457) + at jdk.internal.reflect.GeneratedMethodAccessor19.invoke(Unknown Source) + at java.base/jdk.internal.reflect.DelegatingMethodAccessorImpl.invoke(DelegatingMethodAccessorImpl.java:43) + at java.base/java.lang.reflect.Method.invoke(Method.java:569) + at org.codehaus.groovy.reflection.CachedMethod.invoke(CachedMethod.java:343) + at org.codehaus.groovy.runtime.metaclass.ClosureMetaMethod.invoke(ClosureMetaMethod.java:88) + at groovy.lang.ExpandoMetaClass.invokeMethod(ExpandoMetaClass.java:1140) at groovy.lang.MetaClassImpl.invokeMethod(MetaClassImpl.java:1007) - at org.codehaus.groovy.runtime.InvokerHelper.invokePogoMethod(InvokerHelper.java:645) - at org.codehaus.groovy.runtime.InvokerHelper.invokeMethod(InvokerHelper.java:628) - at org.codehaus.groovy.runtime.InvokerHelper.invokeMethodSafe(InvokerHelper.java:82) - at nextflow.cache.CacheDB$_putTaskAsync_closure1.doCall(CacheDB.groovy:157) - at nextflow.cache.CacheDB$_putTaskAsync_closure1.call(CacheDB.groovy) - at groovyx.gpars.agent.AgentBase.onMessage(AgentBase.java:102) - at groovyx.gpars.agent.Agent.handleMessage(Agent.java:84) - at groovyx.gpars.agent.AgentCore$1.handleMessage(AgentCore.java:48) - at groovyx.gpars.util.AsyncMessagingCore.run(AsyncMessagingCore.java:132) - at java.base/java.util.concurrent.ThreadPoolExecutor.runWorker(ThreadPoolExecutor.java:1136) - at java.base/java.util.concurrent.ThreadPoolExecutor$Worker.run(ThreadPoolExecutor.java:635) - at java.base/java.lang.Thread.run(Thread.java:840) -Caused by: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue - at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:48) - at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:26) - at com.esotericsoftware.kryo.Kryo.newDefaultSerializer(Kryo.java:351) - at com.esotericsoftware.kryo.Kryo.getDefaultSerializer(Kryo.java:344) - at com.esotericsoftware.kryo.util.DefaultClassResolver.registerImplicit(DefaultClassResolver.java:56) - at com.esotericsoftware.kryo.Kryo.getRegistration(Kryo.java:461) - at com.esotericsoftware.kryo.util.DefaultClassResolver.writeClass(DefaultClassResolver.java:79) - at com.esotericsoftware.kryo.Kryo.writeClass(Kryo.java:488) - at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:57) - ... 106 common frames omitted -Caused by: java.lang.reflect.InvocationTargetException: null - at jdk.internal.reflect.GeneratedConstructorAccessor37.newInstance(Unknown Source) - at java.base/jdk.internal.reflect.DelegatingConstructorAccessorImpl.newInstance(DelegatingConstructorAccessorImpl.java:45) - at java.base/java.lang.reflect.Constructor.newInstanceWithCaller(Constructor.java:500) - at java.base/java.lang.reflect.Constructor.newInstance(Constructor.java:481) - at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:35) - ... 114 common frames omitted -Caused by: java.lang.reflect.InaccessibleObjectException: Unable to make field private final java.lang.ref.ReferenceQueue$Lock java.lang.ref.ReferenceQueue.lock accessible: module java.base does not "opens java.lang.ref" to unnamed module @7b02881e - at java.base/java.lang.reflect.AccessibleObject.checkCanSetAccessible(AccessibleObject.java:354) - at java.base/java.lang.reflect.AccessibleObject.checkCanSetAccessible(AccessibleObject.java:297) - at java.base/java.lang.reflect.Field.checkCanSetAccessible(Field.java:178) - at java.base/java.lang.reflect.Field.setAccessible(Field.java:172) - at com.esotericsoftware.kryo.serializers.FieldSerializer.buildValidFields(FieldSerializer.java:282) - at com.esotericsoftware.kryo.serializers.FieldSerializer.rebuildCachedFields(FieldSerializer.java:217) - at com.esotericsoftware.kryo.serializers.FieldSerializer.rebuildCachedFields(FieldSerializer.java:156) - at com.esotericsoftware.kryo.serializers.FieldSerializer.(FieldSerializer.java:133) - ... 119 common frames omitted -May-02 15:59:33.713 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 4; name: NFCORE_DMSCORE:DMSCORE:MULTIQC; status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/0c/0721f606c722113c3be3676c55bf64] -May-02 15:59:33.768 [Task monitor] DEBUG n.processor.TaskPollingMonitor - <<< barrier arrives (monitor: local) - terminating tasks monitor poll loop -May-02 15:59:33.769 [main] DEBUG nextflow.Session - Session await > all barriers passed -May-02 15:59:34.181 [TaskFinalizer-3] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:MULTIQC > Skipping output binding because one or more optional files are missing: fileoutparam<2> -May-02 15:59:34.199 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'TaskFinalizer' shutdown completed (hard=false) -May-02 15:59:34.204 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'PublishDir' shutdown completed (hard=false) -May-02 15:59:34.218 [main] INFO nextflow.Nextflow - -[nf-core/dmscore] Pipeline completed with errors- -May-02 15:59:34.228 [main] DEBUG n.trace.WorkflowStatsObserver - Workflow completed > WorkflowStats[succeededCount=2; failedCount=1; ignoredCount=0; cachedCount=1; pendingCount=1; submittedCount=0; runningCount=0; retriesCount=0; abortedCount=0; succeedDuration=8.6s; failedDuration=1m 26s; cachedDuration=48s;loadCpus=0; loadMemory=0; peakRunning=1; peakCpus=1; peakMemory=6 GB; ] -May-02 15:59:34.229 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow completed -- saving trace file -May-02 15:59:34.233 [main] DEBUG nextflow.trace.ReportObserver - Workflow completed -- rendering execution report -May-02 15:59:35.624 [main] DEBUG nextflow.trace.TimelineObserver - Workflow completed -- rendering execution timeline -May-02 15:59:35.837 [main] DEBUG nextflow.cache.CacheDB - Closing CacheDB done -May-02 15:59:35.864 [main] INFO org.pf4j.AbstractPluginManager - Stop plugin 'nf-schema@2.3.0' -May-02 15:59:35.864 [main] DEBUG nextflow.plugin.BasePlugin - Plugin stopped nf-schema -May-02 15:59:35.865 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'FileTransfer' shutdown completed (hard=false) -May-02 15:59:35.867 [main] DEBUG nextflow.script.ScriptRunner - > Execution complete -- Goodbye + at groovy.lang.DelegatingMetaClass.invokeMethod(DelegatingMetaClass.java:165) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.selectMethod(IndyInterface.java:355) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.config.ConfigParser.parse(ConfigParser.groovy:486) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.config.ConfigParser.parse(ConfigParser.groovy:312) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.config.ConfigBuilder.parse0(ConfigBuilder.groovy:439) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.config.ConfigBuilder.merge0(ConfigBuilder.groovy:425) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.config.ConfigBuilder.buildConfig0(ConfigBuilder.groovy:375) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.config.ConfigBuilder.buildGivenFiles(ConfigBuilder.groovy:320) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.config.ConfigBuilder.buildConfigObject(ConfigBuilder.groovy:814) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.config.ConfigBuilder.build(ConfigBuilder.groovy:827) + at nextflow.cli.CmdRun.run(CmdRun.groovy:328) + at nextflow.cli.Launcher.run(Launcher.groovy:503) + at nextflow.cli.Launcher.main(Launcher.groovy:657) +Caused by: org.codehaus.groovy.control.MultipleCompilationErrorsException: startup failed: +/groovy/script/Script7BD980DA7F8768B9E9654BF7A071C3F7: 13: Unexpected input: '{' @ line 13, column 9. + process { + ^ + +1 error + + at org.codehaus.groovy.control.ErrorCollector.failIfErrors(ErrorCollector.java:292) + at org.codehaus.groovy.control.ErrorCollector.addFatalError(ErrorCollector.java:148) + at org.apache.groovy.parser.antlr4.AstBuilder.collectSyntaxError(AstBuilder.java:4753) + at org.apache.groovy.parser.antlr4.AstBuilder.access$100(AstBuilder.java:169) + at org.apache.groovy.parser.antlr4.AstBuilder$3.syntaxError(AstBuilder.java:4764) + at groovyjarjarantlr4.v4.runtime.ProxyErrorListener.syntaxError(ProxyErrorListener.java:44) + at groovyjarjarantlr4.v4.runtime.Parser.notifyErrorListeners(Parser.java:543) + at groovyjarjarantlr4.v4.runtime.DefaultErrorStrategy.notifyErrorListeners(DefaultErrorStrategy.java:154) + at org.apache.groovy.parser.antlr4.internal.DescriptiveErrorStrategy.reportInputMismatch(DescriptiveErrorStrategy.java:104) + at org.apache.groovy.parser.antlr4.internal.DescriptiveErrorStrategy.recover(DescriptiveErrorStrategy.java:55) + at org.apache.groovy.parser.antlr4.internal.DescriptiveErrorStrategy.recoverInline(DescriptiveErrorStrategy.java:68) + at groovyjarjarantlr4.v4.runtime.Parser.match(Parser.java:213) + at org.apache.groovy.parser.antlr4.GroovyParser.compilationUnit(GroovyParser.java:368) + at org.apache.groovy.parser.antlr4.AstBuilder.buildCST(AstBuilder.java:243) + at org.apache.groovy.parser.antlr4.AstBuilder.buildCST(AstBuilder.java:221) + at org.apache.groovy.parser.antlr4.AstBuilder.buildAST(AstBuilder.java:262) + at org.apache.groovy.parser.antlr4.Antlr4ParserPlugin.buildAST(Antlr4ParserPlugin.java:58) + at org.codehaus.groovy.control.SourceUnit.buildAST(SourceUnit.java:256) + at java.base/java.util.Iterator.forEachRemaining(Iterator.java:133) + at java.base/java.util.Spliterators$IteratorSpliterator.forEachRemaining(Spliterators.java:1845) + at java.base/java.util.stream.ReferencePipeline$Head.forEach(ReferencePipeline.java:762) + at org.codehaus.groovy.control.CompilationUnit.compile(CompilationUnit.java:663) + at ch.artecat.grengine.code.groovy.DefaultGroovyCompiler.compile(DefaultGroovyCompiler.java:229) + ... 63 common frames omitted diff --git a/modules/local/dmsanalysis/processgatk.nf b/modules/local/dmsanalysis/processgatk.nf new file mode 100644 index 0000000..f809024 --- /dev/null +++ b/modules/local/dmsanalysis/processgatk.nf @@ -0,0 +1,91 @@ +// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) +// https://github.com/nf-core/modules/tree/master/modules/nf-core/ +// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: +// https://nf-co.re/join +// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. +// All other parameters MUST be provided using the "task.ext" directive, see here: +// https://www.nextflow.io/docs/latest/process.html#ext +// where "task.ext" is a string. +// Any parameters that need to be evaluated in the context of a particular sample +// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. +// TODO nf-core: Software that can be piped together SHOULD be added to separate module files +// unless there is a run-time, storage advantage in implementing in this way +// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: +// bwa mem | samtools view -B -T ref.fasta +// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty +// list (`[]`) instead of a file can be used to work around this issue. + +process DMSANALYSIS_PROCESSGATK { + tag "$meta.id" + label 'process_single' + + // TODO nf-core: List required Conda package(s). + // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). + // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. + // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/YOUR-TOOL-HERE': + 'biocontainers/YOUR-TOOL-HERE' }" + + input: + // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" + // MUST be provided as an input via a Groovy Map called "meta". + // This information may not be required in some instances e.g. indexing reference genome files: + // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf + // TODO nf-core: Where applicable please provide/convert compressed files as input/output + // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. + tuple val(meta), path(bam) + + output: + // TODO nf-core: Named file extensions MUST be emitted for ALL output channels + tuple val(meta), path("*.bam"), emit: bam + // TODO nf-core: List additional required output channels/values here + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 + // If the software is unable to output a version number on the command-line then it can be manually specified + // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf + // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) + // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive + // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter + // using the Nextflow "task" variable e.g. "--threads $task.cpus" + // TODO nf-core: Please replace the example samtools command below with your module's command + // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) + """ + samtools \\ + sort \\ + $args \\ + -@ $task.cpus \\ + -o ${prefix}.bam \\ + -T $prefix \\ + $bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ + + stub: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: A stub section should mimic the execution of the original module as best as possible + // Have a look at the following examples: + // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 + // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 + """ + touch ${prefix}.bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ +} From 14396c4e00d00876d2e711460a619486b191ebbb Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 22:25:22 +0200 Subject: [PATCH 139/317] Update processgatk.nf --- modules/local/dmsanalysis/processgatk.nf | 96 +++++++----------------- 1 file changed, 29 insertions(+), 67 deletions(-) diff --git a/modules/local/dmsanalysis/processgatk.nf b/modules/local/dmsanalysis/processgatk.nf index f809024..d14fca4 100644 --- a/modules/local/dmsanalysis/processgatk.nf +++ b/modules/local/dmsanalysis/processgatk.nf @@ -1,91 +1,53 @@ -// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) -// https://github.com/nf-core/modules/tree/master/modules/nf-core/ -// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: -// https://nf-co.re/join -// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. -// All other parameters MUST be provided using the "task.ext" directive, see here: -// https://www.nextflow.io/docs/latest/process.html#ext -// where "task.ext" is a string. -// Any parameters that need to be evaluated in the context of a particular sample -// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. -// TODO nf-core: Software that can be piped together SHOULD be added to separate module files -// unless there is a run-time, storage advantage in implementing in this way -// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: -// bwa mem | samtools view -B -T ref.fasta -// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty -// list (`[]`) instead of a file can be used to work around this issue. - process DMSANALYSIS_PROCESSGATK { tag "$meta.id" label 'process_single' - // TODO nf-core: List required Conda package(s). - // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). - // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. - // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/YOUR-TOOL-HERE': - 'biocontainers/YOUR-TOOL-HERE' }" + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + + publishDir "${params.outdir}/intermediate_files", mode: 'copy' input: - // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" - // MUST be provided as an input via a Groovy Map called "meta". - // This information may not be required in some instances e.g. indexing reference genome files: - // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf - // TODO nf-core: Where applicable please provide/convert compressed files as input/output - // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. - tuple val(meta), path(bam) + tuple val(meta), path(variantCounts) + path possible_mutations + path aa_seq + val min_counts + path process_raw_gatk_script + path filter_by_library_script + path complete_gatk_script + path prepare_counts_heatmap_script output: - // TODO nf-core: Named file extensions MUST be emitted for ALL output channels - tuple val(meta), path("*.bam"), emit: bam - // TODO nf-core: List additional required output channels/values here - path "versions.yml" , emit: versions + tuple val(meta), + path("annotated_variantCounts.csv"), + path("variantCounts_filtered_by_library.csv"), + path("library_completed_variantCounts.csv"), + path("variantCounts_for_heatmaps.csv"), + emit: processed_variantCounts + + path "versions.yml", emit: versions when: task.ext.when == null || task.ext.when script: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 - // If the software is unable to output a version number on the command-line then it can be manually specified - // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf - // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) - // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive - // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter - // using the Nextflow "task" variable e.g. "--threads $task.cpus" - // TODO nf-core: Please replace the example samtools command below with your module's command - // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) """ - samtools \\ - sort \\ - $args \\ - -@ $task.cpus \\ - -o ${prefix}.bam \\ - -T $prefix \\ - $bam + Rscript -e "source('$process_raw_gatk_script'); process_raw_gatk('$variantCounts', 'annotated_variantCounts.csv')" + Rscript -e "source('$filter_by_library_script'); filter_gatk_by_codon_library('annotated_variantCounts.csv', '$possible_mutations', 'variantCounts_filtered_by_library.csv')" + Rscript -e "source('$complete_gatk_script'); complete_prefiltered_gatk('$possible_mutations', 'variantCounts_filtered_by_library.csv', 'library_completed_variantCounts.csv')" + Rscript -e "source('$prepare_counts_heatmap_script'); prepare_gatk_data_for_counts_heatmaps('variantCounts_filtered_by_library.csv', '$aa_seq', 'variantCounts_for_heatmaps.csv', $min_counts)" + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') cat <<-END_VERSIONS > versions.yml - "${task.process}": - dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + DMSANALYSIS_PROCESSGATK: + r-base: \$R_VERSION END_VERSIONS """ stub: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: A stub section should mimic the execution of the original module as best as possible - // Have a look at the following examples: - // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 - // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 """ - touch ${prefix}.bam - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') - END_VERSIONS + touch annotated_variantCounts.csv variantCounts_filtered_by_library.csv library_completed_variantCounts.csv variantCounts_for_heatmaps.csv + echo "DMSANALYSIS_PROCESSGATK:" > versions.yml + echo " stub-version: 0.0.0" >> versions.yml """ } From c748de59bedff9f2606b0a777db09763b70cf8ea Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 22:27:30 +0200 Subject: [PATCH 140/317] Update processgatk.nf --- modules/local/dmsanalysis/processgatk.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/processgatk.nf b/modules/local/dmsanalysis/processgatk.nf index d14fca4..6c60f1d 100644 --- a/modules/local/dmsanalysis/processgatk.nf +++ b/modules/local/dmsanalysis/processgatk.nf @@ -1,4 +1,4 @@ -process DMSANALYSIS_PROCESSGATK { +process DMSANALYSIS_PROCESS_GATK { tag "$meta.id" label 'process_single' From aec52167ceefd93b6eac39b7ae7b4be6374dd12b Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 22:27:36 +0200 Subject: [PATCH 141/317] Update dmscore.nf --- workflows/dmscore.nf | 11 +++++++++++ 1 file changed, 11 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 3de6fe1..bafbe2c 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -12,6 +12,7 @@ include { PREMERGE } from '../modules/local/bamprocessing/premerge include { GATK_SATURATIONMUTAGENESIS } from '../modules/local/gatk/saturationmutagenesis' include { DMSANALYSIS_AASEQ } from '../modules/local/dmsanalysis/aaseq' include { DMSANALYSIS_POSSIBLE_MUTATIONS } from '../modules/local/dmsanalysis/possiblemutations' +include { DMSANALYSIS_PROCESS_GATK } from '../modules/local/dmsanalysis/processgatk' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -194,6 +195,16 @@ workflow DMSCORE { ) ch_versions = ch_versions.mix(DMSANALYSIS_POSSIBLE_MUTATIONS.out.versions) + DMSANALYSIS_PROCESS_GATK( + GATK_SATURATIONMUTAGENESIS.out.variantCounts, + DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations, + DMSANALYSIS_AASEQ.out.aa_seq, + min_counts_ch, + process_raw_gatk_script_ch, + filter_by_library_script_ch, + complete_gatk_script_ch, + prepare_counts_heatmap_script_ch + ) emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html From 1c26e146e8c209b06af8a37863ad125321f2ccc6 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 22:28:44 +0200 Subject: [PATCH 142/317] Update modules.config --- conf/modules.config | 7 +++++++ 1 file changed, 7 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index a7da483..0dff51d 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -96,4 +96,11 @@ process { ] } + withName: 'DMSANALYSIS_PROCESS_GATK' { + publishDir = [ + path: "${params.outdir}/intermediate_files/processed_gatk_files", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } } From da2ed3561184cdcb2e7edc8ffae862faf1a9caa2 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 22:34:04 +0200 Subject: [PATCH 143/317] Update dmscore.nf --- workflows/dmscore.nf | 11 ++++++++--- 1 file changed, 8 insertions(+), 3 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index bafbe2c..d5e1ffa 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -179,6 +179,11 @@ workflow DMSCORE { min_counts_ch // min_counts (val) ) + gatk_variantCounts_ch = gatk_ch + .map { paths -> + paths.find { it.endsWith('gatk_output.variantCounts') } + } + DMSANALYSIS_AASEQ ( ch_fasta, reading_frame_ch, @@ -196,9 +201,9 @@ workflow DMSCORE { ch_versions = ch_versions.mix(DMSANALYSIS_POSSIBLE_MUTATIONS.out.versions) DMSANALYSIS_PROCESS_GATK( - GATK_SATURATIONMUTAGENESIS.out.variantCounts, - DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations, - DMSANALYSIS_AASEQ.out.aa_seq, + gatk_variantCounts_ch, + DMSANALYSIS_POSSIBLE_MUTATIONS.out, + DMSANALYSIS_AASEQ.out, min_counts_ch, process_raw_gatk_script_ch, filter_by_library_script_ch, From 588e669a1a4d98c464b1d89a944ddac514d7ff16 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 22:35:29 +0200 Subject: [PATCH 144/317] Update dmscore.nf --- workflows/dmscore.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index d5e1ffa..f46d559 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -202,8 +202,8 @@ workflow DMSCORE { DMSANALYSIS_PROCESS_GATK( gatk_variantCounts_ch, - DMSANALYSIS_POSSIBLE_MUTATIONS.out, - DMSANALYSIS_AASEQ.out, + DMSANALYSIS_POSSIBLE_MUTATIONS.out.csv, + DMSANALYSIS_AASEQ.out.txt, min_counts_ch, process_raw_gatk_script_ch, filter_by_library_script_ch, From 2ba8f36d1b5745bbb443181d0ad00e9dd0c8984f Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 22:37:19 +0200 Subject: [PATCH 145/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index f46d559..29be46d 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -179,7 +179,7 @@ workflow DMSCORE { min_counts_ch // min_counts (val) ) - gatk_variantCounts_ch = gatk_ch + gatk_variantCounts_ch = GATK_SATURATIONMUTAGENESIS.out .map { paths -> paths.find { it.endsWith('gatk_output.variantCounts') } } From 7d2361072bf69442786a44716ac6ec82865cac6b Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 22:54:56 +0200 Subject: [PATCH 146/317] Update dmscore.nf --- workflows/dmscore.nf | 12 ++++++------ 1 file changed, 6 insertions(+), 6 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 29be46d..15580ba 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -179,9 +179,9 @@ workflow DMSCORE { min_counts_ch // min_counts (val) ) - gatk_variantCounts_ch = GATK_SATURATIONMUTAGENESIS.out - .map { paths -> - paths.find { it.endsWith('gatk_output.variantCounts') } + gatk_variantCounts_ch = GATK_SATURATIONMUTAGENESIS.out.gatk_output + .map { meta, files -> + tuple(meta, files.find { it.name.endsWith("variantCounts") }) } DMSANALYSIS_AASEQ ( @@ -202,14 +202,14 @@ workflow DMSCORE { DMSANALYSIS_PROCESS_GATK( gatk_variantCounts_ch, - DMSANALYSIS_POSSIBLE_MUTATIONS.out.csv, - DMSANALYSIS_AASEQ.out.txt, + DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations, + DMSANALYSIS_AASEQ.out.aa_seq, min_counts_ch, process_raw_gatk_script_ch, filter_by_library_script_ch, complete_gatk_script_ch, prepare_counts_heatmap_script_ch - ) +) emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html From 9bc05d749705163c3afebf6386340a431fb8c796 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 23:01:39 +0200 Subject: [PATCH 147/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 15580ba..1f162bb 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -181,7 +181,7 @@ workflow DMSCORE { gatk_variantCounts_ch = GATK_SATURATIONMUTAGENESIS.out.gatk_output .map { meta, files -> - tuple(meta, files.find { it.name.endsWith("variantCounts") }) + tuple(meta, files.find { it.endsWith("variantCounts") }) } DMSANALYSIS_AASEQ ( From ca8af1c80c2fe0b3c4d8737582db48cf802cab87 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 23:07:58 +0200 Subject: [PATCH 148/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 1f162bb..95a1b27 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -209,7 +209,7 @@ workflow DMSCORE { filter_by_library_script_ch, complete_gatk_script_ch, prepare_counts_heatmap_script_ch -) + ) emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html From 10007ed0d1df320f6fc3e474522814566e5572a4 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 23:20:20 +0200 Subject: [PATCH 149/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index 3af1bf5..a126a83 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -12,6 +12,7 @@ process GATK_SATURATIONMUTAGENESIS { val min_counts output: + tuple val(meta), path("gatk_output.variantCounts"), emit: variantCounts tuple val(meta), path("gatk_output.*"), emit: gatk_output path "versions.yml", emit: versions From 90d2735f056dfa0ae230e74dafe76aa00eb883ea Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 23:21:10 +0200 Subject: [PATCH 150/317] Update dmscore.nf --- workflows/dmscore.nf | 7 +------ 1 file changed, 1 insertion(+), 6 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 95a1b27..bafbe2c 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -179,11 +179,6 @@ workflow DMSCORE { min_counts_ch // min_counts (val) ) - gatk_variantCounts_ch = GATK_SATURATIONMUTAGENESIS.out.gatk_output - .map { meta, files -> - tuple(meta, files.find { it.endsWith("variantCounts") }) - } - DMSANALYSIS_AASEQ ( ch_fasta, reading_frame_ch, @@ -201,7 +196,7 @@ workflow DMSCORE { ch_versions = ch_versions.mix(DMSANALYSIS_POSSIBLE_MUTATIONS.out.versions) DMSANALYSIS_PROCESS_GATK( - gatk_variantCounts_ch, + GATK_SATURATIONMUTAGENESIS.out.variantCounts, DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations, DMSANALYSIS_AASEQ.out.aa_seq, min_counts_ch, From 0a6c3912421f21a21ee6f1d8a0cd3abe4e755149 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 23:38:12 +0200 Subject: [PATCH 151/317] Update dmscore.nf --- workflows/dmscore.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index bafbe2c..348d133 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -197,8 +197,8 @@ workflow DMSCORE { DMSANALYSIS_PROCESS_GATK( GATK_SATURATIONMUTAGENESIS.out.variantCounts, - DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations, - DMSANALYSIS_AASEQ.out.aa_seq, + DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations.map{ it[1] }, + DMSANALYSIS_AASEQ.out.aa_seq.map{ it[1] }, min_counts_ch, process_raw_gatk_script_ch, filter_by_library_script_ch, From e34671ac44aee393950ca2287f499f474f150a59 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 23:41:09 +0200 Subject: [PATCH 152/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 1 - 1 file changed, 1 deletion(-) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index a126a83..3af1bf5 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -12,7 +12,6 @@ process GATK_SATURATIONMUTAGENESIS { val min_counts output: - tuple val(meta), path("gatk_output.variantCounts"), emit: variantCounts tuple val(meta), path("gatk_output.*"), emit: gatk_output path "versions.yml", emit: versions From f9c4d0cb6c834370ce6f410ccf4bd642f8a3f42a Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 7 May 2025 23:42:36 +0200 Subject: [PATCH 153/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index 3af1bf5..a126a83 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -12,6 +12,7 @@ process GATK_SATURATIONMUTAGENESIS { val min_counts output: + tuple val(meta), path("gatk_output.variantCounts"), emit: variantCounts tuple val(meta), path("gatk_output.*"), emit: gatk_output path "versions.yml", emit: versions From 5489d957dc25b6426f6448adbd9847f4c656ed26 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Thu, 8 May 2025 00:07:43 +0200 Subject: [PATCH 154/317] Update saturationmutagenesis.nf --- modules/local/gatk/saturationmutagenesis.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/gatk/saturationmutagenesis.nf b/modules/local/gatk/saturationmutagenesis.nf index a126a83..2616347 100644 --- a/modules/local/gatk/saturationmutagenesis.nf +++ b/modules/local/gatk/saturationmutagenesis.nf @@ -12,7 +12,7 @@ process GATK_SATURATIONMUTAGENESIS { val min_counts output: - tuple val(meta), path("gatk_output.variantCounts"), emit: variantCounts + tuple val(meta), path("gatk_output.variantCounts"), emit: variantCounts // to access the output tuple val(meta), path("gatk_output.*"), emit: gatk_output path "versions.yml", emit: versions From 1c5bb4f2090cf5004aa82ae1f897e95ff0b33ef9 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Thu, 8 May 2025 21:21:22 +0200 Subject: [PATCH 155/317] Update dmscore.nf --- workflows/dmscore.nf | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 348d133..3c1bccf 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -206,6 +206,11 @@ workflow DMSCORE { prepare_counts_heatmap_script_ch ) + annotated_variantCounts_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, a) } + variantCounts_filtered_by_library_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, b) } + library_completed_variantCounts_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, c) } + variantCounts_for_heatmaps_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, d) } + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From a8539f542eb8093d331834a5a0823ed6be43ad96 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Thu, 8 May 2025 21:33:46 +0200 Subject: [PATCH 156/317] Update visualization.nf --- modules/local/visualization/visualization.nf | 83 ++++---------------- 1 file changed, 15 insertions(+), 68 deletions(-) diff --git a/modules/local/visualization/visualization.nf b/modules/local/visualization/visualization.nf index 7ddcc83..07e130d 100644 --- a/modules/local/visualization/visualization.nf +++ b/modules/local/visualization/visualization.nf @@ -1,91 +1,38 @@ -// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) -// https://github.com/nf-core/modules/tree/master/modules/nf-core/ -// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: -// https://nf-co.re/join -// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. -// All other parameters MUST be provided using the "task.ext" directive, see here: -// https://www.nextflow.io/docs/latest/process.html#ext -// where "task.ext" is a string. -// Any parameters that need to be evaluated in the context of a particular sample -// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. -// TODO nf-core: Software that can be piped together SHOULD be added to separate module files -// unless there is a run-time, storage advantage in implementing in this way -// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: -// bwa mem | samtools view -B -T ref.fasta -// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty -// list (`[]`) instead of a file can be used to work around this issue. - -process DMSANALYSIS_VISUALIZATION { +process DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV { tag "$meta.id" label 'process_single' - // TODO nf-core: List required Conda package(s). - // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). - // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. - // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/YOUR-TOOL-HERE': - 'biocontainers/YOUR-TOOL-HERE' }" + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" input: - // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" - // MUST be provided as an input via a Groovy Map called "meta". - // This information may not be required in some instances e.g. indexing reference genome files: - // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf - // TODO nf-core: Where applicable please provide/convert compressed files as input/output - // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. - tuple val(meta), path(bam) + tuple val(meta), path(variantCounts_for_heatmaps) + val min_counts + path script // counts_per_cov_heatmap.R output: - // TODO nf-core: Named file extensions MUST be emitted for ALL output channels - tuple val(meta), path("*.bam"), emit: bam - // TODO nf-core: List additional required output channels/values here - path "versions.yml" , emit: versions + tuple val(meta), path("counts_per_cov_heatmap.pdf"), emit: counts_per_cov_heatmap + path "versions.yml", emit: versions when: task.ext.when == null || task.ext.when script: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 - // If the software is unable to output a version number on the command-line then it can be manually specified - // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf - // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) - // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive - // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter - // using the Nextflow "task" variable e.g. "--threads $task.cpus" - // TODO nf-core: Please replace the example samtools command below with your module's command - // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) """ - samtools \\ - sort \\ - $args \\ - -@ $task.cpus \\ - -o ${prefix}.bam \\ - -T $prefix \\ - $bam + Rscript -e "source('$script'); counts_per_cov_heatmap('$variantCounts_for_heatmaps', $min_counts, 'counts_per_cov_heatmap.pdf')" + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') cat <<-END_VERSIONS > versions.yml - "${task.process}": - dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') + DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV: + r-base: \$R_VERSION END_VERSIONS """ stub: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: A stub section should mimic the execution of the original module as best as possible - // Have a look at the following examples: - // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 - // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 """ - touch ${prefix}.bam - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') - END_VERSIONS + touch counts_per_cov_heatmap.pdf + echo "DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV:" > versions.yml + echo " stub-version: 0.0.0" >> versions.yml """ } + From 658fca27152b4874aa5dacf0240a2e6cb1a89764 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Thu, 8 May 2025 21:35:43 +0200 Subject: [PATCH 157/317] Update dmscore.nf --- workflows/dmscore.nf | 6 ++++++ 1 file changed, 6 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 3c1bccf..980a496 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -211,6 +211,12 @@ workflow DMSCORE { library_completed_variantCounts_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, c) } variantCounts_for_heatmaps_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, d) } + DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV( + variantCounts_for_heatmaps_ch, + min_counts_ch, + counts_per_cov_heatmap_script_ch + ) + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From 7f981ad2e9143db9afa7711e9d7041fd9905b95a Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Thu, 8 May 2025 21:38:08 +0200 Subject: [PATCH 158/317] Update dmscore.nf --- workflows/dmscore.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 980a496..56c1d99 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -13,6 +13,7 @@ include { GATK_SATURATIONMUTAGENESIS } from '../modules/local/gatk/satu include { DMSANALYSIS_AASEQ } from '../modules/local/dmsanalysis/aaseq' include { DMSANALYSIS_POSSIBLE_MUTATIONS } from '../modules/local/dmsanalysis/possiblemutations' include { DMSANALYSIS_PROCESS_GATK } from '../modules/local/dmsanalysis/processgatk' +include { DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV } from '../modules/local/dmsanalysis/visualization/visualization' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' From 4f9f7b91c820d8f5a59c91028f1a819a33bcb14c Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Thu, 8 May 2025 21:40:03 +0200 Subject: [PATCH 159/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 56c1d99..4b07172 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -13,7 +13,7 @@ include { GATK_SATURATIONMUTAGENESIS } from '../modules/local/gatk/satu include { DMSANALYSIS_AASEQ } from '../modules/local/dmsanalysis/aaseq' include { DMSANALYSIS_POSSIBLE_MUTATIONS } from '../modules/local/dmsanalysis/possiblemutations' include { DMSANALYSIS_PROCESS_GATK } from '../modules/local/dmsanalysis/processgatk' -include { DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV } from '../modules/local/dmsanalysis/visualization/visualization' +include { DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV } from '../modules/local/visualization/visualization' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' From 420319aaf0950094ebf1afabf9c6c0b2f54c9115 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Thu, 8 May 2025 21:46:21 +0200 Subject: [PATCH 160/317] Update visualization.nf --- modules/local/visualization/visualization.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/modules/local/visualization/visualization.nf b/modules/local/visualization/visualization.nf index 07e130d..f93e868 100644 --- a/modules/local/visualization/visualization.nf +++ b/modules/local/visualization/visualization.nf @@ -1,4 +1,4 @@ -process DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV { +process VISUALIZATION_COUNTS_PER_COV { tag "$meta.id" label 'process_single' @@ -23,7 +23,7 @@ process DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV { R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') cat <<-END_VERSIONS > versions.yml - DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV: + VISUALIZATION_COUNTS_PER_COV: r-base: \$R_VERSION END_VERSIONS """ @@ -31,7 +31,7 @@ process DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV { stub: """ touch counts_per_cov_heatmap.pdf - echo "DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV:" > versions.yml + echo "VISUALIZATION_COUNTS_PER_COV:" > versions.yml echo " stub-version: 0.0.0" >> versions.yml """ } From 0a084c71f7bafb1d2dc1de5f669b16584c4fe74a Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Thu, 8 May 2025 21:50:51 +0200 Subject: [PATCH 161/317] Update modules.config --- conf/modules.config | 9 ++++----- 1 file changed, 4 insertions(+), 5 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 0dff51d..c613dcc 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -88,17 +88,16 @@ process { ] } - withName: 'DMSANALYSIS_AASEQ' { + withName: 'DMSANALYSIS_PROCESS_GATK' { publishDir = [ - path: "${params.outdir}/intermediate_files", + path: "${params.outdir}/intermediate_files/processed_gatk_files", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } - - withName: 'DMSANALYSIS_PROCESS_GATK' { + withName: 'VISUALIZATION_COUNTS_PER_COV' { publishDir = [ - path: "${params.outdir}/intermediate_files/processed_gatk_files", + path: "${params.outdir}/final_plots", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] From d49f69421eebf8856aa02c67c89562b8e8b617c0 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Thu, 8 May 2025 21:53:43 +0200 Subject: [PATCH 162/317] Update dmscore.nf --- workflows/dmscore.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 4b07172..5a23114 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -13,7 +13,7 @@ include { GATK_SATURATIONMUTAGENESIS } from '../modules/local/gatk/satu include { DMSANALYSIS_AASEQ } from '../modules/local/dmsanalysis/aaseq' include { DMSANALYSIS_POSSIBLE_MUTATIONS } from '../modules/local/dmsanalysis/possiblemutations' include { DMSANALYSIS_PROCESS_GATK } from '../modules/local/dmsanalysis/processgatk' -include { DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV } from '../modules/local/visualization/visualization' +include { VISUALIZATION_COUNTS_PER_COV } from '../modules/local/visualization/visualization' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -212,7 +212,7 @@ workflow DMSCORE { library_completed_variantCounts_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, c) } variantCounts_for_heatmaps_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, d) } - DMSANALYSIS_VISUALIZATION_COUNTS_PER_COV( + VISUALIZATION_COUNTS_PER_COV( variantCounts_for_heatmaps_ch, min_counts_ch, counts_per_cov_heatmap_script_ch From 3d3a8c932b81d1be5bc9ab8fdee23b482c0f6256 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 10:38:06 +0200 Subject: [PATCH 163/317] Update visualization.nf --- modules/local/visualization/visualization.nf | 37 ++++++++++++++++++++ 1 file changed, 37 insertions(+) diff --git a/modules/local/visualization/visualization.nf b/modules/local/visualization/visualization.nf index f93e868..845c639 100644 --- a/modules/local/visualization/visualization.nf +++ b/modules/local/visualization/visualization.nf @@ -36,3 +36,40 @@ process VISUALIZATION_COUNTS_PER_COV { """ } +process VISUALIZATION_COUNTS_HEATMAP { + tag "$meta.id" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + + input: + tuple val(meta), path(variantCounts_for_heatmaps) + val min_counts + path script // counts_heatmap.R + + output: + tuple val(meta), path("counts_heatmap.pdf"), emit: counts_heatmap + path "versions.yml", emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + """ + Rscript -e "source('$script'); counts_heatmap('$variantCounts_for_heatmaps', $min_counts, 'counts_heatmap.pdf')" + + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + cat <<-END_VERSIONS > versions.yml + VISUALIZATION_COUNTS_PER_COV: + r-base: \$R_VERSION + END_VERSIONS + """ + + stub: + """ + touch counts_heatmap.pdf + echo "VISUALIZATION_COUNTS_HEATMAP:" > versions.yml + echo " stub-version: 0.0.0" >> versions.yml + """ +} From 3f7e6e1c134ea732d9062f62cc602c998f118f76 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 10:39:52 +0200 Subject: [PATCH 164/317] Update modules.config --- conf/modules.config | 9 +++++++++ 1 file changed, 9 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index c613dcc..fbd16f9 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -95,6 +95,7 @@ process { saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } + withName: 'VISUALIZATION_COUNTS_PER_COV' { publishDir = [ path: "${params.outdir}/final_plots", @@ -102,4 +103,12 @@ process { saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } + + withName: 'VISUALIZATION_COUNTS_HEATMAP' { + publishDir = [ + path: "${params.outdir}/final_plots", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } } From 9851e09098d3ce4b1ac9a3f25c9ab0cceff08c12 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 10:39:56 +0200 Subject: [PATCH 165/317] Update dmscore.nf --- workflows/dmscore.nf | 7 +++++++ 1 file changed, 7 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 5a23114..7e058ae 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -14,6 +14,7 @@ include { DMSANALYSIS_AASEQ } from '../modules/local/dmsanalysis/aaseq' include { DMSANALYSIS_POSSIBLE_MUTATIONS } from '../modules/local/dmsanalysis/possiblemutations' include { DMSANALYSIS_PROCESS_GATK } from '../modules/local/dmsanalysis/processgatk' include { VISUALIZATION_COUNTS_PER_COV } from '../modules/local/visualization/visualization' +include { VISUALIZATION_COUNTS_HEATMAP } from '../modules/local/visualization/visualization' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -218,6 +219,12 @@ workflow DMSCORE { counts_per_cov_heatmap_script_ch ) + VISUALIZATION_COUNTS_HEATMAP( + variantCounts_for_heatmaps_ch, + min_counts_ch, + counts_per_cov_heatmap_script_ch + ) + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From 8fd57eb43bf729d2e65b7e738552e2597fe58db5 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 10:43:39 +0200 Subject: [PATCH 166/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 7e058ae..7a04756 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -222,7 +222,7 @@ workflow DMSCORE { VISUALIZATION_COUNTS_HEATMAP( variantCounts_for_heatmaps_ch, min_counts_ch, - counts_per_cov_heatmap_script_ch + counts_heatmap_script_ch ) emit: From 0ae331ea2bb1ecd5d7a0bc8604bbf39f8f15a4ec Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 10:50:58 +0200 Subject: [PATCH 167/317] Update visualization.nf --- modules/local/visualization/visualization.nf | 41 ++++++++++++++++++++ 1 file changed, 41 insertions(+) diff --git a/modules/local/visualization/visualization.nf b/modules/local/visualization/visualization.nf index 845c639..0a6be43 100644 --- a/modules/local/visualization/visualization.nf +++ b/modules/local/visualization/visualization.nf @@ -73,3 +73,44 @@ process VISUALIZATION_COUNTS_HEATMAP { echo " stub-version: 0.0.0" >> versions.yml """ } + +process VISUALIZATION_GLOBAL_POS_BIASES_COUNTS { + tag "$meta.id" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + + input: + tuple val(meta), path(variantCounts_filtered_by_library) + path aa_seq + val sliding_window_size + path script // global_position_biases_counts_and_counts_per_cov.R + + output: + tuple val(meta), path("rolling_counts.pdf"), emit: rolling_counts + tuple val(meta), path("rolling_counts_per_cov.pdf"), emit: rolling_counts_per_cov + path "versions.yml", emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + """ + Rscript -e "source('$script'); position_biases('$variantCounts_filtered_by_library', '$aa_seq', $sliding_window_size)" + + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + cat <<-END_VERSIONS > versions.yml + VISUALIZATION_COUNTS_PER_COV: + r-base: \$R_VERSION + END_VERSIONS + """ + + stub: + """ + touch rolling_counts.pdf + touch rolling_counts_per_cov.pdf + echo "VISUALIZATION_COUNTS_HEATMAP:" > versions.yml + echo " stub-version: 0.0.0" >> versions.yml + """ +} From 7d9f84a874c3663a1b7fe7d634d81291f4b10c7e Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 10:56:31 +0200 Subject: [PATCH 168/317] Update dmscore.nf --- workflows/dmscore.nf | 15 +++++++++++++++ 1 file changed, 15 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 7a04756..860f406 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -15,6 +15,7 @@ include { DMSANALYSIS_POSSIBLE_MUTATIONS } from '../modules/local/dmsanalys include { DMSANALYSIS_PROCESS_GATK } from '../modules/local/dmsanalysis/processgatk' include { VISUALIZATION_COUNTS_PER_COV } from '../modules/local/visualization/visualization' include { VISUALIZATION_COUNTS_HEATMAP } from '../modules/local/visualization/visualization' +include { VISUALIZATION_GLOBAL_POS_BIASES_COUNTS } from '../modules/local/visualization/visualization' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -30,6 +31,7 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library +params.sliding_window_size = params.sliding_window_size ?: 10 // sliding window size to flatten graphs in plots (e.g. GLOBAL_POS_BIASES_COUNTS function) // Define fasta file as channel (e.g. for BWA index) Channel @@ -57,6 +59,12 @@ Channel .value(params.mutagenesis_type) .set { mutagenesis_type_ch } +// Define sliding_window_size as channel (e.g. for GLOBAL_POS_BIASES_COUNTS function) -> if not set - default: 10 +Channel + .value(params.sliding_window_size) + .set { sliding_window_size_ch } + + // Define R scripts as channels Channel.fromPath("modules/local/dmsanalysis/bin/SeqDepth_simulation.R", checkIfExists: true).set { seqdepth_simulation_script_ch } Channel.fromPath("modules/local/dmsanalysis/bin/aa_seq.R", checkIfExists: true).set { aa_seq_script_ch } @@ -225,6 +233,13 @@ workflow DMSCORE { counts_heatmap_script_ch ) + VISUALIZATION_GLOBAL_POS_BIASES_COUNTS( + variantCounts_filtered_by_library_ch, + DMSANALYSIS_AASEQ.out.aa_seq.map{ it[1] }, + sliding_window_size_ch, + global_bias_counts_cov_script_ch + ) + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From 6835f6abd29075d0b96c0295029378dff0a4ddd9 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 10:57:24 +0200 Subject: [PATCH 169/317] Update modules.config --- conf/modules.config | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index fbd16f9..59aab27 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -111,4 +111,12 @@ process { saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } + + withName: 'VISUALIZATION_GLOBAL_POS_BIASES_COUNTS' { + publishDir = [ + path: "${params.outdir}/final_plots", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } } From c7d93f2b0773d0b09473301c3ab51013b464ee41 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:01:49 +0200 Subject: [PATCH 170/317] Update visualization.nf --- modules/local/visualization/visualization.nf | 40 ++++++++++++++++++++ 1 file changed, 40 insertions(+) diff --git a/modules/local/visualization/visualization.nf b/modules/local/visualization/visualization.nf index 0a6be43..0cc720c 100644 --- a/modules/local/visualization/visualization.nf +++ b/modules/local/visualization/visualization.nf @@ -114,3 +114,43 @@ process VISUALIZATION_GLOBAL_POS_BIASES_COUNTS { echo " stub-version: 0.0.0" >> versions.yml """ } + +process VISUALIZATION_GLOBAL_POS_BIASES_COV { + tag "$meta.id" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + + input: + tuple val(meta), path(variantCounts_filtered_by_library) + path aa_seq + val sliding_window_size + val aimed_cov + path script // global_position_biases_counts_and_counts_per_cov.R + + output: + tuple val(meta), path("rolling_coverage.pdf"), emit: rolling_coverage + path "versions.yml", emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + """ + Rscript -e "source('$script'); position_biases('$variantCounts_filtered_by_library', '$aa_seq', $sliding_window_size, 'rolling_coverage.pdf', $aimed_cov)" + + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + cat <<-END_VERSIONS > versions.yml + VISUALIZATION_COUNTS_PER_COV: + r-base: \$R_VERSION + END_VERSIONS + """ + + stub: + """ + touch rolling_coverage.pdf + echo "VISUALIZATION_COUNTS_HEATMAP:" > versions.yml + echo " stub-version: 0.0.0" >> versions.yml + """ +} From bc962ff60fc79acd5753fe03233a83996b2fbae1 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:02:13 +0200 Subject: [PATCH 171/317] Update modules.config --- conf/modules.config | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index 59aab27..3267bd2 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -119,4 +119,12 @@ process { saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } + + withName: 'VISUALIZATION_GLOBAL_POS_BIASES_COVERAGE' { + publishDir = [ + path: "${params.outdir}/final_plots", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } } From 286f0dbf2eaf3fa84b2444b835abbb16d5931402 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:06:37 +0200 Subject: [PATCH 172/317] Update dmscore.nf --- workflows/dmscore.nf | 15 +++++++++++++++ 1 file changed, 15 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 860f406..4f9b66c 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -16,6 +16,7 @@ include { DMSANALYSIS_PROCESS_GATK } from '../modules/local/dmsanalysis/pro include { VISUALIZATION_COUNTS_PER_COV } from '../modules/local/visualization/visualization' include { VISUALIZATION_COUNTS_HEATMAP } from '../modules/local/visualization/visualization' include { VISUALIZATION_GLOBAL_POS_BIASES_COUNTS } from '../modules/local/visualization/visualization' +include { VISUALIZATION_GLOBAL_POS_BIASES_COVERAGE } from '../modules/local/visualization/visualization' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -32,6 +33,7 @@ params.min_counts = params.min_counts ?: 3 params.mutagenesis_type = params.mutagenesis_type ?: 'nnk' // default library is set to nnk params.custom_codon_library = params.custom_codon_library ?: '/NULL' // when mutagenesis_type is set to >>custom<< this variable has to be path to .txt with custom library params.sliding_window_size = params.sliding_window_size ?: 10 // sliding window size to flatten graphs in plots (e.g. GLOBAL_POS_BIASES_COUNTS function) +params.aimed_cov = params.aimed_cov ?: 100 // aimed coverage (assuming equal spread) to visualize threshold in plots // Define fasta file as channel (e.g. for BWA index) Channel @@ -64,6 +66,11 @@ Channel .value(params.sliding_window_size) .set { sliding_window_size_ch } +// Define aimed_cov as channel (e.g. for GLOBAL_POS_BIASES_COVERAGE function) -> if not set - default: 100 +Channel + .value(params.aimed_cov) + .set { aimed_cov_ch } + // Define R scripts as channels Channel.fromPath("modules/local/dmsanalysis/bin/SeqDepth_simulation.R", checkIfExists: true).set { seqdepth_simulation_script_ch } @@ -240,6 +247,14 @@ workflow DMSCORE { global_bias_counts_cov_script_ch ) + VISUALIZATION_GLOBAL_POS_BIASES_COVERAGE( + variantCounts_filtered_by_library_ch, + DMSANALYSIS_AASEQ.out.aa_seq.map{ it[1] }, + sliding_window_size_ch, + aimed_cov_ch, + global_bias_cov_script_ch + ) + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From 1cf11a73405607c5f192cc187fa844cc6ebd7d8c Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:08:56 +0200 Subject: [PATCH 173/317] Update dmscore.nf --- workflows/dmscore.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 4f9b66c..cee024e 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -16,7 +16,7 @@ include { DMSANALYSIS_PROCESS_GATK } from '../modules/local/dmsanalysis/pro include { VISUALIZATION_COUNTS_PER_COV } from '../modules/local/visualization/visualization' include { VISUALIZATION_COUNTS_HEATMAP } from '../modules/local/visualization/visualization' include { VISUALIZATION_GLOBAL_POS_BIASES_COUNTS } from '../modules/local/visualization/visualization' -include { VISUALIZATION_GLOBAL_POS_BIASES_COVERAGE } from '../modules/local/visualization/visualization' +include { VISUALIZATION_GLOBAL_POS_BIASES_COV } from '../modules/local/visualization/visualization' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -247,7 +247,7 @@ workflow DMSCORE { global_bias_counts_cov_script_ch ) - VISUALIZATION_GLOBAL_POS_BIASES_COVERAGE( + VISUALIZATION_GLOBAL_POS_BIASES_COV( variantCounts_filtered_by_library_ch, DMSANALYSIS_AASEQ.out.aa_seq.map{ it[1] }, sliding_window_size_ch, From bce67509238f59de66a8e343a3742e57f7fe7d7e Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:09:01 +0200 Subject: [PATCH 174/317] Update modules.config --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index 3267bd2..5a9068c 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -120,7 +120,7 @@ process { ] } - withName: 'VISUALIZATION_GLOBAL_POS_BIASES_COVERAGE' { + withName: 'VISUALIZATION_GLOBAL_POS_BIASES_COV' { publishDir = [ path: "${params.outdir}/final_plots", mode: 'copy', From 670204c8dfa18784801acefb338fae02cfe26b95 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:16:18 +0200 Subject: [PATCH 175/317] Update visualization.nf --- modules/local/visualization/visualization.nf | 39 ++++++++++++++++++++ 1 file changed, 39 insertions(+) diff --git a/modules/local/visualization/visualization.nf b/modules/local/visualization/visualization.nf index 0cc720c..5f059bf 100644 --- a/modules/local/visualization/visualization.nf +++ b/modules/local/visualization/visualization.nf @@ -154,3 +154,42 @@ process VISUALIZATION_GLOBAL_POS_BIASES_COV { echo " stub-version: 0.0.0" >> versions.yml """ } + +process VISUALIZATION_LOGDIFF { + tag "$meta.id" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + + input: + tuple val(meta), path(library_completed_variantCounts) + path script // global_position_biases_counts_and_counts_per_cov.R + + output: + tuple val(meta), path("logdiff_plot.pdf"), emit: logdiff_plot + tuple val(meta), path("logdiff_varying_bases.pdf"), emit: logdiff_varying_bases + path "versions.yml", emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + """ + Rscript -e "source('$script'); logdiff_plot('$library_completed_variantCounts')" + + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + cat <<-END_VERSIONS > versions.yml + VISUALIZATION_COUNTS_PER_COV: + r-base: \$R_VERSION + END_VERSIONS + """ + + stub: + """ + touch logdiff_plot.pdf + touch logdiff_varying_bases.pdf + echo "VISUALIZATION_COUNTS_HEATMAP:" > versions.yml + echo " stub-version: 0.0.0" >> versions.yml + """ +} From 4c9957ddde951c5f25f5b80bccc318f74bd39cd8 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:17:24 +0200 Subject: [PATCH 176/317] Update dmscore.nf --- workflows/dmscore.nf | 6 ++++++ 1 file changed, 6 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index cee024e..43f8231 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -17,6 +17,7 @@ include { VISUALIZATION_COUNTS_PER_COV } from '../modules/local/visualizati include { VISUALIZATION_COUNTS_HEATMAP } from '../modules/local/visualization/visualization' include { VISUALIZATION_GLOBAL_POS_BIASES_COUNTS } from '../modules/local/visualization/visualization' include { VISUALIZATION_GLOBAL_POS_BIASES_COV } from '../modules/local/visualization/visualization' +include { VISUALIZATION_LOGDIFF } from '../modules/local/visualization/visualization' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -255,6 +256,11 @@ workflow DMSCORE { global_bias_cov_script_ch ) + VISUALIZATION_LOGDIFF( + library_completed_variantCounts_ch, + logdiff_script_ch + ) + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From 182728b140f2d5b7d710afeec806eb7da2502db8 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:17:48 +0200 Subject: [PATCH 177/317] Update modules.config --- conf/modules.config | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index 5a9068c..9db1d9a 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -127,4 +127,12 @@ process { saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } + + withName: 'VISUALIZATION_LOGDIFF' { + publishDir = [ + path: "${params.outdir}/final_plots", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } } From d4fcba350c82e0e695abe25e903d6a28e2ecdfb1 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:24:19 +0200 Subject: [PATCH 178/317] Update visualization.nf --- modules/local/visualization/visualization.nf | 43 +++++++++++++++++++- 1 file changed, 41 insertions(+), 2 deletions(-) diff --git a/modules/local/visualization/visualization.nf b/modules/local/visualization/visualization.nf index 5f059bf..d7e9087 100644 --- a/modules/local/visualization/visualization.nf +++ b/modules/local/visualization/visualization.nf @@ -127,7 +127,7 @@ process VISUALIZATION_GLOBAL_POS_BIASES_COV { path aa_seq val sliding_window_size val aimed_cov - path script // global_position_biases_counts_and_counts_per_cov.R + path script // R script output: tuple val(meta), path("rolling_coverage.pdf"), emit: rolling_coverage @@ -164,7 +164,7 @@ process VISUALIZATION_LOGDIFF { input: tuple val(meta), path(library_completed_variantCounts) - path script // global_position_biases_counts_and_counts_per_cov.R + path script // R script output: tuple val(meta), path("logdiff_plot.pdf"), emit: logdiff_plot @@ -193,3 +193,42 @@ process VISUALIZATION_LOGDIFF { echo " stub-version: 0.0.0" >> versions.yml """ } + +process VISUALIZATION_SEQDEPTH { + tag "$meta.id" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + + input: + tuple val(meta), path(variantCounts_filtered_by_library) + path possible_mutations + val min_counts + path script // R script + + output: + tuple val(meta), path("SeqDepth.pdf"), emit: SeqDepth + path "versions.yml", emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + """ + Rscript -e "source('$script'); SeqDepth_simulation_plot('$variantCounts_filtered_by_library', '$possible_mutations', 'SeqDepth.pdf', 0.01, $min_counts)" + + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + cat <<-END_VERSIONS > versions.yml + VISUALIZATION_COUNTS_PER_COV: + r-base: \$R_VERSION + END_VERSIONS + """ + + stub: + """ + touch SeqDepth.pdf + echo "VISUALIZATION_COUNTS_HEATMAP:" > versions.yml + echo " stub-version: 0.0.0" >> versions.yml + """ +} From 06fa544c3567420831ca383f209668d219dd4a02 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:26:07 +0200 Subject: [PATCH 179/317] Update dmscore.nf --- workflows/dmscore.nf | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 43f8231..d1937f6 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -18,6 +18,7 @@ include { VISUALIZATION_COUNTS_HEATMAP } from '../modules/local/visualizati include { VISUALIZATION_GLOBAL_POS_BIASES_COUNTS } from '../modules/local/visualization/visualization' include { VISUALIZATION_GLOBAL_POS_BIASES_COV } from '../modules/local/visualization/visualization' include { VISUALIZATION_LOGDIFF } from '../modules/local/visualization/visualization' +include { VISUALIZATION_SEQDEPTH } from '../modules/local/visualization/visualization' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -261,6 +262,13 @@ workflow DMSCORE { logdiff_script_ch ) + VISUALIZATION_SEQDEPTH( + variantCounts_filtered_by_library_ch, + DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations.map{ it[1] }, + min_counts_ch, + seqdepth_simulation_script_ch + ) + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From b68948dba681e54c091c1492ef5dbab79a27844f Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:26:36 +0200 Subject: [PATCH 180/317] Update modules.config --- conf/modules.config | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index 9db1d9a..eb49bec 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -135,4 +135,12 @@ process { saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } + + withName: 'VISUALIZATION_SEQDEPTH' { + publishDir = [ + path: "${params.outdir}/final_plots", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } } From 672da83dfcab2a83ebe0658b8c034a64044a29c4 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:28:34 +0200 Subject: [PATCH 181/317] Update visualization.nf --- modules/local/visualization/visualization.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/visualization/visualization.nf b/modules/local/visualization/visualization.nf index d7e9087..5bf6cd2 100644 --- a/modules/local/visualization/visualization.nf +++ b/modules/local/visualization/visualization.nf @@ -196,7 +196,7 @@ process VISUALIZATION_LOGDIFF { process VISUALIZATION_SEQDEPTH { tag "$meta.id" - label 'process_single' + label 'process_high' conda "${moduleDir}/environment.yml" container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" From 44f8ae2f5f7bfca60989350b847502db51138f06 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:33:58 +0200 Subject: [PATCH 182/317] Update dmscore.nf --- workflows/dmscore.nf | 17 +++++++++++++---- 1 file changed, 13 insertions(+), 4 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index d1937f6..b0a71cd 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -36,6 +36,8 @@ params.mutagenesis_type = params.mutagenesis_type ?: 'nnk' params.custom_codon_library = params.custom_codon_library ?: '/NULL' // when mutagenesis_type is set to >>custom<< this variable has to be path to .txt with custom library params.sliding_window_size = params.sliding_window_size ?: 10 // sliding window size to flatten graphs in plots (e.g. GLOBAL_POS_BIASES_COUNTS function) params.aimed_cov = params.aimed_cov ?: 100 // aimed coverage (assuming equal spread) to visualize threshold in plots +params.run_seqdepth = params.run_seqdepth ?: false // creating seqdepth simulation plot, is computationally quite heavy. per default disabled. + // Define fasta file as channel (e.g. for BWA index) Channel @@ -73,6 +75,11 @@ Channel .value(params.aimed_cov) .set { aimed_cov_ch } +// Define if seqdepth plot should be executed +Channel + .value(params.run_seqdepth) + .set { run_seqdepth_ch } + // Define R scripts as channels Channel.fromPath("modules/local/dmsanalysis/bin/SeqDepth_simulation.R", checkIfExists: true).set { seqdepth_simulation_script_ch } @@ -262,12 +269,14 @@ workflow DMSCORE { logdiff_script_ch ) + if (params.run_seqdepth) { VISUALIZATION_SEQDEPTH( - variantCounts_filtered_by_library_ch, - DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations.map{ it[1] }, - min_counts_ch, - seqdepth_simulation_script_ch + variantCounts_filtered_by_library_ch, + DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations.map{ it[1] }, + min_counts_ch, + seqdepth_simulation_script_ch ) + } emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html From 9895f02dbf3adb3c1d1484268e97258cd5cfe92b Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:35:44 +0200 Subject: [PATCH 183/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index b0a71cd..801152b 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -275,7 +275,7 @@ workflow DMSCORE { DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations.map{ it[1] }, min_counts_ch, seqdepth_simulation_script_ch - ) + ) } emit: From 600635ee737cd79e39b7273a24f1a623e855f037 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Fri, 9 May 2025 11:43:11 +0200 Subject: [PATCH 184/317] gatk to dimsum --- log.txt | 697 ++++++++++++++++++++++++----- modules/local/gatk/gatktodimsum.nf | 91 ++++ 2 files changed, 684 insertions(+), 104 deletions(-) create mode 100644 modules/local/gatk/gatktodimsum.nf diff --git a/log.txt b/log.txt index 4511556..38aa48b 100644 --- a/log.txt +++ b/log.txt @@ -1,124 +1,613 @@ -May-07 21:29:10.689 [main] DEBUG nextflow.cli.Launcher - $> nextflow run . -profile test,docker --fasta /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta --reading_frame 352-1383 --min_counts 2 --mutagenesis_type max_diff_to_wt --outdir ./results -May-07 21:29:10.864 [main] DEBUG nextflow.cli.CmdRun - N E X T F L O W ~ version 24.04.4 -May-07 21:29:10.887 [main] DEBUG nextflow.plugin.PluginsFacade - Setting up plugin manager > mode=prod; embedded=false; plugins-dir=/Users/benjaminwehnert/.nextflow/plugins; core-plugins: nf-amazon@2.5.3,nf-azure@1.6.1,nf-cloudcache@0.4.1,nf-codecommit@0.2.1,nf-console@1.1.3,nf-ga4gh@1.3.0,nf-google@1.13.2-patch1,nf-tower@1.9.1,nf-wave@1.4.2-patch1 -May-07 21:29:10.897 [main] INFO o.pf4j.DefaultPluginStatusProvider - Enabled plugins: [] -May-07 21:29:10.897 [main] INFO o.pf4j.DefaultPluginStatusProvider - Disabled plugins: [] -May-07 21:29:10.900 [main] INFO org.pf4j.DefaultPluginManager - PF4J version 3.12.0 in 'deployment' mode -May-07 21:29:10.914 [main] INFO org.pf4j.AbstractPluginManager - No plugins -May-07 21:29:11.769 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` -May-07 21:29:11.790 [main] DEBUG nextflow.config.ConfigBuilder - Found config local: /Users/benjaminwehnert/dmscore/nextflow.config -May-07 21:29:11.798 [main] DEBUG nextflow.config.ConfigBuilder - Parsing config file: /Users/benjaminwehnert/dmscore/nextflow.config -May-07 21:29:11.815 [main] DEBUG n.secret.LocalSecretsProvider - Secrets store: /Users/benjaminwehnert/.nextflow/secrets/store.json -May-07 21:29:11.818 [main] DEBUG nextflow.secret.SecretsLoader - Discovered secrets providers: [nextflow.secret.LocalSecretsProvider@6296474f] - activable => nextflow.secret.LocalSecretsProvider@6296474f -May-07 21:29:11.830 [main] DEBUG nextflow.config.ConfigBuilder - Applying config profile: `test,docker` -May-07 21:29:13.163 [main] ERROR nextflow.cli.Launcher - Unable to parse config file: '/Users/benjaminwehnert/dmscore/nextflow.config' +May-07 23:01:49.322 [main] DEBUG nextflow.cli.Launcher - $> nextflow run . -profile test,docker --fasta /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta --reading_frame 352-1383 --min_counts 2 --mutagenesis_type max_diff_to_wt --outdir ./results +May-07 23:01:49.500 [main] DEBUG nextflow.cli.CmdRun - N E X T F L O W ~ version 24.04.4 +May-07 23:01:49.524 [main] DEBUG nextflow.plugin.PluginsFacade - Setting up plugin manager > mode=prod; embedded=false; plugins-dir=/Users/benjaminwehnert/.nextflow/plugins; core-plugins: nf-amazon@2.5.3,nf-azure@1.6.1,nf-cloudcache@0.4.1,nf-codecommit@0.2.1,nf-console@1.1.3,nf-ga4gh@1.3.0,nf-google@1.13.2-patch1,nf-tower@1.9.1,nf-wave@1.4.2-patch1 +May-07 23:01:49.533 [main] INFO o.pf4j.DefaultPluginStatusProvider - Enabled plugins: [] +May-07 23:01:49.534 [main] INFO o.pf4j.DefaultPluginStatusProvider - Disabled plugins: [] +May-07 23:01:49.536 [main] INFO org.pf4j.DefaultPluginManager - PF4J version 3.12.0 in 'deployment' mode +May-07 23:01:49.549 [main] INFO org.pf4j.AbstractPluginManager - No plugins +May-07 23:01:50.385 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` +May-07 23:01:50.406 [main] DEBUG nextflow.config.ConfigBuilder - Found config local: /Users/benjaminwehnert/dmscore/nextflow.config +May-07 23:01:50.408 [main] DEBUG nextflow.config.ConfigBuilder - Parsing config file: /Users/benjaminwehnert/dmscore/nextflow.config +May-07 23:01:50.426 [main] DEBUG n.secret.LocalSecretsProvider - Secrets store: /Users/benjaminwehnert/.nextflow/secrets/store.json +May-07 23:01:50.428 [main] DEBUG nextflow.secret.SecretsLoader - Discovered secrets providers: [nextflow.secret.LocalSecretsProvider@169268a7] - activable => nextflow.secret.LocalSecretsProvider@169268a7 +May-07 23:01:50.438 [main] DEBUG nextflow.config.ConfigBuilder - Applying config profile: `test,docker` +May-07 23:01:51.909 [main] DEBUG nextflow.config.ConfigBuilder - Available config profiles: [bih, cfc_dev, uzl_omics, ifb_core, embl_hd, denbi_qbic, alice, mjolnir_globe, uppmax, giga, incliva, ilifu, ki_luria, uge, icr_alma, rosalind_uge, lugh, mccleary, unibe_ibu, vai, czbiohub_aws, jax, roslin, ccga_med, tes, scw, unc_longleaf, tigem, tubingen_apg, google, apollo, ipop_up, vsc_calcua, pdc_kth, googlels, ceci_nic5, humantechnopole, stjude, daisybio, eddie, medair, biowulf, apptainer, bi, bigpurple, adcra, cedars, pawsey_setonix, vsc_kul_uhasselt, pawsey_nimbus, ucl_myriad, utd_ganymede, charliecloud, seattlechildrens, icr_davros, ceres, arm, munin, rosalind, hasta, cfc, uzh, shu_bmrc, ebi_codon_slurm, ebc, ccga_dx, crick, ku_sund_danhead, marvin, shifter, biohpc_gen, mana, mamba, york_viking, unc_lccc, wehi, awsbatch, wustl_htcf, arcc, ceci_dragon2, imperial, maestro, software_license, cannon, genotoul, nci_gadi, abims, janelia, nu_genomics, googlebatch, oist, sahmri, kaust, alliance_canada, mpcdf, leicester, vsc_ugent, create, sage, cambridge, jex, podman, ebi_codon, cheaha, xanadu, nyu_hpc, test, marjorie, computerome, ucd_sonic, seg_globe, mssm, sanger, dkfz, bluebear, pasteur, einstein, ethz_euler, m3c, test_full, imb, ucl_cscluster, tuos_stanage, azurebatch, hki, seadragon, crukmi, csiro_petrichor, qmul_apocrita, wave, docker, engaging, gis, hypatia, psmn, eva, unity, cropdiversityhpc, nygc, fgcz, conda, crg, singularity, mpcdf_viper, pe2, self_hosted_runner, tufts, uw_hyak_pedslabs, binac2, debug, genouest, cbe, unsw_katana, gitpod, phoenix, seawulf, uod_hpc, fub_curta, uct_hpc, aws_tower, binac, fsu_draco] +May-07 23:01:51.958 [main] DEBUG nextflow.cli.CmdRun - Applied DSL=2 by global default +May-07 23:01:51.972 [main] DEBUG nextflow.cli.CmdRun - Launching `./main.nf` [modest_coulomb] DSL2 - revision: 84101fc51c +May-07 23:01:51.974 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins declared=[nf-schema@2.3.0] +May-07 23:01:51.974 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins default=[] +May-07 23:01:51.974 [main] DEBUG nextflow.plugin.PluginsFacade - Plugins resolved requirement=[nf-schema@2.3.0] +May-07 23:01:51.975 [main] DEBUG nextflow.plugin.PluginUpdater - Installing plugin nf-schema version: 2.3.0 +May-07 23:01:51.983 [main] INFO org.pf4j.AbstractPluginManager - Plugin 'nf-schema@2.3.0' resolved +May-07 23:01:51.983 [main] INFO org.pf4j.AbstractPluginManager - Start plugin 'nf-schema@2.3.0' +May-07 23:01:51.990 [main] DEBUG nextflow.plugin.BasePlugin - Plugin started nf-schema@2.3.0 +May-07 23:01:52.045 [main] DEBUG nextflow.Session - Session UUID: 2397b75e-3882-46d7-ba2c-8549f8a2b4a6 +May-07 23:01:52.045 [main] DEBUG nextflow.Session - Run name: modest_coulomb +May-07 23:01:52.046 [main] DEBUG nextflow.Session - Executor pool size: 8 +May-07 23:01:52.053 [main] DEBUG nextflow.file.FilePorter - File porter settings maxRetries=3; maxTransfers=50; pollTimeout=null +May-07 23:01:52.057 [main] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'FileTransfer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +May-07 23:01:52.077 [main] DEBUG nextflow.cli.CmdRun - + Version: 24.04.4 build 5917 + Created: 01-08-2024 07:05 UTC (09:05 CEST) + System: Mac OS X 15.0 + Runtime: Groovy 4.0.21 on OpenJDK 64-Bit Server VM 17.0.13+0 + Encoding: UTF-8 (UTF-8) + Process: 31358@MacBook-Air-von-Benjamin.local [127.0.0.1] + CPUs: 8 - Mem: 8 GB (94.6 MB) - Swap: 8 GB (737.5 MB) +May-07 23:01:52.088 [main] DEBUG nextflow.Session - Work-dir: /Users/benjaminwehnert/dmscore/work [Mac OS X] +May-07 23:01:52.088 [main] DEBUG nextflow.Session - Script base path does not exist or is not a directory: /Users/benjaminwehnert/dmscore/bin +May-07 23:01:52.104 [main] DEBUG nextflow.executor.ExecutorFactory - Extension executors providers=[] +May-07 23:01:52.123 [main] DEBUG nextflow.Session - Observer factory: DefaultObserverFactory +May-07 23:01:52.143 [main] DEBUG nextflow.Session - Observer factory: ValidationObserverFactory +May-07 23:01:52.174 [main] WARN nextflow.config.Manifest - Invalid config manifest attribute `contributors` +May-07 23:01:52.195 [main] DEBUG nextflow.cache.CacheFactory - Using Nextflow cache factory: nextflow.cache.DefaultCacheFactory +May-07 23:01:52.205 [main] DEBUG nextflow.util.CustomThreadPool - Creating default thread pool > poolSize: 9; maxThreads: 1000 +May-07 23:01:52.264 [main] DEBUG nextflow.Session - Session start +May-07 23:01:52.266 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow started -- trace file: /Users/benjaminwehnert/dmscore/results/pipeline_info/execution_trace_2025-05-07_23-01-50.txt +May-07 23:01:52.411 [main] DEBUG nextflow.script.ScriptRunner - > Launching execution +May-07 23:01:53.516 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +May-07 23:01:53.917 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema +May-07 23:01:53.918 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema +May-07 23:01:53.920 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +May-07 23:01:53.921 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema +May-07 23:01:54.131 [main] WARN nextflow.script.ScriptBinding - Access to undefined parameter `custom_codon_library` -- Initialise it to a default value eg. `params.custom_codon_library = some_value` +May-07 23:01:54.356 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryLog:paramsSummaryLog]; plugin Id: nf-schema +May-07 23:01:54.357 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [validateParameters:validateParameters]; plugin Id: nf-schema +May-07 23:01:54.359 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [paramsSummaryMap:paramsSummaryMap]; plugin Id: nf-schema +May-07 23:01:54.360 [main] DEBUG nextflow.script.IncludeDef - Loading included plugin extensions with names: [samplesheetToList:samplesheetToList]; plugin Id: nf-schema +May-07 23:01:54.574 [main] INFO nextflow.Nextflow - +------------------------------------------------------ + ,--./,-. + ___ __ __ __ ___ /,-._.--~' + |\ | |__ __ / ` / \ |__) |__ } { + | \| | \__, \__/ | \ |___ \`-._,-`-, + `._,._,' + nf-core/dmscore 1.0.0dev +------------------------------------------------------ +Input/output options + input : https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv + outdir : ./results + min_counts : 2 + mutagenesis_type : max_diff_to_wt - Compile failed for sources FixedSetSources[name='/groovy/script/Script7BD980DA7F8768B9E9654BF7A071C3F7']. Cause: org.codehaus.groovy.control.MultipleCompilationErrorsException: startup failed: - /groovy/script/Script7BD980DA7F8768B9E9654BF7A071C3F7: 13: Unexpected input: '{' @ line 13, column 9. - process { - ^ - - 1 error +Reference genome options + genome : R64-1-1 + fasta : /Users/benjaminwehnert/GID1A_SUNi_ref_small.fasta -ch.artecat.grengine.except.CompileException: Compile failed for sources FixedSetSources[name='/groovy/script/Script7BD980DA7F8768B9E9654BF7A071C3F7']. Cause: org.codehaus.groovy.control.MultipleCompilationErrorsException: startup failed: -/groovy/script/Script7BD980DA7F8768B9E9654BF7A071C3F7: 13: Unexpected input: '{' @ line 13, column 9. - process { - ^ +Institutional config options + config_profile_name : Test profile + config_profile_description: Minimal test dataset to check pipeline function -1 error +Generic options + trace_report_suffix : 2025-05-07_23-01-50 - at ch.artecat.grengine.code.groovy.DefaultGroovyCompiler.compile(DefaultGroovyCompiler.java:260) - at ch.artecat.grengine.load.DefaultTopCodeCache.getUpToDateCode(DefaultTopCodeCache.java:80) - at ch.artecat.grengine.load.LayeredClassLoader.getTopLoader(LayeredClassLoader.java:251) - at ch.artecat.grengine.load.LayeredClassLoader.loadMainClass(LayeredClassLoader.java:202) - at ch.artecat.grengine.engine.LayeredEngine.loadMainClass(LayeredEngine.java:198) - at ch.artecat.grengine.Grengine.loadMainClass(Grengine.java:679) - at ch.artecat.grengine.BaseGrengine.load(BaseGrengine.java:538) - at ch.artecat.grengine.BaseGrengine.load(BaseGrengine.java:387) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.selectMethod(IndyInterface.java:355) +Core Nextflow options + runName : modest_coulomb + containerEngine : docker + launchDir : /Users/benjaminwehnert/dmscore + workDir : /Users/benjaminwehnert/dmscore/work + projectDir : /Users/benjaminwehnert/dmscore + userName : benjaminwehnert + profile : test,docker + configFiles : /Users/benjaminwehnert/dmscore/nextflow.config + +!! Only displaying parameters that differ from the pipeline defaults !! +------------------------------------------------------ +* The nf-core framework + https://doi.org/10.1038/s41587-020-0439-x + +* Software dependencies + https://github.com/nf-core/dmscore/blob/master/CITATIONS.md + +May-07 23:01:54.576 [main] DEBUG n.validation.ValidationExtension - Starting parameters validation +May-07 23:01:54.920 [main] DEBUG nextflow.validation.SchemaEvaluator - Started validating /BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv +May-07 23:01:55.945 [main] DEBUG nextflow.validation.SchemaEvaluator - Validation of file 'https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/samplesheet_qc_only.csv' passed! +May-07 23:01:56.095 [main] DEBUG n.v.FormatDirectoryPathEvaluator - Cloud blob storage paths are not supported by 'FormatDirectoryPathEvaluator': 's3://ngi-igenomes/igenomes/' +May-07 23:01:56.099 [main] DEBUG n.validation.ValidationExtension - Finishing parameters validation +May-07 23:01:56.178 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:FASTQC +May-07 23:01:56.182 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:FASTQC` matches process NFCORE_DMSCORE:DMSCORE:FASTQC +May-07 23:01:56.196 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-07 23:01:56.196 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-07 23:01:56.202 [main] DEBUG nextflow.executor.Executor - [warm up] executor > local +May-07 23:01:56.206 [main] DEBUG n.processor.LocalPollingMonitor - Creating local task monitor for executor 'local' > cpus=8; memory=8 GB; capacity=8; pollInterval=100ms; dumpInterval=5m +May-07 23:01:56.209 [main] DEBUG n.processor.TaskPollingMonitor - >>> barrier register (monitor: local) +May-07 23:01:56.359 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:MULTIQC +May-07 23:01:56.360 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:MULTIQC` matches process NFCORE_DMSCORE:DMSCORE:MULTIQC +May-07 23:01:56.362 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-07 23:01:56.363 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-07 23:01:56.372 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BWA_INDEX +May-07 23:01:56.373 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:BWA_INDEX` matches process NFCORE_DMSCORE:DMSCORE:BWA_INDEX +May-07 23:01:56.375 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-07 23:01:56.375 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-07 23:01:56.391 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_high` matches labels `process_high` for process with name NFCORE_DMSCORE:DMSCORE:BWA_MEM +May-07 23:01:56.391 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:BWA_MEM` matches process NFCORE_DMSCORE:DMSCORE:BWA_MEM +May-07 23:01:56.394 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-07 23:01:56.395 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-07 23:01:56.404 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS +May-07 23:01:56.406 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:BAMFILTER_DMS` matches process NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS +May-07 23:01:56.409 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-07 23:01:56.409 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-07 23:01:56.418 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_medium` matches labels `process_medium` for process with name NFCORE_DMSCORE:DMSCORE:PREMERGE +May-07 23:01:56.418 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:PREMERGE` matches process NFCORE_DMSCORE:DMSCORE:PREMERGE +May-07 23:01:56.420 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-07 23:01:56.420 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-07 23:01:56.429 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_high` matches labels `process_high` for process with name NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS +May-07 23:01:56.430 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:GATK_SATURATIONMUTAGENESIS` matches process NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS +May-07 23:01:56.433 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-07 23:01:56.433 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-07 23:01:56.443 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ +May-07 23:01:56.443 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:DMSANALYSIS_AASEQ` matches process NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ +May-07 23:01:56.446 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-07 23:01:56.446 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-07 23:01:56.458 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_POSSIBLE_MUTATIONS +May-07 23:01:56.459 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:DMSANALYSIS_POSSIBLE_MUTATIONS` matches process NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_POSSIBLE_MUTATIONS +May-07 23:01:56.461 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-07 23:01:56.461 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-07 23:01:56.480 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withLabel:process_single` matches labels `process_single` for process with name NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_PROCESS_GATK +May-07 23:01:56.481 [main] DEBUG nextflow.script.ProcessConfig - Config settings `withName:DMSANALYSIS_PROCESS_GATK` matches process NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_PROCESS_GATK +May-07 23:01:56.484 [main] DEBUG nextflow.executor.ExecutorFactory - << taskConfig executor: null +May-07 23:01:56.484 [main] DEBUG nextflow.executor.ExecutorFactory - >> processorType: 'local' +May-07 23:01:56.499 [main] DEBUG nextflow.Session - Config process names validation disabled as requested +May-07 23:01:56.500 [main] DEBUG nextflow.Session - Igniting dataflow network (43) +May-07 23:01:56.508 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:FASTQC +May-07 23:01:56.508 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:MULTIQC +May-07 23:01:56.509 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX +May-07 23:01:56.509 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BWA_MEM +May-07 23:01:56.511 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS +May-07 23:01:56.512 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:PREMERGE +May-07 23:01:56.512 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS +May-07 23:01:56.512 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ +May-07 23:01:56.512 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_POSSIBLE_MUTATIONS +May-07 23:01:56.512 [main] DEBUG nextflow.processor.TaskProcessor - Starting process > NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_PROCESS_GATK +May-07 23:01:56.514 [main] DEBUG nextflow.script.ScriptRunner - Parsed script files: + Script_e5f965e09aa3641b: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/bamprocessing/premerge.nf + Script_113f045ba19c2c46: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/fastqc/main.nf + Script_4f612cfd8c52e8cd: /Users/benjaminwehnert/dmscore/./subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nextflow_pipeline/main.nf + Script_215a636da7ab24a2: /Users/benjaminwehnert/dmscore/./workflows/../subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfschema_plugin/main.nf + Script_68606fccdecc54d9: /Users/benjaminwehnert/dmscore/./subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf + Script_0f3077e98f6a93f6: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/mem/main.nf + Script_5c4e8d4051efa81e: /Users/benjaminwehnert/dmscore/./subworkflows/local/utils_nfcore_dmscore_pipeline/../../nf-core/utils_nfcore_pipeline/main.nf + Script_3bf8120d0b5bcde1: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/dmsanalysis/possiblemutations.nf + Script_d65a0ba0a319d7db: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/dmsanalysis/aaseq.nf + Script_4ff7366a79e0ef06: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/bamprocessing/bamfilteringdms.nf + Script_c568aae5b239c1c5: /Users/benjaminwehnert/dmscore/main.nf + Script_09ccfa79b2802f41: /Users/benjaminwehnert/dmscore/./workflows/dmscore.nf + Script_4955288afd8ca61e: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/multiqc/main.nf + Script_a1878766b1a6b241: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/dmsanalysis/processgatk.nf + Script_37dcd664b2773148: /Users/benjaminwehnert/dmscore/./workflows/../modules/local/gatk/saturationmutagenesis.nf + Script_25aa31c18d513c61: /Users/benjaminwehnert/dmscore/./workflows/../modules/nf-core/bwa/index/main.nf +May-07 23:01:56.514 [main] DEBUG nextflow.script.ScriptRunner - > Awaiting termination +May-07 23:01:56.514 [main] DEBUG nextflow.Session - Session await +May-07 23:01:56.605 [Actor Thread 1] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.001 s; external sort time: 0.011 s; total time: 0.012 s +May-07 23:01:56.605 [Actor Thread 2] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 1; slices: 1; internal sort time: 0.001 s; external sort time: 0.011 s; total time: 0.012 s +May-07 23:01:56.611 [Actor Thread 2] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/4640128d9cfd4a45636425a4f43db374 +May-07 23:01:56.611 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/04a58cc29ef1c76e5af4e2fb20a13ad7 +May-07 23:01:56.619 [Actor Thread 1] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-1651345332501275985 +May-07 23:01:56.619 [Actor Thread 2] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-12321695686053957190 +May-07 23:01:56.623 [Actor Thread 3] DEBUG nextflow.util.HashBuilder - [WARN] Unknown hashing type: class Script_09ccfa79b2802f41$_runScript_closure1$_closure26 +May-07 23:01:56.624 [Actor Thread 14] DEBUG nextflow.util.HashBuilder - Unable to get file attributes file: /NULL -- Cause: java.nio.file.NoSuchFileException: /NULL +May-07 23:01:56.697 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-07 23:01:56.698 [Task submitter] INFO nextflow.Session - [d4/e141d7] Submitted process > NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (amino_acid_sequence) +May-07 23:01:56.730 [FileTransfer-2] DEBUG nextflow.file.FilePorter - Copying foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq to work dir: /Users/benjaminwehnert/dmscore/work/stage-2397b75e-3882-46d7-ba2c-8549f8a2b4a6/6a/8815724cee5e2e4e17200e98e8c0ad/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq +May-07 23:01:56.730 [FileTransfer-1] DEBUG nextflow.file.FilePorter - Copying foreign file https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq to work dir: /Users/benjaminwehnert/dmscore/work/stage-2397b75e-3882-46d7-ba2c-8549f8a2b4a6/40/dfe72a8fa5ce1b038fdf1aa1fb4749/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq +May-07 23:01:58.744 [Actor Thread 15] INFO nextflow.file.FilePorter - Staging foreign file: https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R1_50k.fastq +May-07 23:02:00.581 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 3; name: NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (amino_acid_sequence); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/d4/e141d76a7c3b4130080bcdf8a831a3] +May-07 23:02:00.585 [Task monitor] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'TaskFinalizer' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +May-07 23:02:00.612 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-07 23:02:00.613 [Task submitter] INFO nextflow.Session - [49/349fd9] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta) +May-07 23:02:00.663 [TaskFinalizer-1] DEBUG nextflow.util.ThreadPoolBuilder - Creating thread pool 'PublishDir' minSize=10; maxSize=24; workQueue=LinkedBlockingQueue[10000]; allowCoreThreadTimeout=false +May-07 23:02:00.756 [Actor Thread 15] INFO nextflow.file.FilePorter - Staging foreign file: https://raw.githubusercontent.com/BenjaminWehnert1008/test-datasets/dmsqc/dmsqc/pMS190_GID1A_SUNi_S2_1_R2_50k.fastq +May-07 23:02:00.757 [Actor Thread 4] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process +May-07 23:02:00.763 [Actor Thread 4] DEBUG nextflow.processor.TaskContext - Failed to serialize delegate map items: [ + 'meta':[Script_09ccfa79b2802f41$_runScript_closure1$_closure26] = + 'pos_range':[java.lang.String] = 352-1383 + '$':[java.lang.Boolean] = true + 'wt_seq':[nextflow.processor.TaskPath] = GID1A_SUNi_ref_small.fasta + 'script':[nextflow.processor.TaskPath] = aa_seq.R + 'task':[nextflow.processor.TaskConfig] = [container:community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa, withName:PREMERGE:[publishDir:[path:./results/intermediate_files/bam_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure9$_closure21@26dc7ffc]], memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure15@13f9d575, withName:FASTQC:[ext:[args:--quiet], containerOptions:], withLabel:error_retry:[errorStrategy:retry, maxRetries:2], when:nextflow.script.TaskClosure@264d7648, withLabel:process_high:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure23@1b36f3f3, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure24@5e504cf6, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure25@35f6834], withLabel:error_ignore:[errorStrategy:ignore], resourceLimits:[cpus:4, memory:8.GB, time:1.h], withName:DMSANALYSIS_AASEQ:[publishDir:[path:./results/intermediate_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure13$_closure25@4eeda121]], withLabel:process_high_memory:[memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure11$_closure27@62e86a64], withName:BAMFILTER_DMS:[publishDir:[path:./results/intermediate_files/bam_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure8$_closure20@3e03bd33]], publishDir:[[path:./results/intermediate_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure13$_closure25@4eeda121]], withName:BWA_MEM:[publishDir:[path:./results/intermediate_files/bam_files/bwa/mem, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure7$_closure19@46e56c0f]], executor:local, stub:nextflow.script.TaskClosure@6522395b, conda:null/environment.yml, withName:GATK_SATURATIONMUTAGENESIS:[publishDir:[path:./results/intermediate_files/gatk, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure10$_closure22@4438d4d1]], cacheable:true, withLabel:process_low:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure17@67f11340, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure18@a998ea5, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure19@7e85964c], withLabel:process_medium:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure20@7c995b11, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure21@131d3cd1, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure22@55b774b1], tag:amino_acid_sequence, withName:MULTIQC:[ext:[args:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure5$_closure15@7d2bfbd], publishDir:[path:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure5$_closure16@31a52d85, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure5$_closure17@4ba464d4]], workDir:/Users/benjaminwehnert/dmscore/work/d4/e141d76a7c3b4130080bcdf8a831a3, exitStatus:0, ext:[:], withLabel:process_single:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure14@255893ed, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure15@13f9d575, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure16@37e5efac], withName:BWA_INDEX:[publishDir:[path:./results/intermediate_files/bam_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure6$_closure18@2f3435d0]], process:NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ, debug:false, cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure14@255893ed, index:1, label:[process_single], withLabel:process_long:[time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure10$_closure26@475e6626], maxRetries:1, maxErrors:-1, shell:bash + +set -e # Exit if a tool returns a non-zero status/exit code +set -u # Treat unset variables and parameters as an error +set -o pipefail # Returns the status of the last command to exit with a non-zero status or zero if all successfully execute +set -C # No clobber - prevent output redirection from overwriting files. +, withName:DMSANALYSIS_POSSIBLE_MUTATIONS:[publishDir:[path:./results/intermediate_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure11$_closure23@267852db]], name:NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_AASEQ (amino_acid_sequence), containerOptions:-u $(id -u):$(id -g), errorStrategy:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure5@3e785137, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure16@37e5efac, withName:DMSANALYSIS_PROCESS_GATK:[publishDir:[path:./results/intermediate_files/processed_gatk_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure14$_closure26@30ec799d]], hash:d4e141d76a7c3b4130080bcdf8a831a3] +] +com.esotericsoftware.kryo.KryoException: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue +Serialization trace: +queue (org.codehaus.groovy.util.ReferenceManager$CallBackedManager) +manager (org.codehaus.groovy.util.ReferenceManager$1) +manager (org.codehaus.groovy.util.ReferenceBundle) +bundle (org.codehaus.groovy.reflection.CachedClass$4) +cachedSuperClass (org.codehaus.groovy.reflection.stdclasses.ObjectCachedClass) +cachedClass (org.codehaus.groovy.reflection.CachedMethod) +allMethods (groovy.lang.MetaClassImpl) +delegate (groovy.runtime.metaclass.NextflowDelegatingMetaClass) +metaClass (groovyx.gpars.dataflow.DataflowVariable) +first (groovyx.gpars.dataflow.stream.DataflowStream) +asyncHead (groovyx.gpars.dataflow.stream.DataflowStreamReadAdapter) +source (nextflow.extension.MapOp) +owner (nextflow.extension.MapOp$_apply_closure1) +code (groovyx.gpars.dataflow.operator.DataflowOperatorActor) +actor (groovyx.gpars.dataflow.operator.DataflowOperator) +allOperators (nextflow.Session) +session (nextflow.validation.ValidationExtension) +target (nextflow.script.FunctionDef) +definitions (nextflow.script.ScriptMeta) +meta (nextflow.script.ScriptBinding) +binding (Script_09ccfa79b2802f41) +delegate (Script_09ccfa79b2802f41$_runScript_closure1) +delegate (Script_09ccfa79b2802f41$_runScript_closure1$_closure26) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:82) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:82) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:22) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:82) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:22) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:95) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:21) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:95) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:21) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigBase.includeConfig(ConfigBase.groovy:112) + at nextflow.util.KryoHelper.serialize(SerializationHelper.groovy:166) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.processor.TaskContext.serialize(TaskContext.groovy:198) + at nextflow.cache.CacheDB.writeTaskEntry0(CacheDB.groovy:148) at java.base/jdk.internal.reflect.NativeMethodAccessorImpl.invoke0(Native Method) at java.base/jdk.internal.reflect.NativeMethodAccessorImpl.invoke(NativeMethodAccessorImpl.java:77) at java.base/jdk.internal.reflect.DelegatingMethodAccessorImpl.invoke(DelegatingMethodAccessorImpl.java:43) at java.base/java.lang.reflect.Method.invoke(Method.java:569) at org.codehaus.groovy.reflection.CachedMethod.invoke(CachedMethod.java:343) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.selectMethod(IndyInterface.java:355) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigParser$_parse_closure5.doCall(ConfigParser.groovy:457) - at jdk.internal.reflect.GeneratedMethodAccessor19.invoke(Unknown Source) - at java.base/jdk.internal.reflect.DelegatingMethodAccessorImpl.invoke(DelegatingMethodAccessorImpl.java:43) - at java.base/java.lang.reflect.Method.invoke(Method.java:569) - at org.codehaus.groovy.reflection.CachedMethod.invoke(CachedMethod.java:343) - at org.codehaus.groovy.runtime.metaclass.ClosureMetaMethod.invoke(ClosureMetaMethod.java:88) - at groovy.lang.ExpandoMetaClass.invokeMethod(ExpandoMetaClass.java:1140) + at groovy.lang.MetaMethod.doMethodInvoke(MetaMethod.java:328) + at groovy.lang.MetaClassImpl.doInvokeMethod(MetaClassImpl.java:1333) + at groovy.lang.MetaClassImpl.invokeMethod(MetaClassImpl.java:1088) at groovy.lang.MetaClassImpl.invokeMethod(MetaClassImpl.java:1007) - at groovy.lang.DelegatingMetaClass.invokeMethod(DelegatingMetaClass.java:165) + at org.codehaus.groovy.runtime.InvokerHelper.invokePogoMethod(InvokerHelper.java:645) + at org.codehaus.groovy.runtime.InvokerHelper.invokeMethod(InvokerHelper.java:628) + at org.codehaus.groovy.runtime.InvokerHelper.invokeMethodSafe(InvokerHelper.java:82) + at nextflow.cache.CacheDB$_putTaskAsync_closure1.doCall(CacheDB.groovy:157) + at nextflow.cache.CacheDB$_putTaskAsync_closure1.call(CacheDB.groovy) + at groovyx.gpars.agent.AgentBase.onMessage(AgentBase.java:102) + at groovyx.gpars.agent.Agent.handleMessage(Agent.java:84) + at groovyx.gpars.agent.AgentCore$1.handleMessage(AgentCore.java:48) + at groovyx.gpars.util.AsyncMessagingCore.run(AsyncMessagingCore.java:132) + at java.base/java.util.concurrent.ThreadPoolExecutor.runWorker(ThreadPoolExecutor.java:1136) + at java.base/java.util.concurrent.ThreadPoolExecutor$Worker.run(ThreadPoolExecutor.java:635) + at java.base/java.lang.Thread.run(Thread.java:840) +Caused by: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue + at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:48) + at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:26) + at com.esotericsoftware.kryo.Kryo.newDefaultSerializer(Kryo.java:351) + at com.esotericsoftware.kryo.Kryo.getDefaultSerializer(Kryo.java:344) + at com.esotericsoftware.kryo.util.DefaultClassResolver.registerImplicit(DefaultClassResolver.java:56) + at com.esotericsoftware.kryo.Kryo.getRegistration(Kryo.java:461) + at com.esotericsoftware.kryo.util.DefaultClassResolver.writeClass(DefaultClassResolver.java:79) + at com.esotericsoftware.kryo.Kryo.writeClass(Kryo.java:488) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:57) + ... 106 common frames omitted +Caused by: java.lang.reflect.InvocationTargetException: null + at jdk.internal.reflect.GeneratedConstructorAccessor39.newInstance(Unknown Source) + at java.base/jdk.internal.reflect.DelegatingConstructorAccessorImpl.newInstance(DelegatingConstructorAccessorImpl.java:45) + at java.base/java.lang.reflect.Constructor.newInstanceWithCaller(Constructor.java:500) + at java.base/java.lang.reflect.Constructor.newInstance(Constructor.java:481) + at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:35) + ... 114 common frames omitted +Caused by: java.lang.reflect.InaccessibleObjectException: Unable to make field private final java.lang.ref.ReferenceQueue$Lock java.lang.ref.ReferenceQueue.lock accessible: module java.base does not "opens java.lang.ref" to unnamed module @7b02881e + at java.base/java.lang.reflect.AccessibleObject.checkCanSetAccessible(AccessibleObject.java:354) + at java.base/java.lang.reflect.AccessibleObject.checkCanSetAccessible(AccessibleObject.java:297) + at java.base/java.lang.reflect.Field.checkCanSetAccessible(Field.java:178) + at java.base/java.lang.reflect.Field.setAccessible(Field.java:172) + at com.esotericsoftware.kryo.serializers.FieldSerializer.buildValidFields(FieldSerializer.java:282) + at com.esotericsoftware.kryo.serializers.FieldSerializer.rebuildCachedFields(FieldSerializer.java:217) + at com.esotericsoftware.kryo.serializers.FieldSerializer.rebuildCachedFields(FieldSerializer.java:156) + at com.esotericsoftware.kryo.serializers.FieldSerializer.(FieldSerializer.java:133) + ... 119 common frames omitted +May-07 23:02:02.378 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 1; name: NFCORE_DMSCORE:DMSCORE:BWA_INDEX (GID1A_SUNi_ref_small.fasta); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/49/349fd93ff7dd98e92e426a30457fb9] +May-07 23:02:02.387 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-07 23:02:02.388 [Task submitter] INFO nextflow.Session - [36/b138e8] Submitted process > NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_POSSIBLE_MUTATIONS (table /w all possible variants) +May-07 23:02:02.422 [Actor Thread 8] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process +May-07 23:02:13.067 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 2; name: NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_POSSIBLE_MUTATIONS (table /w all possible variants); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/36/b138e8e27f7423eea98b17aaf74451] +May-07 23:02:13.085 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-07 23:02:13.085 [Task submitter] INFO nextflow.Session - [67/c2a06b] Submitted process > NFCORE_DMSCORE:DMSCORE:BWA_MEM (gid1a_1_quality_1_pe) +May-07 23:02:13.133 [Actor Thread 6] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process +May-07 23:02:13.136 [Actor Thread 6] DEBUG nextflow.processor.TaskContext - Failed to serialize delegate map items: [ + 'meta':[Script_09ccfa79b2802f41$_runScript_closure1$_closure26] = + 'pos_range':[java.lang.String] = 352-1383 + 'mutagenesis_type':[java.lang.String] = max_diff_to_wt + '$':[java.lang.Boolean] = true + 'wt_seq':[nextflow.processor.TaskPath] = GID1A_SUNi_ref_small.fasta + 'custom_codon_library':[nextflow.processor.TaskPath] = NULL + 'script':[nextflow.processor.TaskPath] = possible_mutations.R + 'task':[nextflow.processor.TaskConfig] = [container:community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa, withName:PREMERGE:[publishDir:[path:./results/intermediate_files/bam_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure9$_closure21@26dc7ffc]], memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure15@13f9d575, withName:FASTQC:[ext:[args:--quiet], containerOptions:], withLabel:error_retry:[errorStrategy:retry, maxRetries:2], when:nextflow.script.TaskClosure@5f4899a5, withLabel:process_high:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure23@1b36f3f3, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure24@5e504cf6, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure9$_closure25@35f6834], withLabel:error_ignore:[errorStrategy:ignore], resourceLimits:[cpus:4, memory:8.GB, time:1.h], withName:DMSANALYSIS_AASEQ:[publishDir:[path:./results/intermediate_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure13$_closure25@4eeda121]], withLabel:process_high_memory:[memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure11$_closure27@62e86a64], withName:BAMFILTER_DMS:[publishDir:[path:./results/intermediate_files/bam_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure8$_closure20@3e03bd33]], publishDir:[[path:./results/intermediate_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure11$_closure23@267852db]], withName:BWA_MEM:[publishDir:[path:./results/intermediate_files/bam_files/bwa/mem, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure7$_closure19@46e56c0f]], executor:local, stub:nextflow.script.TaskClosure@7069093e, conda:null/environment.yml, withName:GATK_SATURATIONMUTAGENESIS:[publishDir:[path:./results/intermediate_files/gatk, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure10$_closure22@4438d4d1]], cacheable:true, withLabel:process_low:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure17@67f11340, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure18@a998ea5, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure7$_closure19@7e85964c], withLabel:process_medium:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure20@7c995b11, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure21@131d3cd1, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure8$_closure22@55b774b1], tag:table /w all possible variants, withName:MULTIQC:[ext:[args:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure5$_closure15@7d2bfbd], publishDir:[path:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure5$_closure16@31a52d85, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure5$_closure17@4ba464d4]], workDir:/Users/benjaminwehnert/dmscore/work/36/b138e8e27f7423eea98b17aaf74451, exitStatus:0, ext:[:], withLabel:process_single:[cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure14@255893ed, memory:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure15@13f9d575, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure16@37e5efac], withName:BWA_INDEX:[publishDir:[path:./results/intermediate_files/bam_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure6$_closure18@2f3435d0]], process:NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_POSSIBLE_MUTATIONS, debug:false, cpus:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure14@255893ed, index:1, label:[process_single], withLabel:process_long:[time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure10$_closure26@475e6626], maxRetries:1, maxErrors:-1, shell:bash + +set -e # Exit if a tool returns a non-zero status/exit code +set -u # Treat unset variables and parameters as an error +set -o pipefail # Returns the status of the last command to exit with a non-zero status or zero if all successfully execute +set -C # No clobber - prevent output redirection from overwriting files. +, withName:DMSANALYSIS_POSSIBLE_MUTATIONS:[publishDir:[path:./results/intermediate_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure11$_closure23@267852db]], name:NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_POSSIBLE_MUTATIONS (table /w all possible variants), containerOptions:-u $(id -u):$(id -g), errorStrategy:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure5@3e785137, time:Script126E8BAA3BC9692B58F2A1C965A1D472$_run_closure1$_closure6$_closure16@37e5efac, withName:DMSANALYSIS_PROCESS_GATK:[publishDir:[path:./results/intermediate_files/processed_gatk_files, mode:copy, saveAs:ScriptE8646A4B8FFA7429020F836DC9CB8146$_run_closure1$_closure14$_closure26@30ec799d]], hash:36b138e8e27f7423eea98b17aaf74451] +] +com.esotericsoftware.kryo.KryoException: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue +Serialization trace: +queue (org.codehaus.groovy.util.ReferenceManager$CallBackedManager) +manager (org.codehaus.groovy.util.ReferenceManager$1) +manager (org.codehaus.groovy.util.ReferenceBundle) +bundle (org.codehaus.groovy.reflection.CachedClass$4) +cachedSuperClass (org.codehaus.groovy.reflection.stdclasses.ObjectCachedClass) +cachedClass (org.codehaus.groovy.reflection.CachedMethod) +allMethods (groovy.lang.MetaClassImpl) +delegate (groovy.runtime.metaclass.NextflowDelegatingMetaClass) +metaClass (groovyx.gpars.dataflow.DataflowVariable) +first (groovyx.gpars.dataflow.stream.DataflowStream) +asyncHead (groovyx.gpars.dataflow.stream.DataflowStreamReadAdapter) +source (nextflow.extension.MapOp) +owner (nextflow.extension.MapOp$_apply_closure1) +code (groovyx.gpars.dataflow.operator.DataflowOperatorActor) +actor (groovyx.gpars.dataflow.operator.DataflowOperator) +allOperators (nextflow.Session) +session (nextflow.validation.ValidationExtension) +target (nextflow.script.FunctionDef) +definitions (nextflow.script.ScriptMeta) +meta (nextflow.script.ScriptBinding) +binding (Script_09ccfa79b2802f41) +delegate (Script_09ccfa79b2802f41$_runScript_closure1) +delegate (Script_09ccfa79b2802f41$_runScript_closure1$_closure26) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:82) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:82) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:22) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:82) + at com.esotericsoftware.kryo.serializers.CollectionSerializer.write(CollectionSerializer.java:22) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:95) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:21) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeObject(Kryo.java:523) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:61) + at com.esotericsoftware.kryo.serializers.FieldSerializer.write(FieldSerializer.java:495) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:95) + at com.esotericsoftware.kryo.serializers.MapSerializer.write(MapSerializer.java:21) + at com.esotericsoftware.kryo.Kryo.writeClassAndObject(Kryo.java:599) at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at _nf_config_af5e21ee.run(_nf_config_af5e21ee:288) + at nextflow.util.KryoHelper.serialize(SerializationHelper.groovy:166) + at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) + at nextflow.processor.TaskContext.serialize(TaskContext.groovy:198) + at nextflow.cache.CacheDB.writeTaskEntry0(CacheDB.groovy:148) at java.base/jdk.internal.reflect.NativeMethodAccessorImpl.invoke0(Native Method) at java.base/jdk.internal.reflect.NativeMethodAccessorImpl.invoke(NativeMethodAccessorImpl.java:77) at java.base/jdk.internal.reflect.DelegatingMethodAccessorImpl.invoke(DelegatingMethodAccessorImpl.java:43) at java.base/java.lang.reflect.Method.invoke(Method.java:569) at org.codehaus.groovy.reflection.CachedMethod.invoke(CachedMethod.java:343) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigParser$_parse_closure5.doCall(ConfigParser.groovy:457) - at jdk.internal.reflect.GeneratedMethodAccessor19.invoke(Unknown Source) - at java.base/jdk.internal.reflect.DelegatingMethodAccessorImpl.invoke(DelegatingMethodAccessorImpl.java:43) - at java.base/java.lang.reflect.Method.invoke(Method.java:569) - at org.codehaus.groovy.reflection.CachedMethod.invoke(CachedMethod.java:343) - at org.codehaus.groovy.runtime.metaclass.ClosureMetaMethod.invoke(ClosureMetaMethod.java:88) - at groovy.lang.ExpandoMetaClass.invokeMethod(ExpandoMetaClass.java:1140) + at groovy.lang.MetaMethod.doMethodInvoke(MetaMethod.java:328) + at groovy.lang.MetaClassImpl.doInvokeMethod(MetaClassImpl.java:1333) + at groovy.lang.MetaClassImpl.invokeMethod(MetaClassImpl.java:1088) at groovy.lang.MetaClassImpl.invokeMethod(MetaClassImpl.java:1007) - at groovy.lang.DelegatingMetaClass.invokeMethod(DelegatingMetaClass.java:165) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.selectMethod(IndyInterface.java:355) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigParser.parse(ConfigParser.groovy:486) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigParser.parse(ConfigParser.groovy:312) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigBuilder.parse0(ConfigBuilder.groovy:439) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigBuilder.merge0(ConfigBuilder.groovy:425) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigBuilder.buildConfig0(ConfigBuilder.groovy:375) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigBuilder.buildGivenFiles(ConfigBuilder.groovy:320) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigBuilder.buildConfigObject(ConfigBuilder.groovy:814) - at org.codehaus.groovy.vmplugin.v8.IndyInterface.fromCache(IndyInterface.java:321) - at nextflow.config.ConfigBuilder.build(ConfigBuilder.groovy:827) - at nextflow.cli.CmdRun.run(CmdRun.groovy:328) - at nextflow.cli.Launcher.run(Launcher.groovy:503) - at nextflow.cli.Launcher.main(Launcher.groovy:657) -Caused by: org.codehaus.groovy.control.MultipleCompilationErrorsException: startup failed: -/groovy/script/Script7BD980DA7F8768B9E9654BF7A071C3F7: 13: Unexpected input: '{' @ line 13, column 9. - process { - ^ + at org.codehaus.groovy.runtime.InvokerHelper.invokePogoMethod(InvokerHelper.java:645) + at org.codehaus.groovy.runtime.InvokerHelper.invokeMethod(InvokerHelper.java:628) + at org.codehaus.groovy.runtime.InvokerHelper.invokeMethodSafe(InvokerHelper.java:82) + at nextflow.cache.CacheDB$_putTaskAsync_closure1.doCall(CacheDB.groovy:157) + at nextflow.cache.CacheDB$_putTaskAsync_closure1.call(CacheDB.groovy) + at groovyx.gpars.agent.AgentBase.onMessage(AgentBase.java:102) + at groovyx.gpars.agent.Agent.handleMessage(Agent.java:84) + at groovyx.gpars.agent.AgentCore$1.handleMessage(AgentCore.java:48) + at groovyx.gpars.util.AsyncMessagingCore.run(AsyncMessagingCore.java:132) + at java.base/java.util.concurrent.ThreadPoolExecutor.runWorker(ThreadPoolExecutor.java:1136) + at java.base/java.util.concurrent.ThreadPoolExecutor$Worker.run(ThreadPoolExecutor.java:635) + at java.base/java.lang.Thread.run(Thread.java:840) +Caused by: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue + at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:48) + at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:26) + at com.esotericsoftware.kryo.Kryo.newDefaultSerializer(Kryo.java:351) + at com.esotericsoftware.kryo.Kryo.getDefaultSerializer(Kryo.java:344) + at com.esotericsoftware.kryo.util.DefaultClassResolver.registerImplicit(DefaultClassResolver.java:56) + at com.esotericsoftware.kryo.Kryo.getRegistration(Kryo.java:461) + at com.esotericsoftware.kryo.util.DefaultClassResolver.writeClass(DefaultClassResolver.java:79) + at com.esotericsoftware.kryo.Kryo.writeClass(Kryo.java:488) + at com.esotericsoftware.kryo.serializers.ObjectField.write(ObjectField.java:57) + ... 106 common frames omitted +Caused by: java.lang.reflect.InvocationTargetException: null + at jdk.internal.reflect.GeneratedConstructorAccessor39.newInstance(Unknown Source) + at java.base/jdk.internal.reflect.DelegatingConstructorAccessorImpl.newInstance(DelegatingConstructorAccessorImpl.java:45) + at java.base/java.lang.reflect.Constructor.newInstanceWithCaller(Constructor.java:500) + at java.base/java.lang.reflect.Constructor.newInstance(Constructor.java:481) + at com.esotericsoftware.kryo.factories.ReflectionSerializerFactory.makeSerializer(ReflectionSerializerFactory.java:35) + ... 114 common frames omitted +Caused by: java.lang.reflect.InaccessibleObjectException: Unable to make field private final java.lang.ref.ReferenceQueue$Lock java.lang.ref.ReferenceQueue.lock accessible: module java.base does not "opens java.lang.ref" to unnamed module @7b02881e + at java.base/java.lang.reflect.AccessibleObject.checkCanSetAccessible(AccessibleObject.java:354) + at java.base/java.lang.reflect.AccessibleObject.checkCanSetAccessible(AccessibleObject.java:297) + at java.base/java.lang.reflect.Field.checkCanSetAccessible(Field.java:178) + at java.base/java.lang.reflect.Field.setAccessible(Field.java:172) + at com.esotericsoftware.kryo.serializers.FieldSerializer.buildValidFields(FieldSerializer.java:282) + at com.esotericsoftware.kryo.serializers.FieldSerializer.rebuildCachedFields(FieldSerializer.java:217) + at com.esotericsoftware.kryo.serializers.FieldSerializer.rebuildCachedFields(FieldSerializer.java:156) + at com.esotericsoftware.kryo.serializers.FieldSerializer.(FieldSerializer.java:133) + ... 119 common frames omitted +May-07 23:02:17.570 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 5; name: NFCORE_DMSCORE:DMSCORE:BWA_MEM (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/67/c2a06b1c7b4781da339f3e7aacd574] +May-07 23:02:17.585 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-07 23:02:17.586 [Task submitter] INFO nextflow.Session - [48/c69654] Submitted process > NFCORE_DMSCORE:DMSCORE:FASTQC (gid1a_1_quality_1_pe) +May-07 23:02:17.748 [TaskFinalizer-4] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<1:1> +May-07 23:02:17.749 [TaskFinalizer-4] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<2:1> +May-07 23:02:17.749 [TaskFinalizer-4] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:BWA_MEM > Skipping output binding because one or more optional files are missing: fileoutparam<3:1> +May-07 23:02:17.828 [Actor Thread 14] WARN nextflow.processor.TaskContext - Cannot serialize context map. Cause: java.lang.IllegalArgumentException: Unable to create serializer "com.esotericsoftware.kryo.serializers.FieldSerializer" for class: java.lang.ref.ReferenceQueue -- Resume will not work on this process +May-07 23:02:28.533 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 4; name: NFCORE_DMSCORE:DMSCORE:FASTQC (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/48/c69654e9b29a194ea81b7c28f63819] +May-07 23:02:28.582 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-07 23:02:28.583 [Task submitter] INFO nextflow.Session - [4e/7ee9bd] Submitted process > NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS (gid1a_1_quality_1_pe) +May-07 23:02:28.752 [Actor Thread 12] DEBUG nextflow.sort.BigSort - Sort completed -- entries: 2; slices: 1; internal sort time: 0.013 s; external sort time: 0.008 s; total time: 0.021 s +May-07 23:02:28.776 [Actor Thread 12] DEBUG nextflow.file.FileCollector - Saved collect-files list to: /Users/benjaminwehnert/dmscore/work/collect-file/c1149f4a215bbe0651400d7f32cdafdc +May-07 23:02:28.784 [Actor Thread 12] DEBUG nextflow.file.FileCollector - Deleting file collector temp dir: /var/folders/r0/ldrzd4wn1s3516hy0vsn8xzm0000gn/T/nxf-15345989576056151242 +May-07 23:02:32.866 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 6; name: NFCORE_DMSCORE:DMSCORE:BAMFILTER_DMS (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/4e/7ee9bd66291d03ed6f3c2bbdfefe0d] +May-07 23:02:32.872 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-07 23:02:32.873 [Task submitter] INFO nextflow.Session - [55/5dd043] Submitted process > NFCORE_DMSCORE:DMSCORE:MULTIQC +May-07 23:02:43.262 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 7; name: NFCORE_DMSCORE:DMSCORE:MULTIQC; status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/55/5dd0430083da5c332e7950544ac6e0] +May-07 23:02:43.292 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-07 23:02:43.294 [Task submitter] INFO nextflow.Session - [aa/6e83ae] Submitted process > NFCORE_DMSCORE:DMSCORE:PREMERGE (gid1a_1_quality_1_pe) +May-07 23:02:43.298 [TaskFinalizer-7] DEBUG nextflow.processor.TaskProcessor - Process NFCORE_DMSCORE:DMSCORE:MULTIQC > Skipping output binding because one or more optional files are missing: fileoutparam<2> +May-07 23:02:46.384 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 8; name: NFCORE_DMSCORE:DMSCORE:PREMERGE (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/aa/6e83aebc1b031afabb4d8df09e97f3] +May-07 23:02:46.488 [Task submitter] DEBUG n.executor.local.LocalTaskHandler - Launch cmd line: /bin/bash -ue .command.run +May-07 23:02:46.489 [Task submitter] INFO nextflow.Session - [f2/f878d3] Submitted process > NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe) +May-07 23:03:05.753 [Task monitor] DEBUG n.processor.TaskPollingMonitor - Task completed > TaskHandler[id: 9; name: NFCORE_DMSCORE:DMSCORE:GATK_SATURATIONMUTAGENESIS (gid1a_1_quality_1_pe); status: COMPLETED; exit: 0; error: -; workDir: /Users/benjaminwehnert/dmscore/work/f2/f878d301d101b20d245c27be19bbf3] +May-07 23:03:05.837 [Actor Thread 2] DEBUG nextflow.processor.TaskProcessor - Handling unexpected condition for + task: name=NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_PROCESS_GATK (1); work-dir=null + error [nextflow.exception.ProcessUnrecoverableException]: Path value cannot be null +May-07 23:03:05.858 [Actor Thread 2] ERROR nextflow.processor.TaskProcessor - Error executing process > 'NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_PROCESS_GATK (1)' + +Caused by: + Path value cannot be null + -1 error - at org.codehaus.groovy.control.ErrorCollector.failIfErrors(ErrorCollector.java:292) - at org.codehaus.groovy.control.ErrorCollector.addFatalError(ErrorCollector.java:148) - at org.apache.groovy.parser.antlr4.AstBuilder.collectSyntaxError(AstBuilder.java:4753) - at org.apache.groovy.parser.antlr4.AstBuilder.access$100(AstBuilder.java:169) - at org.apache.groovy.parser.antlr4.AstBuilder$3.syntaxError(AstBuilder.java:4764) - at groovyjarjarantlr4.v4.runtime.ProxyErrorListener.syntaxError(ProxyErrorListener.java:44) - at groovyjarjarantlr4.v4.runtime.Parser.notifyErrorListeners(Parser.java:543) - at groovyjarjarantlr4.v4.runtime.DefaultErrorStrategy.notifyErrorListeners(DefaultErrorStrategy.java:154) - at org.apache.groovy.parser.antlr4.internal.DescriptiveErrorStrategy.reportInputMismatch(DescriptiveErrorStrategy.java:104) - at org.apache.groovy.parser.antlr4.internal.DescriptiveErrorStrategy.recover(DescriptiveErrorStrategy.java:55) - at org.apache.groovy.parser.antlr4.internal.DescriptiveErrorStrategy.recoverInline(DescriptiveErrorStrategy.java:68) - at groovyjarjarantlr4.v4.runtime.Parser.match(Parser.java:213) - at org.apache.groovy.parser.antlr4.GroovyParser.compilationUnit(GroovyParser.java:368) - at org.apache.groovy.parser.antlr4.AstBuilder.buildCST(AstBuilder.java:243) - at org.apache.groovy.parser.antlr4.AstBuilder.buildCST(AstBuilder.java:221) - at org.apache.groovy.parser.antlr4.AstBuilder.buildAST(AstBuilder.java:262) - at org.apache.groovy.parser.antlr4.Antlr4ParserPlugin.buildAST(Antlr4ParserPlugin.java:58) - at org.codehaus.groovy.control.SourceUnit.buildAST(SourceUnit.java:256) - at java.base/java.util.Iterator.forEachRemaining(Iterator.java:133) - at java.base/java.util.Spliterators$IteratorSpliterator.forEachRemaining(Spliterators.java:1845) - at java.base/java.util.stream.ReferencePipeline$Head.forEach(ReferencePipeline.java:762) - at org.codehaus.groovy.control.CompilationUnit.compile(CompilationUnit.java:663) - at ch.artecat.grengine.code.groovy.DefaultGroovyCompiler.compile(DefaultGroovyCompiler.java:229) - ... 63 common frames omitted +Tip: you can replicate the issue by changing to the process work dir and entering the command `bash .command.run` +May-07 23:03:05.860 [Actor Thread 2] INFO nextflow.Session - Execution cancelled -- Finishing pending tasks before exit +May-07 23:03:05.865 [Actor Thread 2] ERROR nextflow.Nextflow - Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting +May-07 23:03:05.866 [main] DEBUG nextflow.Session - Session await > all processes finished +May-07 23:03:05.867 [Task monitor] DEBUG n.processor.TaskPollingMonitor - <<< barrier arrives (monitor: local) - terminating tasks monitor poll loop +May-07 23:03:05.867 [main] DEBUG nextflow.Session - Session await > all barriers passed +May-07 23:03:05.868 [Actor Thread 12] DEBUG nextflow.processor.TaskProcessor - Handling unexpected condition for + task: name=NFCORE_DMSCORE:DMSCORE:DMSANALYSIS_PROCESS_GATK; work-dir=null + error [java.lang.InterruptedException]: java.lang.InterruptedException +May-07 23:03:05.872 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'TaskFinalizer' shutdown completed (hard=false) +May-07 23:03:05.872 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'PublishDir' shutdown completed (hard=false) +May-07 23:03:05.880 [main] INFO nextflow.Nextflow - -[nf-core/dmscore] Pipeline completed with errors- +May-07 23:03:05.897 [main] DEBUG n.trace.WorkflowStatsObserver - Workflow completed > WorkflowStats[succeededCount=9; failedCount=0; ignoredCount=0; cachedCount=0; pendingCount=0; submittedCount=0; runningCount=0; retriesCount=0; abortedCount=0; succeedDuration=2m 24s; failedDuration=0ms; cachedDuration=0ms;loadCpus=0; loadMemory=0; peakRunning=2; peakCpus=8; peakMemory=16 GB; ] +May-07 23:03:05.898 [main] DEBUG nextflow.trace.TraceFileObserver - Workflow completed -- saving trace file +May-07 23:03:05.900 [main] DEBUG nextflow.trace.ReportObserver - Workflow completed -- rendering execution report +May-07 23:03:07.314 [main] DEBUG nextflow.trace.TimelineObserver - Workflow completed -- rendering execution timeline +May-07 23:03:07.511 [main] DEBUG nextflow.cache.CacheDB - Closing CacheDB done +May-07 23:03:07.545 [main] INFO org.pf4j.AbstractPluginManager - Stop plugin 'nf-schema@2.3.0' +May-07 23:03:07.545 [main] DEBUG nextflow.plugin.BasePlugin - Plugin stopped nf-schema +May-07 23:03:07.546 [main] DEBUG nextflow.util.ThreadPoolManager - Thread pool 'FileTransfer' shutdown completed (hard=false) +May-07 23:03:07.547 [main] DEBUG nextflow.script.ScriptRunner - > Execution complete -- Goodbye diff --git a/modules/local/gatk/gatktodimsum.nf b/modules/local/gatk/gatktodimsum.nf new file mode 100644 index 0000000..7394663 --- /dev/null +++ b/modules/local/gatk/gatktodimsum.nf @@ -0,0 +1,91 @@ +// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) +// https://github.com/nf-core/modules/tree/master/modules/nf-core/ +// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: +// https://nf-co.re/join +// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. +// All other parameters MUST be provided using the "task.ext" directive, see here: +// https://www.nextflow.io/docs/latest/process.html#ext +// where "task.ext" is a string. +// Any parameters that need to be evaluated in the context of a particular sample +// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. +// TODO nf-core: Software that can be piped together SHOULD be added to separate module files +// unless there is a run-time, storage advantage in implementing in this way +// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: +// bwa mem | samtools view -B -T ref.fasta +// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty +// list (`[]`) instead of a file can be used to work around this issue. + +process GATK_GATKTODIMSUM { + tag "$meta.id" + label 'process_single' + + // TODO nf-core: List required Conda package(s). + // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). + // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. + // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/gatk:3.8--py27_1': + 'biocontainers/gatk:3.8--py27_1' }" + + input: + // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" + // MUST be provided as an input via a Groovy Map called "meta". + // This information may not be required in some instances e.g. indexing reference genome files: + // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf + // TODO nf-core: Where applicable please provide/convert compressed files as input/output + // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. + tuple val(meta), path(bam) + + output: + // TODO nf-core: Named file extensions MUST be emitted for ALL output channels + tuple val(meta), path("*.bam"), emit: bam + // TODO nf-core: List additional required output channels/values here + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 + // If the software is unable to output a version number on the command-line then it can be manually specified + // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf + // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) + // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive + // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter + // using the Nextflow "task" variable e.g. "--threads $task.cpus" + // TODO nf-core: Please replace the example samtools command below with your module's command + // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) + """ + samtools \\ + sort \\ + $args \\ + -@ $task.cpus \\ + -o ${prefix}.bam \\ + -T $prefix \\ + $bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + gatk: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ + + stub: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + // TODO nf-core: A stub section should mimic the execution of the original module as best as possible + // Have a look at the following examples: + // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 + // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 + """ + touch ${prefix}.bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + gatk: \$(samtools --version |& sed '1!d ; s/samtools //') + END_VERSIONS + """ +} From 6ad8e741ed499125fe3355e65480f12b1513a700 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:49:07 +0200 Subject: [PATCH 185/317] Update gatktodimsum.nf --- modules/local/gatk/gatktodimsum.nf | 83 ++++++------------------------ 1 file changed, 16 insertions(+), 67 deletions(-) diff --git a/modules/local/gatk/gatktodimsum.nf b/modules/local/gatk/gatktodimsum.nf index 7394663..891d45b 100644 --- a/modules/local/gatk/gatktodimsum.nf +++ b/modules/local/gatk/gatktodimsum.nf @@ -1,91 +1,40 @@ -// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) -// https://github.com/nf-core/modules/tree/master/modules/nf-core/ -// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: -// https://nf-co.re/join -// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. -// All other parameters MUST be provided using the "task.ext" directive, see here: -// https://www.nextflow.io/docs/latest/process.html#ext -// where "task.ext" is a string. -// Any parameters that need to be evaluated in the context of a particular sample -// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. -// TODO nf-core: Software that can be piped together SHOULD be added to separate module files -// unless there is a run-time, storage advantage in implementing in this way -// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: -// bwa mem | samtools view -B -T ref.fasta -// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty -// list (`[]`) instead of a file can be used to work around this issue. - process GATK_GATKTODIMSUM { tag "$meta.id" label 'process_single' - // TODO nf-core: List required Conda package(s). - // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). - // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. - // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/gatk:3.8--py27_1': - 'biocontainers/gatk:3.8--py27_1' }" + container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" input: - // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" - // MUST be provided as an input via a Groovy Map called "meta". - // This information may not be required in some instances e.g. indexing reference genome files: - // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf - // TODO nf-core: Where applicable please provide/convert compressed files as input/output - // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. - tuple val(meta), path(bam) + tuple val(meta), path(variantCounts_filtered_by_library) + path wt_seq + val pos_range + path script // R script output: - // TODO nf-core: Named file extensions MUST be emitted for ALL output channels - tuple val(meta), path("*.bam"), emit: bam - // TODO nf-core: List additional required output channels/values here - path "versions.yml" , emit: versions + tuple val(meta), path("dimsum_input.tsv"), emit: dimsum_input + path "versions.yml", emit: versions when: task.ext.when == null || task.ext.when script: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 - // If the software is unable to output a version number on the command-line then it can be manually specified - // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf - // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) - // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive - // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter - // using the Nextflow "task" variable e.g. "--threads $task.cpus" - // TODO nf-core: Please replace the example samtools command below with your module's command - // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) """ - samtools \\ - sort \\ - $args \\ - -@ $task.cpus \\ - -o ${prefix}.bam \\ - -T $prefix \\ - $bam + start_stop_codon="$pos_range" + + Rscript -e "source('$script'); generate_dimsum_input('$wt_seq', '$variantCounts_filtered_by_library', '\$start_stop_codon', 'dimsum_input.tsv')" + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') cat <<-END_VERSIONS > versions.yml - "${task.process}": - gatk: \$(samtools --version |& sed '1!d ; s/samtools //') + GATK_GATKTODIMSUM: + r-base: \$R_VERSION END_VERSIONS """ stub: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: A stub section should mimic the execution of the original module as best as possible - // Have a look at the following examples: - // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 - // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 """ - touch ${prefix}.bam - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - gatk: \$(samtools --version |& sed '1!d ; s/samtools //') - END_VERSIONS + touch dimsum_input.tsv + echo "GATK_GATKTODIMSUM:" > versions.yml + echo " stub-version: 0.0.0" >> versions.yml """ } From 1156f2d7fd74742deb07870df78af0810327c593 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 11:51:26 +0200 Subject: [PATCH 186/317] Update dmscore.nf --- workflows/dmscore.nf | 9 +++++++++ 1 file changed, 9 insertions(+) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 801152b..aa8bb89 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -19,6 +19,7 @@ include { VISUALIZATION_GLOBAL_POS_BIASES_COUNTS } from '../modules/local/v include { VISUALIZATION_GLOBAL_POS_BIASES_COV } from '../modules/local/visualization/visualization' include { VISUALIZATION_LOGDIFF } from '../modules/local/visualization/visualization' include { VISUALIZATION_SEQDEPTH } from '../modules/local/visualization/visualization' +include { GATK_GATKTODIMSUM } from '../modules/local/gatk/saturationmutagenesis' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -278,6 +279,14 @@ workflow DMSCORE { ) } + GATK_GATKTODIMSUM( + variantCounts_filtered_by_library_ch, + ch_fasta, + reading_frame_ch, + gatk_to_dimsum_script_ch + ) + } + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From 51e8e91f79886b41963b588441901b33541987e5 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 12:31:13 +0200 Subject: [PATCH 187/317] Update modules.config --- conf/modules.config | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index eb49bec..7bb7896 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -143,4 +143,12 @@ process { saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } + + withName: 'GATK_GATKTODIMSUM' { + publishDir = [ + path: "${params.outdir}/DiMSum", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } } From 51f497e85ea150e3408dc974180644094db0afc2 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 12:32:27 +0200 Subject: [PATCH 188/317] Update dmscore.nf --- workflows/dmscore.nf | 11 +++++------ 1 file changed, 5 insertions(+), 6 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index aa8bb89..4624fb2 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -280,12 +280,11 @@ workflow DMSCORE { } GATK_GATKTODIMSUM( - variantCounts_filtered_by_library_ch, - ch_fasta, - reading_frame_ch, - gatk_to_dimsum_script_ch - ) - } + variantCounts_filtered_by_library_ch, + ch_fasta, + reading_frame_ch, + gatk_to_dimsum_script_ch + ) emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html From 930e9b4f7eb3ccd466110ad4ae3e826609be24da Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 12:33:23 +0200 Subject: [PATCH 189/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 4624fb2..2996646 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -19,7 +19,7 @@ include { VISUALIZATION_GLOBAL_POS_BIASES_COUNTS } from '../modules/local/v include { VISUALIZATION_GLOBAL_POS_BIASES_COV } from '../modules/local/visualization/visualization' include { VISUALIZATION_LOGDIFF } from '../modules/local/visualization/visualization' include { VISUALIZATION_SEQDEPTH } from '../modules/local/visualization/visualization' -include { GATK_GATKTODIMSUM } from '../modules/local/gatk/saturationmutagenesis' +include { GATK_GATKTODIMSUM } from '../modules/local/gatk/gatktodimsum' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' From 23e325b024f7d1a93d20a65dc929318c66f5d634 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 12:46:47 +0200 Subject: [PATCH 190/317] Update dmscore.nf From fdb6365c21b6b4eac836452d165e6011e16d85de Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 12:48:18 +0200 Subject: [PATCH 191/317] Update dmscore.nf --- workflows/dmscore.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 2996646..265de6d 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -281,7 +281,7 @@ workflow DMSCORE { GATK_GATKTODIMSUM( variantCounts_filtered_by_library_ch, - ch_fasta, + ch_fasta.map{ it[1] }, reading_frame_ch, gatk_to_dimsum_script_ch ) From 78402d05624607a7fd3cbf332c9e78cb02677c52 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 12:53:40 +0200 Subject: [PATCH 192/317] Update nextflow_schema.json --- nextflow_schema.json | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/nextflow_schema.json b/nextflow_schema.json index cc08d2f..1c4eaf0 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -63,6 +63,11 @@ "format": "file-path", "description": "Path to a comma-separated .txt file listing custom codons (e.g., 'AAA,AAC,AAG,...'). Required when mutagenesis_type is 'custom'." } + "run_seqdepth": { + "type": "boolean", + "default": false, + "description": "Whether to run the SeqDepth simulation module. This is computationally intensive and optional." + } } }, "reference_genome_options": { From 45174d54170f3a4d56ce6f62961ded354de91cee Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Fri, 9 May 2025 12:54:33 +0200 Subject: [PATCH 193/317] Update nextflow_schema.json --- nextflow_schema.json | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 1c4eaf0..3db9ee6 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -62,12 +62,12 @@ "type": "string", "format": "file-path", "description": "Path to a comma-separated .txt file listing custom codons (e.g., 'AAA,AAC,AAG,...'). Required when mutagenesis_type is 'custom'." - } + }, "run_seqdepth": { "type": "boolean", "default": false, "description": "Whether to run the SeqDepth simulation module. This is computationally intensive and optional." - } + } } }, "reference_genome_options": { From 8e137853682dd01d00ffca40766cab83f6ea8999 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 2 Jul 2025 12:55:16 +0200 Subject: [PATCH 194/317] Update nextflow.config to implement local memory and CPU requirements --- nextflow.config | 34 ++++++++++++++++++++++++++++++++++ 1 file changed, 34 insertions(+) diff --git a/nextflow.config b/nextflow.config index 256e8ea..59f979b 100644 --- a/nextflow.config +++ b/nextflow.config @@ -160,6 +160,40 @@ profiles { ] } } + local { + process { + withLabel:process_single { + cpus = 4 + memory = 8.GB + time = '6h' + } + withLabel:process_low { + cpus = 4 + memory = 8.GB + time = '6h' + } + withLabel:process_medium { + cpus = 4 + memory = 8.GB + time = '6h' + } + withLabel:process_high { + cpus = 4 + memory = 8.GB + time = '6h' + } + withLabel:process_long { + cpus = 4 + memory = 8.GB + time = '6h' + } + withLabel:process_high_memory { + cpus = 4 + memory = 8.GB + time = '6h' + } + } + } test { includeConfig 'conf/test.config' } test_full { includeConfig 'conf/test_full.config' } } From c47b3bb2c9005227297b40f3eb32d456a31532c7 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 2 Jul 2025 13:18:10 +0200 Subject: [PATCH 195/317] Update nextflow.config to add slurm config --- nextflow.config | 11 ++++++++++- 1 file changed, 10 insertions(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index 59f979b..cd444c1 100644 --- a/nextflow.config +++ b/nextflow.config @@ -193,7 +193,16 @@ profiles { time = '6h' } } - } + } + slurm { + // Activate executor and Wave + process.executor = 'slurm' + wave.enabled = true + wave.strategy = 'container' + singularity.enabled = true + singularity.autoMounts = true + singularity.cacheDir = "$HOME/.singularity" + } test { includeConfig 'conf/test.config' } test_full { includeConfig 'conf/test_full.config' } } From a031260bc5f1a8e4d58967894c5ef842d032401d Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 2 Jul 2025 13:33:25 +0200 Subject: [PATCH 196/317] Update nextflow.config --- nextflow.config | 3 +++ 1 file changed, 3 insertions(+) diff --git a/nextflow.config b/nextflow.config index cd444c1..f334d9e 100644 --- a/nextflow.config +++ b/nextflow.config @@ -202,6 +202,9 @@ profiles { singularity.enabled = true singularity.autoMounts = true singularity.cacheDir = "$HOME/.singularity" + env { + NXF_SINGULARITY_PULL_DOCKER_CONTAINER = true // allows to pull docker as singularity image + } } test { includeConfig 'conf/test.config' } test_full { includeConfig 'conf/test_full.config' } From 8b75011a39b1e1c0629baa75b4a536eb8d8a2709 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 2 Jul 2025 13:36:21 +0200 Subject: [PATCH 197/317] Update nextflow.config --- nextflow.config | 2 -- 1 file changed, 2 deletions(-) diff --git a/nextflow.config b/nextflow.config index f334d9e..f7fd4cc 100644 --- a/nextflow.config +++ b/nextflow.config @@ -197,8 +197,6 @@ profiles { slurm { // Activate executor and Wave process.executor = 'slurm' - wave.enabled = true - wave.strategy = 'container' singularity.enabled = true singularity.autoMounts = true singularity.cacheDir = "$HOME/.singularity" From 30080a1f36b99e6c8fa7993faaf570d9b9998cd4 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Wed, 2 Jul 2025 14:57:08 +0200 Subject: [PATCH 198/317] Update nextflow.config --- nextflow.config | 10 ---------- 1 file changed, 10 deletions(-) diff --git a/nextflow.config b/nextflow.config index f7fd4cc..585ddad 100644 --- a/nextflow.config +++ b/nextflow.config @@ -194,16 +194,6 @@ profiles { } } } - slurm { - // Activate executor and Wave - process.executor = 'slurm' - singularity.enabled = true - singularity.autoMounts = true - singularity.cacheDir = "$HOME/.singularity" - env { - NXF_SINGULARITY_PULL_DOCKER_CONTAINER = true // allows to pull docker as singularity image - } - } test { includeConfig 'conf/test.config' } test_full { includeConfig 'conf/test_full.config' } } From c42043dad5f61d45a4795578697fe6bbb729a588 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Mon, 21 Jul 2025 16:07:38 +0200 Subject: [PATCH 199/317] Update possiblemutations.nf --- modules/local/dmsanalysis/possiblemutations.nf | 5 ++++- 1 file changed, 4 insertions(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/possiblemutations.nf b/modules/local/dmsanalysis/possiblemutations.nf index ea7d9cf..344eb0b 100644 --- a/modules/local/dmsanalysis/possiblemutations.nf +++ b/modules/local/dmsanalysis/possiblemutations.nf @@ -3,7 +3,10 @@ process DMSANALYSIS_POSSIBLE_MUTATIONS { label 'process_single' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" input: tuple val(meta), path(wt_seq) From 6ce8e7fb181e570692500fa0747862740206c10f Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Mon, 21 Jul 2025 16:18:23 +0200 Subject: [PATCH 200/317] Update aaseq.nf --- modules/local/dmsanalysis/aaseq.nf | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/aaseq.nf b/modules/local/dmsanalysis/aaseq.nf index ec45ec1..8d02563 100644 --- a/modules/local/dmsanalysis/aaseq.nf +++ b/modules/local/dmsanalysis/aaseq.nf @@ -3,7 +3,9 @@ process DMSANALYSIS_AASEQ { label 'process_single' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" input: tuple val(meta), path(wt_seq) From a78e3ddc7c4e2e97fdd905f1a975ab808e797201 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Mon, 21 Jul 2025 16:42:37 +0200 Subject: [PATCH 201/317] Update processgatk.nf --- modules/local/dmsanalysis/processgatk.nf | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/processgatk.nf b/modules/local/dmsanalysis/processgatk.nf index 6c60f1d..50bc4e0 100644 --- a/modules/local/dmsanalysis/processgatk.nf +++ b/modules/local/dmsanalysis/processgatk.nf @@ -3,7 +3,9 @@ process DMSANALYSIS_PROCESS_GATK { label 'process_single' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" publishDir "${params.outdir}/intermediate_files", mode: 'copy' From b7c4f11cc1b6d5e8317c47acea7a781a482eccc2 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Mon, 21 Jul 2025 16:43:20 +0200 Subject: [PATCH 202/317] Update gatktodimsum.nf --- modules/local/gatk/gatktodimsum.nf | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/modules/local/gatk/gatktodimsum.nf b/modules/local/gatk/gatktodimsum.nf index 891d45b..9c51a88 100644 --- a/modules/local/gatk/gatktodimsum.nf +++ b/modules/local/gatk/gatktodimsum.nf @@ -3,7 +3,9 @@ process GATK_GATKTODIMSUM { label 'process_single' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" input: tuple val(meta), path(variantCounts_filtered_by_library) From 289786a8ebbd0dacc914ca253bf6fd7d9124926b Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Mon, 21 Jul 2025 16:43:40 +0200 Subject: [PATCH 203/317] Update visualization.nf --- modules/local/visualization/visualization.nf | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/modules/local/visualization/visualization.nf b/modules/local/visualization/visualization.nf index 5bf6cd2..cf8fc2a 100644 --- a/modules/local/visualization/visualization.nf +++ b/modules/local/visualization/visualization.nf @@ -3,7 +3,9 @@ process VISUALIZATION_COUNTS_PER_COV { label 'process_single' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" input: tuple val(meta), path(variantCounts_for_heatmaps) From b51e5226777a7be5a2c8b6df6b51de49d557466d Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Mon, 21 Jul 2025 16:47:47 +0200 Subject: [PATCH 204/317] Update visualization.nf --- modules/local/visualization/visualization.nf | 20 +++++++++++++++----- 1 file changed, 15 insertions(+), 5 deletions(-) diff --git a/modules/local/visualization/visualization.nf b/modules/local/visualization/visualization.nf index cf8fc2a..e997fad 100644 --- a/modules/local/visualization/visualization.nf +++ b/modules/local/visualization/visualization.nf @@ -43,7 +43,9 @@ process VISUALIZATION_COUNTS_HEATMAP { label 'process_single' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" input: tuple val(meta), path(variantCounts_for_heatmaps) @@ -81,7 +83,9 @@ process VISUALIZATION_GLOBAL_POS_BIASES_COUNTS { label 'process_single' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" input: tuple val(meta), path(variantCounts_filtered_by_library) @@ -122,7 +126,9 @@ process VISUALIZATION_GLOBAL_POS_BIASES_COV { label 'process_single' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" input: tuple val(meta), path(variantCounts_filtered_by_library) @@ -162,7 +168,9 @@ process VISUALIZATION_LOGDIFF { label 'process_single' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" input: tuple val(meta), path(library_completed_variantCounts) @@ -201,7 +209,9 @@ process VISUALIZATION_SEQDEPTH { label 'process_high' conda "${moduleDir}/environment.yml" - container "community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" input: tuple val(meta), path(variantCounts_filtered_by_library) From f691dd4db5567ae0c29f2d645c7af850eb44fbd8 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 28 Jul 2025 23:40:55 +0100 Subject: [PATCH 205/317] Update README.md --- README.md | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/README.md b/README.md index b6ae647..6240054 100644 --- a/README.md +++ b/README.md @@ -5,8 +5,7 @@ -[![GitHub Actions CI Status](https://github.com/nf-core/dmscore/actions/workflows/ci.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/ci.yml) -[![GitHub Actions Linting Status](https://github.com/nf-core/dmscore/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/dmscore/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) +(https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/dmscore/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) [![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com) [![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A524.04.2-23aa62.svg)](https://www.nextflow.io/) From 10559646e5f45570f9806754696cfe541c197c2e Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 28 Jul 2025 23:42:08 +0100 Subject: [PATCH 206/317] Update README.md --- README.md | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/README.md b/README.md index 6240054..ecbd195 100644 --- a/README.md +++ b/README.md @@ -5,9 +5,11 @@ + +[![GitHub Actions CI Status](https://github.com/nf-core/dmscore/actions/workflows/ci.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/ci.yml) +[![GitHub Actions Linting Status](https://github.com/nf-core/dmscore/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/linting.yml)[![AWS CI] (https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/dmscore/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) [![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com) - [![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A524.04.2-23aa62.svg)](https://www.nextflow.io/) [![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/) [![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/) From 669b6962ea80895701611033270baacbff77c60b Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 28 Jul 2025 23:42:40 +0100 Subject: [PATCH 207/317] Update README.md --- README.md | 7 +++---- 1 file changed, 3 insertions(+), 4 deletions(-) diff --git a/README.md b/README.md index ecbd195..1311730 100644 --- a/README.md +++ b/README.md @@ -6,18 +6,17 @@ + [![GitHub Actions CI Status](https://github.com/nf-core/dmscore/actions/workflows/ci.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/ci.yml) -[![GitHub Actions Linting Status](https://github.com/nf-core/dmscore/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/linting.yml)[![AWS CI] -(https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/dmscore/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) +[![GitHub Actions Linting Status](https://github.com/nf-core/dmscore/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/dmscore/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) [![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com) + [![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A524.04.2-23aa62.svg)](https://www.nextflow.io/) [![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/) [![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/) [![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/) [![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/dmscore) -[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23dmscore-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/dmscore)[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core) - ## Introduction **nf-core/dmscore** is a bioinformatics pipeline that ... From 432e664f63d034d67d87053770f072b0d96b46f2 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 28 Jul 2025 23:45:04 +0100 Subject: [PATCH 208/317] Update README.md --- README.md | 7 +++---- 1 file changed, 3 insertions(+), 4 deletions(-) diff --git a/README.md b/README.md index 1311730..2febb44 100644 --- a/README.md +++ b/README.md @@ -5,8 +5,6 @@ - - [![GitHub Actions CI Status](https://github.com/nf-core/dmscore/actions/workflows/ci.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/ci.yml) [![GitHub Actions Linting Status](https://github.com/nf-core/dmscore/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/dmscore/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) [![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com) @@ -17,9 +15,10 @@ [![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/) [![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/dmscore) -## Introduction +Overview +**nf-core/dms** is a reproducible, scalable, and community-curated pipeline for analyzing deep mutational scanning (DMS) data using shotgun DNA sequencing. DMS enables researchers to measure the fitness effects of thousands of gene variants simultaneously, helping to classify disease causing mutants in human and animal populations, to learn fundamental rules of virus evolution, protein architecture, splicing or small-molecule interactions. -**nf-core/dmscore** is a bioinformatics pipeline that ... +This pipeline simplifies and standardizes the complex steps involved in shotgun sequencing-based DMS experiments—particularly for long open reading frames (ORFs)—from read alignment to fitness inference and QC reporting. - - -1. Read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/))2. Present QC for raw reads ([`MultiQC`](http://multiqc.info/)) +- Paths to paired-end `.fastq.gz` files from shotgun sequencing +- Their classification as either input or output samples +- Replicate IDs +- Associated experimental metadata -## Usage +See [sample CSV](link) for formatting. -> [!NOTE] -> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/usage/installation) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/usage/introduction#how-to-run-a-pipeline) with `-profile test` before running the workflow on actual data. - - +## Citation -Now, you can run the pipeline using: +If you use this pipeline in your research, please cite: +> _bioRxiv preprint, 2025_ (link coming soon) - +Please also cite the nf-core framework: +> Ewels et al., _Nature Biotechnology_, 2020 +> [https://doi.org/10.1038/s41587-020-0439-x](https://doi.org/10.1038/s41587-020-0439-x) -```bash -nextflow run nf-core/dmscore \ - -profile \ - --input samplesheet.csv \ - --outdir -``` +--- -> [!WARNING] -> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/usage/getting_started/configuration#custom-configuration-files). For more details and further functionality, please refer to the [usage documentation](https://nf-co.re/dmscore/usage) and the [parameter documentation](https://nf-co.re/dmscore/parameters). From 9c5b4a6e028b0f25d707cff3995abe6398d7da9a Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Tue, 29 Jul 2025 00:33:32 +0100 Subject: [PATCH 222/317] Update README.md --- README.md | 58 ++++++++++++++++++++++++------------------------------- 1 file changed, 25 insertions(+), 33 deletions(-) diff --git a/README.md b/README.md index 45489e5..44a3045 100644 --- a/README.md +++ b/README.md @@ -22,7 +22,7 @@ While DNA synthesis and sequencing technologies have advanced substantially, long open reading frame (ORF) targets still present major challenges for DMS studies. Shotgun DNA sequencing can be used to greatly speed up the inference of long ORF mutant fitness landscapes, theoretically at no expense in accuracy. We have designed the **nf-core/dms** pipeline to unlock the power of shotgun sequencing based DMS studies, to simplify and standardise the complex bioinformatics steps involved in data processing of such experiments – from read alignment to QC reporting and fitness landscape inferences. -> 📄 Reference: Wehnert et al., 2025 (bioRxiv preprint, coming soon) +> 📄 Reference: Wehnert et al., _bioRxiv_ preprint (coming soon) --- @@ -60,9 +60,6 @@ Prepare: - A **sample sheet** CSV to specify input/output labels, replicates, etc. (see [example](link)) - A **reference FASTA** file for the gene or region of interest -> [!WARNING] -> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration; see [docs](https://nf-co.re/docs/usage/getting_started/configuration#custom-configuration-files). - To execute **nf-core/dms**, run the basic command: ```bash @@ -127,53 +124,48 @@ results/ --- -## Citation +## 7. Citation If you use this pipeline in your research, please cite: -> _bioRxiv preprint, 2025_ (link coming soon) +> 📄 Reference: Wehnert et al., _bioRxiv_ preprint (coming soon) Please also cite the nf-core framework: -> Ewels et al., _Nature Biotechnology_, 2020 +> 📄 Ewels et al., _Nature Biotechnology_, 2020 > [https://doi.org/10.1038/s41587-020-0439-x](https://doi.org/10.1038/s41587-020-0439-x) --- +## 8. License -For more details and further functionality, please refer to the [usage documentation](https://nf-co.re/dmscore/usage) and the [parameter documentation](https://nf-co.re/dmscore/parameters). - -## Pipeline output - -To see the results of an example test run with a full size dataset refer to the [results](https://nf-co.re/dmscore/results) tab on the nf-core website pipeline page. -For more details about the output files and reports, please refer to the -[output documentation](https://nf-co.re/dmscore/output). - -## Credits +[MIT License](link) -nf-core/dmscore was originally written by Benjamin Wehnert & Max Stammnitz. +© 2025 Benjamin Wehnert, Taylor Mighell, Fei Sang, Ben Lehner, Maximilian Stammnitz -We thank the following people for their extensive assistance in the development of this pipeline: +--- - +## 9. Contributing -## Contributions and Support +We welcome contributions from the community! -If you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md). +- 🛠 Suggest or implement new modules +- 🐛 Report bugs or issues +- ✍️ Help improve the documentation +- 🔁 Request support for new fitness estimation tools -For further information or help, don't hesitate to get in touch on the [Slack `#dmscore` channel](https://nfcore.slack.com/channels/dmscore) (you can join with [this invite](https://nf-co.re/join/slack)). +Please open an [issue](link) or [pull request](link) via this GitHub page. -## Citations +You can also reach out on via **nf-core Slack** in the `#dms` channel ([join here](link)). - - +--- - +## 10. Contact -An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file. +For detailled scientific or technical questions, feel free to reach out to us directly: -You can cite the `nf-core` publication as follows: +- Benjamin Wehnert — wehnertbenjamin@gmail.com +- Taylor Mighell — taylor.mighell@crg.eu +- Fei Sang — fs18@sanger.ac.uk +- Ben Lehner — bl11@sanger.ac.uk +- Maximilian Stammnitz — maximilian.stammnitz@crg.eu -> **The nf-core framework for community-curated bioinformatics pipelines.** -> -> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen. -> -> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x). +--- From de38f6d936c8dbaad4cbdba372fb2db5da456e0a Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Tue, 29 Jul 2025 00:38:42 +0100 Subject: [PATCH 223/317] Update README.md --- README.md | 12 +++++------- 1 file changed, 5 insertions(+), 7 deletions(-) diff --git a/README.md b/README.md index 44a3045..193a810 100644 --- a/README.md +++ b/README.md @@ -147,14 +147,12 @@ Please also cite the nf-core framework: We welcome contributions from the community! -- 🛠 Suggest or implement new modules -- 🐛 Report bugs or issues -- ✍️ Help improve the documentation -- 🔁 Request support for new fitness estimation tools +Please open an [issue](link) or [pull request](link) via this GitHub page, to: +- Suggest or help implementing new modules for custom workflows +- Report bugs and other challenges in running **nf-core/dms** +- Help improve this documentation -Please open an [issue](link) or [pull request](link) via this GitHub page. - -You can also reach out on via **nf-core Slack** in the `#dms` channel ([join here](link)). +You can also reach out to us via the **nf-core Slack** in the `#dms` channel ([join here](link)). --- From 12d2f4b8f0fea4a39d009ea3bcc3aaba7fb8bd8b Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Tue, 29 Jul 2025 00:42:08 +0100 Subject: [PATCH 224/317] Update README.md --- README.md | 14 +++++++------- 1 file changed, 7 insertions(+), 7 deletions(-) diff --git a/README.md b/README.md index 193a810..6689842 100644 --- a/README.md +++ b/README.md @@ -27,12 +27,12 @@ While DNA synthesis and sequencing technologies have advanced substantially, lon --- ## 2. Features of nf-core/dms -- End-to-end analysis of DMS shotgun sequencing data -- Modular, three-stage workflow: alignment → QC → fitness estimation +- End-to-end analyses of DMS shotgun sequencing data +- Modular, three-stage workflow: alignment → QC → error-aware fitness estimation - Integrates with popular statistical tools like [DiMSum](https://github.com/lehner-lab/DiMSum), [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) -- Supports multiple mutagenesis strategies (e.g. nicking by NNK and NNS codons) -- Containerized via Docker/Singularity -- Scalable across HPC and cloud systems +- Supports multiple mutagenesis strategies, e.g. nicking by NNK and NNS codons +- Containerized via Docker, Singularity and Apptainer +- Scalable across HPC and Cloud systems - Monitors CPU, memory, and CO₂ usage --- @@ -152,13 +152,13 @@ Please open an [issue](link) or [pull request](link) via this GitHub page, to: - Report bugs and other challenges in running **nf-core/dms** - Help improve this documentation -You can also reach out to us via the **nf-core Slack** in the `#dms` channel ([join here](link)). +You can also reach out to us via the **nf-core Slack**, by use of the `#dms` channel ([join here](link)). --- ## 10. Contact -For detailled scientific or technical questions, feel free to reach out to us directly: +For detailled scientific or technical questions, feedback and experimental discussions, feel free to contact us directly: - Benjamin Wehnert — wehnertbenjamin@gmail.com - Taylor Mighell — taylor.mighell@crg.eu From 506ef1c6d51b94fa53fe33c715f570679d3e74fb Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Tue, 29 Jul 2025 00:45:10 +0100 Subject: [PATCH 225/317] Update README.md --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 6689842..977f6c9 100644 --- a/README.md +++ b/README.md @@ -127,7 +127,7 @@ results/ ## 7. Citation If you use this pipeline in your research, please cite: -> 📄 Reference: Wehnert et al., _bioRxiv_ preprint (coming soon) +> 📄 Wehnert et al., _bioRxiv_ preprint (coming soon) Please also cite the nf-core framework: > 📄 Ewels et al., _Nature Biotechnology_, 2020 From 874c7b6a3d84c9cc570f5e69f76f190d981ace0b Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 7 Aug 2025 20:22:15 +0200 Subject: [PATCH 226/317] Update README.md Added functional hyperlinks --- README.md | 9 +++------ 1 file changed, 3 insertions(+), 6 deletions(-) diff --git a/README.md b/README.md index 977f6c9..1ad60a6 100644 --- a/README.md +++ b/README.md @@ -57,7 +57,7 @@ nextflow run nf-core/dms -profile docker ## 4. Usage Prepare: -- A **sample sheet** CSV to specify input/output labels, replicates, etc. (see [example](link)) +- A **sample sheet** CSV to specify input/output labels, replicates, etc. (see [example](assets/samplesheet.csv)) - A **reference FASTA** file for the gene or region of interest To execute **nf-core/dms**, run the basic command: @@ -147,12 +147,12 @@ Please also cite the nf-core framework: We welcome contributions from the community! -Please open an [issue](link) or [pull request](link) via this GitHub page, to: +Please open an [issue](../../issues/new) or [pull request](../../compare) via this GitHub page, to: - Suggest or help implementing new modules for custom workflows - Report bugs and other challenges in running **nf-core/dms** - Help improve this documentation -You can also reach out to us via the **nf-core Slack**, by use of the `#dms` channel ([join here](link)). +You can also reach out to us via the **nf-core Slack**, by use of the `#dms` channel ([join here](https://join.slack.com/share/enQtOTMyMDc3MTA0Mzg0Mi04YmRiNDEwZTBlOTRiN2M2ZGU5ZGVmOWQ3YzA0YjA4NzhiNjFhNTVlNDA4ZTZjOTE2MjE5MmIzYWZjZTljMTE3)). --- @@ -161,9 +161,6 @@ You can also reach out to us via the **nf-core Slack**, by use of the `#dms` cha For detailled scientific or technical questions, feedback and experimental discussions, feel free to contact us directly: - Benjamin Wehnert — wehnertbenjamin@gmail.com -- Taylor Mighell — taylor.mighell@crg.eu -- Fei Sang — fs18@sanger.ac.uk -- Ben Lehner — bl11@sanger.ac.uk - Maximilian Stammnitz — maximilian.stammnitz@crg.eu --- From 524e26e2da250bb5c32cf8144a8beac3605eae34 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 7 Aug 2025 20:26:21 +0200 Subject: [PATCH 227/317] Update README.md Added functional hyperlinks --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 1ad60a6..840e944 100644 --- a/README.md +++ b/README.md @@ -105,7 +105,7 @@ The primary pipeline input is a sample sheet `.csv` file listing: - Replicate IDs - Associated experimental metadata -See [sample CSV](link) for formatting. +See [sample CSV](assets/samplesheet.csv) for formatting. --- From beebdeaea0a6950214573e17547faad78519cb8e Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Fri, 8 Aug 2025 23:56:48 +0200 Subject: [PATCH 228/317] Update README.md --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 840e944..9a868ed 100644 --- a/README.md +++ b/README.md @@ -20,7 +20,7 @@ ## 1. Overview **nf-core/dms** is a reproducible, scalable, and community-curated pipeline for analyzing deep mutational scanning (DMS) data using shotgun DNA sequencing. DMS enables researchers to measure the fitness effects of thousands of gene variants simultaneously, helping to classify disease causing mutants in human and animal populations, to learn fundamental rules of virus evolution, protein architecture, splicing or small-molecule interactions. -While DNA synthesis and sequencing technologies have advanced substantially, long open reading frame (ORF) targets still present major challenges for DMS studies. Shotgun DNA sequencing can be used to greatly speed up the inference of long ORF mutant fitness landscapes, theoretically at no expense in accuracy. We have designed the **nf-core/dms** pipeline to unlock the power of shotgun sequencing based DMS studies, to simplify and standardise the complex bioinformatics steps involved in data processing of such experiments – from read alignment to QC reporting and fitness landscape inferences. +While DNA synthesis and sequencing technologies have advanced substantially, long open reading frame (ORF) targets still present major challenges for DMS studies. Shotgun DNA sequencing can be used to greatly speed up the inference of long ORF mutant fitness landscapes, theoretically at no expense in accuracy. We have designed the **nf-core/dms** pipeline to unlock the power of shotgun sequencing based DMS studies on long ORFs, to simplify and standardise the complex bioinformatics steps involved in data processing of such experiments – from read alignment to QC reporting and fitness landscape inferences. > 📄 Reference: Wehnert et al., _bioRxiv_ preprint (coming soon) From 4db09be47334892b45e1fa074bb0959109a44b96 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Mon, 25 Aug 2025 17:01:29 +0200 Subject: [PATCH 229/317] first part for multiple samples processing --- conf/modules.config | 4 +- .../local/bamprocessing/bamfilteringdms.nf | 2 +- modules/local/bamprocessing/premerge.nf | 4 +- workflows/dmscore.nf | 58 ++++++++++++++----- 4 files changed, 49 insertions(+), 19 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 7bb7896..73b499e 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -50,7 +50,7 @@ process { withName: 'BAMFILTER_DMS' { publishDir = [ - path: "${params.outdir}/intermediate_files/bam_files", + path: "${params.outdir}/intermediate_files/bam_files/filtered", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] @@ -58,7 +58,7 @@ process { withName: 'PREMERGE' { publishDir = [ - path: "${params.outdir}/intermediate_files/bam_files", + path: "${params.outdir}/intermediate_files/bam_files/premerged", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] diff --git a/modules/local/bamprocessing/bamfilteringdms.nf b/modules/local/bamprocessing/bamfilteringdms.nf index 22be831..16603b7 100644 --- a/modules/local/bamprocessing/bamfilteringdms.nf +++ b/modules/local/bamprocessing/bamfilteringdms.nf @@ -64,7 +64,7 @@ process BAMFILTER_DMS { awk '{if(\$6 !~ /I/ && \$6 !~ /D/ && \$6 !~ /N/) print \$0}' | \ samtools view -h | \ awk '{for(i=1;i<=NF;i++) if(\$i ~ /^NM:i:/ && \$i != "NM:i:0") {print \$0; next}} \$1 ~ /^@/' | \ - samtools view -bS > filtered_reads.bam + samtools view -bS > ${meta.id}_filtered.bam cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/local/bamprocessing/premerge.nf b/modules/local/bamprocessing/premerge.nf index b1bb305..8f8b44a 100644 --- a/modules/local/bamprocessing/premerge.nf +++ b/modules/local/bamprocessing/premerge.nf @@ -27,7 +27,7 @@ process PREMERGE { path wt_seq output: - tuple val(meta), path("merged_reads.bam"), emit: bam + tuple val(meta), path("*.bam"), emit: bam path "versions.yml", emit: versions when: @@ -44,7 +44,7 @@ process PREMERGE { # Re-align merged reads bwa index $wt_seq - bwa mem $wt_seq merged_reads.fastq | samtools view -Sb - > merged_reads.bam + bwa mem $wt_seq merged_reads.fastq | samtools view -Sb - > ${prefix}_merged.bam # Save version information cat <<-END_VERSIONS > versions.yml diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 265de6d..68e5447 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -183,29 +183,59 @@ workflow DMSCORE { // Broadcast index to all samples ch_bwa_index = BWA_INDEX.out.index - BWA_MEM ( - ch_samplesheet, // each sample's reads: tuple [meta, reads] - ch_bwa_index, // broadcasted index files - ch_fasta, // broadcasted fasta file - false // sort_bam + + + + // Broadcast the index to all samples + ch_bwa_index_broadcast = ch_samplesheet + .combine(ch_bwa_index) + .map { [it[2], it[3]] } + + // Broadcast the fasta to all samples + ch_fasta_broadcast = ch_fasta + .combine(ch_samplesheet) + .map { [it[0], it[1]] } + + // Broadcast the sort flag to all samples + ch_sort_bam = ch_samplesheet.map { false } + + // Run BWA_MEM with all four inputs aligned + BWA_MEM( + ch_samplesheet, + ch_bwa_index_broadcast, + ch_fasta_broadcast, + ch_sort_bam ) + BAMFILTER_DMS ( BWA_MEM.out.bam ) - PREMERGE ( - BAMFILTER_DMS.out.bam, - ch_fasta.map{ it[1] } // extract fasta path from Tuple - ) + // Broadcast the FASTA path to every BAM emitted by BAMFILTER_DMS + ch_fasta_path_broadcast = ch_fasta + .combine(BAMFILTER_DMS.out.bam) // flattened item: [meta3, fasta, meta, bam] + .map { it[1] } // keep only the fasta path (N emissions) - GATK_SATURATIONMUTAGENESIS ( - PREMERGE.out.bam, // merged reads (tuple(meta, merged_reads.fastq)) - ch_fasta.map{ it[1] }, // fasta path (path) - reading_frame_ch, // codon range (path) - min_counts_ch // min_counts (val) + PREMERGE( + BAMFILTER_DMS.out.bam, // tuple(val(meta), path(bam)) + ch_fasta_path_broadcast // path(fasta) ) +// FASTA path for GATK: broadcast to N +ch_fasta_for_gatk = ch_fasta.combine(PREMERGE.out.bam).map { it[1] } // path -- N +// Reading frame for GATK: broadcast to N (it's a val string) +ch_rf_for_gatk = reading_frame_ch.combine(PREMERGE.out.bam).map { it[0] } // val -- N +// min_counts for GATK: broadcast to N (also a val) +ch_min_for_gatk = min_counts_ch.combine(PREMERGE.out.bam).map { it[0] } // val -- N + +GATK_SATURATIONMUTAGENESIS( + PREMERGE.out.bam, // merged reads - tuple(val(meta), path(bam)) + ch_fasta_for_gatk, // path(fasta) + ch_rf_for_gatk, // val(reading_frame string) + ch_min_for_gatk // val(min_counts) +) + DMSANALYSIS_AASEQ ( ch_fasta, reading_frame_ch, From ab07dd227398c6a3decf283b20ecea4c01f6d4ef Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Wed, 27 Aug 2025 11:09:46 +0200 Subject: [PATCH 230/317] second part for multiple samples processing --- conf/modules.config | 15 ++++++++++----- workflows/dmscore.nf | 26 +++++++++++++------------- 2 files changed, 23 insertions(+), 18 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 73b499e..755c9cc 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -65,11 +65,16 @@ process { } withName: 'GATK_SATURATIONMUTAGENESIS' { - publishDir = [ - path: "${params.outdir}/intermediate_files/gatk", - mode: 'copy', - saveAs: { filename -> filename == 'versions.yml' ? null : filename } - ] + publishDir = [ + path: "${params.outdir}/intermediate_files/gatk", + mode: 'copy', + overwrite: false, + // put everything except versions.yml under a folder named by the sample id + saveAs: { filename -> + if (filename == 'versions.yml') return null + "${meta.id}/${filename}" + } + ] } withName: 'DMSANALYSIS_POSSIBLE_MUTATIONS' { diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index 68e5447..ffd0a36 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -222,19 +222,19 @@ workflow DMSCORE { ch_fasta_path_broadcast // path(fasta) ) -// FASTA path for GATK: broadcast to N -ch_fasta_for_gatk = ch_fasta.combine(PREMERGE.out.bam).map { it[1] } // path -- N -// Reading frame for GATK: broadcast to N (it's a val string) -ch_rf_for_gatk = reading_frame_ch.combine(PREMERGE.out.bam).map { it[0] } // val -- N -// min_counts for GATK: broadcast to N (also a val) -ch_min_for_gatk = min_counts_ch.combine(PREMERGE.out.bam).map { it[0] } // val -- N - -GATK_SATURATIONMUTAGENESIS( - PREMERGE.out.bam, // merged reads - tuple(val(meta), path(bam)) - ch_fasta_for_gatk, // path(fasta) - ch_rf_for_gatk, // val(reading_frame string) - ch_min_for_gatk // val(min_counts) -) + // FASTA path for GATK: broadcast to N + ch_fasta_for_gatk = ch_fasta.combine(PREMERGE.out.bam).map { it[1] } // path -- N + // Reading frame for GATK: broadcast to N (it's a val string) + ch_rf_for_gatk = reading_frame_ch.combine(PREMERGE.out.bam).map { it[0] } // val -- N + // min_counts for GATK: broadcast to N (also a val) + ch_min_for_gatk = min_counts_ch.combine(PREMERGE.out.bam).map { it[0] } // val -- N + + GATK_SATURATIONMUTAGENESIS( + PREMERGE.out.bam, // merged reads - tuple(val(meta), path(bam)) + ch_fasta_for_gatk, // path(fasta) + ch_rf_for_gatk, // val(reading_frame string) + ch_min_for_gatk // val(min_counts) + ) DMSANALYSIS_AASEQ ( ch_fasta, From 6dd6255e0b39e52af4422dfa06d6455220f44043 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Wed, 27 Aug 2025 12:28:18 +0200 Subject: [PATCH 231/317] third part for multiple samples processing --- conf/modules.config | 54 +++---------- modules/local/gatk/gatktodimsum.nf | 6 +- workflows/dmscore.nf | 122 ++++++++++++++++++++--------- 3 files changed, 95 insertions(+), 87 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 755c9cc..0aa87f2 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -101,53 +101,17 @@ process { ] } - withName: 'VISUALIZATION_COUNTS_PER_COV' { + withName: /.*VISUALIZATION_.*/ { publishDir = [ - path: "${params.outdir}/final_plots", - mode: 'copy', - saveAs: { filename -> filename == 'versions.yml' ? null : filename } + path: { "${params.outdir}/final_plots" }, // e.g. results/final_plots + mode: 'copy', + overwrite: false, + saveAs: { fn -> + if (fn == 'versions.yml') return null + "${meta.id}/${fn}" // put every output under the sample's subfolder + } ] - } - - withName: 'VISUALIZATION_COUNTS_HEATMAP' { - publishDir = [ - path: "${params.outdir}/final_plots", - mode: 'copy', - saveAs: { filename -> filename == 'versions.yml' ? null : filename } - ] - } - - withName: 'VISUALIZATION_GLOBAL_POS_BIASES_COUNTS' { - publishDir = [ - path: "${params.outdir}/final_plots", - mode: 'copy', - saveAs: { filename -> filename == 'versions.yml' ? null : filename } - ] - } - - withName: 'VISUALIZATION_GLOBAL_POS_BIASES_COV' { - publishDir = [ - path: "${params.outdir}/final_plots", - mode: 'copy', - saveAs: { filename -> filename == 'versions.yml' ? null : filename } - ] - } - - withName: 'VISUALIZATION_LOGDIFF' { - publishDir = [ - path: "${params.outdir}/final_plots", - mode: 'copy', - saveAs: { filename -> filename == 'versions.yml' ? null : filename } - ] - } - - withName: 'VISUALIZATION_SEQDEPTH' { - publishDir = [ - path: "${params.outdir}/final_plots", - mode: 'copy', - saveAs: { filename -> filename == 'versions.yml' ? null : filename } - ] - } + } withName: 'GATK_GATKTODIMSUM' { publishDir = [ diff --git a/modules/local/gatk/gatktodimsum.nf b/modules/local/gatk/gatktodimsum.nf index 9c51a88..f23335f 100644 --- a/modules/local/gatk/gatktodimsum.nf +++ b/modules/local/gatk/gatktodimsum.nf @@ -14,7 +14,7 @@ process GATK_GATKTODIMSUM { path script // R script output: - tuple val(meta), path("dimsum_input.tsv"), emit: dimsum_input + tuple val(meta), path("${meta.id}_dimsum_input.tsv"), emit: dimsum_input path "versions.yml", emit: versions when: @@ -24,7 +24,7 @@ process GATK_GATKTODIMSUM { """ start_stop_codon="$pos_range" - Rscript -e "source('$script'); generate_dimsum_input('$wt_seq', '$variantCounts_filtered_by_library', '\$start_stop_codon', 'dimsum_input.tsv')" + Rscript -e "source('$script'); generate_dimsum_input('$wt_seq', '$variantCounts_filtered_by_library', '\$start_stop_codon', '${meta.id}_dimsum_input.tsv')" R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') cat <<-END_VERSIONS > versions.yml @@ -35,7 +35,7 @@ process GATK_GATKTODIMSUM { stub: """ - touch dimsum_input.tsv + touch ${meta.id}_dimsum_input.tsv echo "GATK_GATKTODIMSUM:" > versions.yml echo " stub-version: 0.0.0" >> versions.yml """ diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index ffd0a36..c723a13 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -183,9 +183,6 @@ workflow DMSCORE { // Broadcast index to all samples ch_bwa_index = BWA_INDEX.out.index - - - // Broadcast the index to all samples ch_bwa_index_broadcast = ch_samplesheet .combine(ch_bwa_index) @@ -207,7 +204,6 @@ workflow DMSCORE { ch_sort_bam ) - BAMFILTER_DMS ( BWA_MEM.out.bam ) @@ -252,68 +248,116 @@ workflow DMSCORE { ) ch_versions = ch_versions.mix(DMSANALYSIS_POSSIBLE_MUTATIONS.out.versions) + // Anchor (N items; one per sample) + def ch_vc = GATK_SATURATIONMUTAGENESIS.out.variantCounts // tuple(val(meta), path) + + // Fan-out helpers (broadcast singleton → N) + def fanout = { ch_singleton -> ch_singleton.combine(ch_vc).map { it[0] } } + + // Build per-sample inputs + ch_possible_mut_for_proc = fanout( DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations.map { it[1] } ) + ch_aa_seq_for_proc = fanout( DMSANALYSIS_AASEQ.out.aa_seq.map { it[1] } ) + ch_min_counts_for_proc = fanout( min_counts_ch ) + ch_proc_raw_script = fanout( process_raw_gatk_script_ch ) + ch_filter_lib_script = fanout( filter_by_library_script_ch ) + ch_complete_script = fanout( complete_gatk_script_ch ) + ch_prepare_heatmap_script= fanout( prepare_counts_heatmap_script_ch ) + + // Call with all inputs aligned (each has N items now) DMSANALYSIS_PROCESS_GATK( - GATK_SATURATIONMUTAGENESIS.out.variantCounts, - DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations.map{ it[1] }, - DMSANALYSIS_AASEQ.out.aa_seq.map{ it[1] }, - min_counts_ch, - process_raw_gatk_script_ch, - filter_by_library_script_ch, - complete_gatk_script_ch, - prepare_counts_heatmap_script_ch - ) + ch_vc, // tuple(val(meta), path(variantCounts)) -- N + ch_possible_mut_for_proc, // path(possible_mutations) -- N + ch_aa_seq_for_proc, // path(aa_seq) -- N + ch_min_counts_for_proc, // val(min_counts) -- N + ch_proc_raw_script, // path(R script) -- N + ch_filter_lib_script, // path(R script) -- N + ch_complete_script, // path(R script) -- N + ch_prepare_heatmap_script // path(R script) -- N + ) annotated_variantCounts_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, a) } variantCounts_filtered_by_library_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, b) } library_completed_variantCounts_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, c) } variantCounts_for_heatmaps_ch = DMSANALYSIS_PROCESS_GATK.out.processed_variantCounts.map { meta, a, b, c, d -> tuple(meta, d) } + // Broadcast `singleton` so it emits once per item in `anchorN` + def fanoutTo = { anchorN, singleton -> singleton.combine(anchorN).map { it[0] } } + + // --- For VISUALIZATION_COUNTS_PER_COV (anchor: variantCounts_for_heatmaps_ch) + min_counts_for_cov_ch = fanoutTo(variantCounts_for_heatmaps_ch, min_counts_ch) + counts_per_cov_heatmap_scriptN = fanoutTo(variantCounts_for_heatmaps_ch, counts_per_cov_heatmap_script_ch) + + // --- For VISUALIZATION_COUNTS_HEATMAP (anchor: variantCounts_for_heatmaps_ch) + min_counts_for_heatmap_ch = fanoutTo(variantCounts_for_heatmaps_ch, min_counts_ch) + counts_heatmap_scriptN = fanoutTo(variantCounts_for_heatmaps_ch, counts_heatmap_script_ch) + + // --- For VISUALIZATION_GLOBAL_POS_BIASES_* (anchor: variantCounts_filtered_by_library_ch) + aa_seq_for_bias_ch = fanoutTo(variantCounts_filtered_by_library_ch, DMSANALYSIS_AASEQ.out.aa_seq.map { it[1] }) + sliding_window_size_N = fanoutTo(variantCounts_filtered_by_library_ch, sliding_window_size_ch) + aimed_cov_N = fanoutTo(variantCounts_filtered_by_library_ch, aimed_cov_ch) + global_bias_counts_cov_scriptN = fanoutTo(variantCounts_filtered_by_library_ch, global_bias_counts_cov_script_ch) + global_bias_cov_scriptN = fanoutTo(variantCounts_filtered_by_library_ch, global_bias_cov_script_ch) + + // --- For VISUALIZATION_LOGDIFF (anchor: library_completed_variantCounts_ch) +logdiff_scriptN = fanoutTo(library_completed_variantCounts_ch, logdiff_script_ch) + + // --- For VISUALIZATION_SEQDEPTH (anchor: variantCounts_filtered_by_library_ch) + possible_mutations_N = fanoutTo(variantCounts_filtered_by_library_ch, DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations.map { it[1] }) + min_counts_for_seqdepth_ch = fanoutTo(variantCounts_filtered_by_library_ch, min_counts_ch) + seqdepth_simulation_scriptN = fanoutTo(variantCounts_filtered_by_library_ch, seqdepth_simulation_script_ch) + VISUALIZATION_COUNTS_PER_COV( - variantCounts_for_heatmaps_ch, - min_counts_ch, - counts_per_cov_heatmap_script_ch + variantCounts_for_heatmaps_ch, + min_counts_for_cov_ch, + counts_per_cov_heatmap_scriptN ) VISUALIZATION_COUNTS_HEATMAP( - variantCounts_for_heatmaps_ch, - min_counts_ch, - counts_heatmap_script_ch + variantCounts_for_heatmaps_ch, + min_counts_for_heatmap_ch, + counts_heatmap_scriptN ) VISUALIZATION_GLOBAL_POS_BIASES_COUNTS( - variantCounts_filtered_by_library_ch, - DMSANALYSIS_AASEQ.out.aa_seq.map{ it[1] }, - sliding_window_size_ch, - global_bias_counts_cov_script_ch + variantCounts_filtered_by_library_ch, + aa_seq_for_bias_ch, + sliding_window_size_N, + global_bias_counts_cov_scriptN ) VISUALIZATION_GLOBAL_POS_BIASES_COV( - variantCounts_filtered_by_library_ch, - DMSANALYSIS_AASEQ.out.aa_seq.map{ it[1] }, - sliding_window_size_ch, - aimed_cov_ch, - global_bias_cov_script_ch + variantCounts_filtered_by_library_ch, + aa_seq_for_bias_ch, + sliding_window_size_N, + aimed_cov_N, + global_bias_cov_scriptN ) VISUALIZATION_LOGDIFF( - library_completed_variantCounts_ch, - logdiff_script_ch + library_completed_variantCounts_ch, + logdiff_scriptN ) if (params.run_seqdepth) { - VISUALIZATION_SEQDEPTH( + VISUALIZATION_SEQDEPTH( variantCounts_filtered_by_library_ch, - DMSANALYSIS_POSSIBLE_MUTATIONS.out.possible_mutations.map{ it[1] }, - min_counts_ch, - seqdepth_simulation_script_ch - ) + possible_mutations_N, + min_counts_for_seqdepth_ch, + seqdepth_simulation_scriptN + ) } + // Broadcast singletons to N (one per sample), anchored on variantCounts_filtered_by_library_ch + ch_fasta_for_dimsum = ch_fasta.combine(variantCounts_filtered_by_library_ch).map { it[1] } // path(fasta) -- N + ch_rf_for_dimsum = reading_frame_ch.combine(variantCounts_filtered_by_library_ch).map { it[0] } // val(range) -- N + ch_script_for_dimsum = gatk_to_dimsum_script_ch.combine(variantCounts_filtered_by_library_ch).map { it[0] } // path(script) -- N + + // Call with aligned inputs GATK_GATKTODIMSUM( - variantCounts_filtered_by_library_ch, - ch_fasta.map{ it[1] }, - reading_frame_ch, - gatk_to_dimsum_script_ch + variantCounts_filtered_by_library_ch, // tuple(val(meta), path) + ch_fasta_for_dimsum, // path(fasta) + ch_rf_for_dimsum, // val(reading_frame) + ch_script_for_dimsum // path(R script) ) emit: From ba59f7c7e4e379bc1c787ff400fb8ee799fee5db Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Wed, 27 Aug 2025 15:13:44 +0200 Subject: [PATCH 232/317] Create pipeline_steps.md --- docs/pipeline_steps.md | 160 +++++++++++++++++++++++++++++++++++++++++ 1 file changed, 160 insertions(+) create mode 100644 docs/pipeline_steps.md diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md new file mode 100644 index 0000000..dc04a6f --- /dev/null +++ b/docs/pipeline_steps.md @@ -0,0 +1,160 @@ +# nf-core/dms: Detailed Pipeline Steps + +This page provides an in-depth description of the data processing +modules implemented in **nf-core/dms**.\ +It is intended for advanced users and developers who want to understand +the rationale behind design choices, explore implementation details, and +consider potential future extensions. + +------------------------------------------------------------------------ + +## Overview + +The pipeline processes deep mutational scanning (DMS) sequencing data in +several stages:\ +1. Alignment of reads to the reference ORF\ +2. Filtering of wildtype and erroneous reads\ +3. Read merging for error reduction\ +4. Mutation counting\ +5. DMS library quality control\ +6. Data summarisation across samples\ +7. Single nucleotide variant error correction *(in development)*\ +8. Fitness estimation *(in development)* + +Each step is explained below. Links are provided to the primary tools +and libraries used, where applicable. + +------------------------------------------------------------------------ + +## 1. Alignment + +All paired-end raw reads are aligned to the reference ORF using +[**bwa-mem**](http://bio-bwa.sourceforge.net/).\ +This is a highly efficient mapping algorithm for reads ≥ 50 bp, with its +multi-threading support automatically handled by nf-core. + +In future versions of this pipeline, we consider the use of +[**bwa-mem2**](https://github.com/bwa-mem2/bwa-mem2), which provides +similar alignment rates with a moderate speed increase (preprint).\ +With the increasing diversity of sequencing platforms for DMS, new +throughput, read length, and error profiles may require further +alignment options to be implemented. + +------------------------------------------------------------------------ + +## 2. Filtering + +For long ORF site-saturation mutagenesis libraries, most aligned shotgun +sequencing reads contain exact matches against the reference.\ +It is not possible to infer which of these stem from mutant versus +wildtype DNA molecules prior to fragmentation, hence they are filtered +out. + +Similarly, erroneous reads with unexpected indels are also removed.\ +To this end, we use a custom Python script that relies on the +[**pysam**](https://pysam.readthedocs.io/en/latest/) library. + +------------------------------------------------------------------------ + +## 3. Read Merging + +Even the highest-accuracy next-generation sequencing platforms do not +have perfect base accuracy.\ +To minimise the effect of base errors (which would otherwise be counted +as "false mutations"), nf-core/dms uses the overlap of each aligned read +pair. + +With base errors on the forward and reverse read being independent, the +pipeline applies the [**vsearch +mergereads**](https://github.com/torognes/vsearch) function to convert +each read pair into a single consensus molecule with adjusted base error +scores. + +> **Tip**: Optimal merging performance is usually obtained if the +> average DNA fragment size matches the read size. For example, +> libraries sequenced with 150 bp paired-end reads should ideally also +> be sheared/tagmented to a mean size of 150 bp. + +Future versions may offer additional options depending on sequencing +type and error profiles. + +------------------------------------------------------------------------ + +## 4. Mutation Counting + +Aligned, non-wildtype consensus reads are screened for exact, base-level +mismatches.\ +nf-core/dms uses a custom Python implementation to count the occurrences +of all high-quality single, double, triple, and higher-order nucleotide +changes between each read and the reference ORF. + +Unlike the popular [**GATK +AnalyzeSaturationMutagenesis**](https://gatk.broadinstitute.org/hc/en-us/articles/360037057952-AnalyzeSaturationMutagenesis), +nf-core/dms allows users to specify a minimum base quality cutoff for +mutations to be included in the final count table (default: Q30). + +------------------------------------------------------------------------ + +## 5. DMS Library Quality Control + +By integrating the reference ORF coordinates and the chosen DMS library +type (default: NNK/NNs degenerate codon-based nicking), nf-core/dms +calculates a number of mutation count summary statistics. + +Custom visualisations allow for inspection of: 1. Mutation efficiency +along the ORF\ +2. Position-specific recovery of amino acid diversity\ +3. Overall sequencing coverage evenness and saturation + +------------------------------------------------------------------------ + +## 6. Data Summarisation for Fitness Estimation + +Steps 1--5 are iteratively run across all samples defined in the `.csv` +spreadsheet.\ +Once read alignment, merging, mutation counting, and library QC have +been completed for the full list of samples, users can opt to proceed +with fitness estimation using a number of tools (default: +[**DiMSum**](https://github.com/lehner-lab/DiMSum)). + +The pipeline generates the tool-specific input files by merging mutation +counts across samples. + +------------------------------------------------------------------------ + +## 7. Single Nucleotide Variant Error Correction *(in development)* + +This module will implement strategies to distinguish true low-frequency +variants from sequencing artefacts.\ +Methods under consideration include:\ +- Empirical error rate modelling based on wildtype controls\ +- Position-specific background correction\ +- Integration of UMIs (unique molecular identifiers), where available + +------------------------------------------------------------------------ + +## 8. Fitness Estimation *(in development)* + +The final step of the pipeline will perform fitness estimation based on +mutation counts. Planned features include:\ +- Integration of multiple fitness inference tools beyond DiMSum\ +- Options for replicates and condition-specific comparisons\ +- Standardised output formats for downstream analysis and visualisation + +------------------------------------------------------------------------ + +## Notes for Developers + +- Custom scripts used in filtering and mutation counting are available + in the `bin/` directory of the repository.\ +- Modules are implemented in Nextflow DSL2 and follow the nf-core + community guidelines.\ +- Contributions, optimisations, and additional analysis modules are + welcome --- please open a pull request or GitHub issue to discuss + ideas. + +------------------------------------------------------------------------ + +*This document is meant as a living reference. As the pipeline evolves, +the descriptions of steps 7 and 8 will be expanded with concrete +implementation details.* From bc608968a38d340b0ea8b4aedf51fea24c472014 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Wed, 27 Aug 2025 15:14:42 +0200 Subject: [PATCH 233/317] Update pipeline_steps.md --- docs/pipeline_steps.md | 6 +----- 1 file changed, 1 insertion(+), 5 deletions(-) diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md index dc04a6f..5f46b43 100644 --- a/docs/pipeline_steps.md +++ b/docs/pipeline_steps.md @@ -1,10 +1,6 @@ # nf-core/dms: Detailed Pipeline Steps -This page provides an in-depth description of the data processing -modules implemented in **nf-core/dms**.\ -It is intended for advanced users and developers who want to understand -the rationale behind design choices, explore implementation details, and -consider potential future extensions. +This page provides an in-depth description of the data processing modules implemented in **nf-core/dms**. It is intended for advanced users and developers who want to understand the rationale behind design choices, explore implementation details, and consider potential future extensions. ------------------------------------------------------------------------ From 5b559195332a362a1e9e30b34ddc81a57749d409 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Wed, 27 Aug 2025 15:16:34 +0200 Subject: [PATCH 234/317] lowcpu profile --- nextflow.config | 17 +++++++++++++++++ 1 file changed, 17 insertions(+) diff --git a/nextflow.config b/nextflow.config index 585ddad..7b821d2 100644 --- a/nextflow.config +++ b/nextflow.config @@ -194,6 +194,23 @@ profiles { } } } + lowcpu { + process { + // default for all processes when this profile is active + cpus = 1 + + // override resource labels defined (e.g., in base.config) + withLabel: process_single { cpus = 1 } + withLabel: process_low { cpus = 1 } + withLabel: process_medium { cpus = 1 } + withLabel: process_high { cpus = 1 } + withLabel: process_long { cpus = 1 } + withLabel: process_high_memory { cpus = 1 } + + // anything not labeled gets also 1 CPU + withName: /.*/ { cpus = 1 } + } + } test { includeConfig 'conf/test.config' } test_full { includeConfig 'conf/test_full.config' } } From 8a950fffde6bf7120c3fe1807b6fff2dfdd81229 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Wed, 27 Aug 2025 15:36:22 +0200 Subject: [PATCH 235/317] launch from outside the folder --- workflows/dmscore.nf | 41 +++++++++++++++++++++++------------------ 1 file changed, 23 insertions(+), 18 deletions(-) diff --git a/workflows/dmscore.nf b/workflows/dmscore.nf index c723a13..0a88dbe 100644 --- a/workflows/dmscore.nf +++ b/workflows/dmscore.nf @@ -83,24 +83,29 @@ Channel // Define R scripts as channels -Channel.fromPath("modules/local/dmsanalysis/bin/SeqDepth_simulation.R", checkIfExists: true).set { seqdepth_simulation_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/aa_seq.R", checkIfExists: true).set { aa_seq_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/complete_prefiltered_gatk.R", checkIfExists: true).set { complete_gatk_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/counts_heatmap.R", checkIfExists: true).set { counts_heatmap_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R", checkIfExists: true).set { counts_per_cov_heatmap_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/detect_codons.R", checkIfExists: true).set { detect_codons_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/filter_gatk_by_codon_library.R", checkIfExists: true).set { filter_by_library_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/fitness_heatmap.R", checkIfExists: true).set { fitness_heatmap_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/gatk_to_dimsum.R", checkIfExists: true).set { gatk_to_dimsum_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/global_position_biases_counts_and_counts_per_cov.R", checkIfExists: true).set { global_bias_counts_cov_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/global_position_biases_cov.R", checkIfExists: true).set { global_bias_cov_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/install_packages.R", checkIfExists: true).set { install_packages_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/logdiff.R", checkIfExists: true).set { logdiff_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/low_count_variants.R", checkIfExists: true).set { low_count_variants_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/possible_mutations.R", checkIfExists: true).set { possible_mutations_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/prepare_gatk_data_for_count_heatmaps.R", checkIfExists: true).set { prepare_counts_heatmap_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R", checkIfExists: true).set { prepare_fitness_heatmap_script_ch } -Channel.fromPath("modules/local/dmsanalysis/bin/process_raw_gatk.R", checkIfExists: true).set { process_raw_gatk_script_ch } +// Helper to anchor to the repo root +def R = { rel -> Channel.fromPath("${workflow.projectDir}/${rel}", checkIfExists: true) } + +// Define R scripts as channels (repo-anchored, works from any launch dir) +R("modules/local/dmsanalysis/bin/SeqDepth_simulation.R").set { seqdepth_simulation_script_ch } +R("modules/local/dmsanalysis/bin/aa_seq.R").set { aa_seq_script_ch } +R("modules/local/dmsanalysis/bin/complete_prefiltered_gatk.R").set { complete_gatk_script_ch } +R("modules/local/dmsanalysis/bin/counts_heatmap.R").set { counts_heatmap_script_ch } +R("modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R").set { counts_per_cov_heatmap_script_ch } +R("modules/local/dmsanalysis/bin/detect_codons.R").set { detect_codons_script_ch } +R("modules/local/dmsanalysis/bin/filter_gatk_by_codon_library.R").set { filter_by_library_script_ch } +R("modules/local/dmsanalysis/bin/fitness_heatmap.R").set { fitness_heatmap_script_ch } +R("modules/local/dmsanalysis/bin/gatk_to_dimsum.R").set { gatk_to_dimsum_script_ch } +R("modules/local/dmsanalysis/bin/global_position_biases_counts_and_counts_per_cov.R").set { global_bias_counts_cov_script_ch } +R("modules/local/dmsanalysis/bin/global_position_biases_cov.R").set { global_bias_cov_script_ch } +R("modules/local/dmsanalysis/bin/install_packages.R").set { install_packages_script_ch } +R("modules/local/dmsanalysis/bin/logdiff.R").set { logdiff_script_ch } +R("modules/local/dmsanalysis/bin/low_count_variants.R").set { low_count_variants_script_ch } +R("modules/local/dmsanalysis/bin/possible_mutations.R").set { possible_mutations_script_ch } +R("modules/local/dmsanalysis/bin/prepare_gatk_data_for_count_heatmaps.R").set { prepare_counts_heatmap_script_ch } +R("modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R").set { prepare_fitness_heatmap_script_ch } +R("modules/local/dmsanalysis/bin/process_raw_gatk.R").set { process_raw_gatk_script_ch } + workflow DMSCORE { From cb2624a1266aa7523429d1049c1a17c5393146a4 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Wed, 27 Aug 2025 15:37:37 +0200 Subject: [PATCH 236/317] Update pipeline_steps.md --- docs/pipeline_steps.md | 133 +++++++++++++---------------------------- 1 file changed, 40 insertions(+), 93 deletions(-) diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md index 5f46b43..d3ee1c8 100644 --- a/docs/pipeline_steps.md +++ b/docs/pipeline_steps.md @@ -1,156 +1,103 @@ # nf-core/dms: Detailed Pipeline Steps -This page provides an in-depth description of the data processing modules implemented in **nf-core/dms**. It is intended for advanced users and developers who want to understand the rationale behind design choices, explore implementation details, and consider potential future extensions. +This page provides in-depth descriptions of the data processing modules implemented in **nf-core/dms**. It is mainly intended for advanced users and developers who want to understand the rationale behind design choices, explore implementation details, and consider potential future extensions. ------------------------------------------------------------------------ ## Overview -The pipeline processes deep mutational scanning (DMS) sequencing data in -several stages:\ -1. Alignment of reads to the reference ORF\ -2. Filtering of wildtype and erroneous reads\ -3. Read merging for error reduction\ -4. Mutation counting\ -5. DMS library quality control\ -6. Data summarisation across samples\ -7. Single nucleotide variant error correction *(in development)*\ +The pipeline processes deep mutational scanning (DMS) sequencing data in several stages: +1. Alignment of reads to the reference open reading frame (ORF) +2. Filtering of wildtype and erroneous reads +3. Read merging for base error reduction +4. Mutation counting +5. DMS library quality control +6. Data summarisation across samples +7. Single nucleotide variant error correction *(in development)* 8. Fitness estimation *(in development)* -Each step is explained below. Links are provided to the primary tools -and libraries used, where applicable. +Each step is explained below. Links are provided to the primary tools and libraries used, where applicable. ------------------------------------------------------------------------ ## 1. Alignment -All paired-end raw reads are aligned to the reference ORF using -[**bwa-mem**](http://bio-bwa.sourceforge.net/).\ -This is a highly efficient mapping algorithm for reads ≥ 50 bp, with its -multi-threading support automatically handled by nf-core. +All paired-end raw reads are first aligned to the provided reference ORF using [**bwa-mem**](http://bio-bwa.sourceforge.net/). This is a highly efficient mapping algorithm for reads ≥100 bp, with its multi-threading support automatically handled by nf-core. -In future versions of this pipeline, we consider the use of -[**bwa-mem2**](https://github.com/bwa-mem2/bwa-mem2), which provides -similar alignment rates with a moderate speed increase (preprint).\ -With the increasing diversity of sequencing platforms for DMS, new -throughput, read length, and error profiles may require further -alignment options to be implemented. +In future versions of nf-core/dms, we consider the use of [**bwa-mem2**](https://github.com/bwa-mem2/bwa-mem2), which provides similar alignment rates with a moderate speed increase ([Vasimuddin et al., *IPDPS* 2019](https://ieeexplore.ieee.org/document/8820962). With the increasing diversity of sequencing platforms for DMS, new throughput, read length, and error profiles may require further alignment options to be implemented. ------------------------------------------------------------------------ ## 2. Filtering -For long ORF site-saturation mutagenesis libraries, most aligned shotgun -sequencing reads contain exact matches against the reference.\ -It is not possible to infer which of these stem from mutant versus -wildtype DNA molecules prior to fragmentation, hence they are filtered -out. +For long ORF site-saturation mutagenesis libraries, most aligned shotgun sequencing reads contain exact matches against the reference. It is not possible to infer which of these stem from mutant versus wildtype DNA molecules prior to fragmentation, hence they are filtered out. Similarly, erroneous reads with unexpected indels are also removed. -Similarly, erroneous reads with unexpected indels are also removed.\ -To this end, we use a custom Python script that relies on the -[**pysam**](https://pysam.readthedocs.io/en/latest/) library. +To this end, we use [**samtools view**](https://www.htslib.org/doc/samtools.html). ------------------------------------------------------------------------ ## 3. Read Merging -Even the highest-accuracy next-generation sequencing platforms do not -have perfect base accuracy.\ -To minimise the effect of base errors (which would otherwise be counted -as "false mutations"), nf-core/dms uses the overlap of each aligned read -pair. +Even the highest-accuracy next-generation sequencing platforms do not have perfect base accuracy. To minimise the effect of base errors (which would otherwise be counted as "false mutations"), nf-core/dms uses the overlap of each aligned read pair. With base errors on the forward and reverse read being independent, the pipeline applies the [**vsearch fastq_mergepairs**](https://github.com/torognes/vsearch) function to convert each read pair into a single consensus molecule with adjusted base error scores. -With base errors on the forward and reverse read being independent, the -pipeline applies the [**vsearch -mergereads**](https://github.com/torognes/vsearch) function to convert -each read pair into a single consensus molecule with adjusted base error -scores. +> **Tip**: Optimal merging performance is usually obtained if the average DNA fragment size matches the read size. For example, libraries sequenced with 150 bp paired-end reads should ideally also be sheared/tagmented to a mean size of 150 bp. -> **Tip**: Optimal merging performance is usually obtained if the -> average DNA fragment size matches the read size. For example, -> libraries sequenced with 150 bp paired-end reads should ideally also -> be sheared/tagmented to a mean size of 150 bp. - -Future versions may offer additional options depending on sequencing -type and error profiles. +Future versions may offer additional options depending on sequencing type and error profiles. ------------------------------------------------------------------------ ## 4. Mutation Counting -Aligned, non-wildtype consensus reads are screened for exact, base-level -mismatches.\ -nf-core/dms uses a custom Python implementation to count the occurrences -of all high-quality single, double, triple, and higher-order nucleotide -changes between each read and the reference ORF. +Aligned, non-wildtype consensus reads are screened for exact, base-level mismatches. nf-core/dms uses a custom Python implementation to count the occurrences of all high-quality single, double, triple, and higher-order nucleotide changes between each read and the reference ORF. -Unlike the popular [**GATK -AnalyzeSaturationMutagenesis**](https://gatk.broadinstitute.org/hc/en-us/articles/360037057952-AnalyzeSaturationMutagenesis), -nf-core/dms allows users to specify a minimum base quality cutoff for -mutations to be included in the final count table (default: Q30). +Unlike the popular [**GATK AnalyzeSaturationMutagenesis**](https://gatk.broadinstitute.org/hc/en-us/articles/360037594771-AnalyzeSaturationMutagenesis-BETA) function, nf-core/dms allows users to specify a minimum base quality cutoff for mutations to be included in the final count table (default: Q30). ------------------------------------------------------------------------ ## 5. DMS Library Quality Control -By integrating the reference ORF coordinates and the chosen DMS library -type (default: NNK/NNs degenerate codon-based nicking), nf-core/dms -calculates a number of mutation count summary statistics. +By integrating the reference ORF coordinates and the chosen DMS library type (default: NNK/NNs degenerate codon-based nicking), nf-core/dms calculates a number of mutation count summary statistics. -Custom visualisations allow for inspection of: 1. Mutation efficiency -along the ORF\ -2. Position-specific recovery of amino acid diversity\ -3. Overall sequencing coverage evenness and saturation +Custom visualisations allow for inspection of (1) mutation efficiency along the ORF, (2) position-specific recovery of amino acid diversity, and (3) overall sequencing coverage evenness and saturation. ------------------------------------------------------------------------ ## 6. Data Summarisation for Fitness Estimation -Steps 1--5 are iteratively run across all samples defined in the `.csv` -spreadsheet.\ -Once read alignment, merging, mutation counting, and library QC have -been completed for the full list of samples, users can opt to proceed -with fitness estimation using a number of tools (default: -[**DiMSum**](https://github.com/lehner-lab/DiMSum)). - -The pipeline generates the tool-specific input files by merging mutation -counts across samples. +Steps 1-5 are iteratively run across all samples defined in the `.csv` spreadsheet. Once read alignment, merging, mutation counting, and library QC have been completed for the full list of samples, users can opt to proceed with fitness estimation using a number of tools (default: [**DiMSum**](https://github.com/lehner-lab/DiMSum)). The pipeline generates the tool-specific input files by merging mutation counts across samples, using custom R scripts. ------------------------------------------------------------------------ ## 7. Single Nucleotide Variant Error Correction *(in development)* -This module will implement strategies to distinguish true low-frequency -variants from sequencing artefacts.\ -Methods under consideration include:\ -- Empirical error rate modelling based on wildtype controls\ -- Position-specific background correction\ -- Integration of UMIs (unique molecular identifiers), where available +This module will implement strategies to distinguish true single nucleotide variants from sequencing artefacts. Methods under consideration include: +- Empirical error rate modelling based on false double mutants +- Empirical error rate modelling based on wildtype sequencing ------------------------------------------------------------------------ ## 8. Fitness Estimation *(in development)* -The final step of the pipeline will perform fitness estimation based on -mutation counts. Planned features include:\ -- Integration of multiple fitness inference tools beyond DiMSum\ -- Options for replicates and condition-specific comparisons\ -- Standardised output formats for downstream analysis and visualisation +The final step of the pipeline will perform fitness estimation based on mutation counts. Planned features include: +- Integration of multiple fitness inference tools beyond DiMSum, including [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) +- Options for replicates and condition-specific comparisons +- Standardised output formats for downstream analysis and comparison ------------------------------------------------------------------------ ## Notes for Developers -- Custom scripts used in filtering and mutation counting are available - in the `bin/` directory of the repository.\ -- Modules are implemented in Nextflow DSL2 and follow the nf-core - community guidelines.\ -- Contributions, optimisations, and additional analysis modules are - welcome --- please open a pull request or GitHub issue to discuss - ideas. +- Custom scripts used in filtering and mutation counting are available in the `bin/` directory of the repository. +- Modules are implemented in Nextflow DSL2 and follow the nf-core community guidelines. +- Contributions, optimisations, and additional analysis modules are welcome - please open a pull request or GitHub issue to discuss ideas. + +*This document is meant as a living reference. As the pipeline evolves, the descriptions of steps 7 and 8 will be expanded with concrete implementation details.* ------------------------------------------------------------------------ -*This document is meant as a living reference. As the pipeline evolves, -the descriptions of steps 7 and 8 will be expanded with concrete -implementation details.* +## Contact + +For detailled scientific or technical questions, feedback and experimental discussions, feel free to contact us directly: + +- Benjamin Wehnert — wehnertbenjamin@gmail.com +- Maximilian Stammnitz — maximilian.stammnitz@crg.eu From 80794fbd160cb659980144137e48cf4744b41bb7 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Wed, 27 Aug 2025 15:40:14 +0200 Subject: [PATCH 237/317] Update pipeline_steps.md --- docs/pipeline_steps.md | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md index d3ee1c8..8cead5c 100644 --- a/docs/pipeline_steps.md +++ b/docs/pipeline_steps.md @@ -16,6 +16,8 @@ The pipeline processes deep mutational scanning (DMS) sequencing data in several 7. Single nucleotide variant error correction *(in development)* 8. Fitness estimation *(in development)* +![pipeline](/docs/pipeline.png) + Each step is explained below. Links are provided to the primary tools and libraries used, where applicable. ------------------------------------------------------------------------ @@ -24,7 +26,7 @@ Each step is explained below. Links are provided to the primary tools and librar All paired-end raw reads are first aligned to the provided reference ORF using [**bwa-mem**](http://bio-bwa.sourceforge.net/). This is a highly efficient mapping algorithm for reads ≥100 bp, with its multi-threading support automatically handled by nf-core. -In future versions of nf-core/dms, we consider the use of [**bwa-mem2**](https://github.com/bwa-mem2/bwa-mem2), which provides similar alignment rates with a moderate speed increase ([Vasimuddin et al., *IPDPS* 2019](https://ieeexplore.ieee.org/document/8820962). With the increasing diversity of sequencing platforms for DMS, new throughput, read length, and error profiles may require further alignment options to be implemented. +In future versions of nf-core/dms, we consider the use of [**bwa-mem2**](https://github.com/bwa-mem2/bwa-mem2), which provides similar alignment rates with a moderate speed increase ([Vasimuddin et al., *IPDPS* 2019](https://ieeexplore.ieee.org/document/8820962)). 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z{XXh}-YVdyl!vc6j{_iBeR5XCBME+<^N4_-+BtH3BJQINT`Bc0uBv8v>XX=0?~_$! zOr2fNHQ`5`$L!BU#f-f!DQ;rNHnycY&rkGb+dg!}vidwm0+EqB+5 zh)mA-W5BpxPxH^$=krR7&rjv^>dfq1w4PfS9-6Xlt_~{e>Z|8{jf)*w9Dl35y^nGmHm|lNlTkQ9$=hEI~QK@$0NO!_~F=WQf8MFH# zSzK zam7t^2G@d$N8gW{U|cFDUSGpl7(VNL1CvTW?UA6^lvgRZ;=ZCjV~k6s3o)ozT66bN z{h;hfk0psdl2u(b7Qxjw;d_{OZ0b>{kk(vk(f3(3(rX1tl&j$;Rduo^%_GPk$ z%rPm~g>zS7;W)miT9}@^*yQ}6E$5~esb4Q<#g5+!ns9D;(SAO6)tW!*VwxBJYf#Co zh)TGjVoIALXFvVI$J-f1gHdQz(JAj8y%@BXMS9`~gRp~3m*761SM literal 0 HcmV?d00001 From 535a0d62d13cd8139780e92fc26a2fec1851f16f Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Wed, 27 Aug 2025 15:45:01 +0200 Subject: [PATCH 239/317] Update README.md --- README.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/README.md b/README.md index 9a868ed..babb12b 100644 --- a/README.md +++ b/README.md @@ -35,6 +35,8 @@ While DNA synthesis and sequencing technologies have advanced substantially, lon - Scalable across HPC and Cloud systems - Monitors CPU, memory, and CO₂ usage +For details of the pipeline and potential future expansions, please consider reading our [detailed description](docs/pipeline_steps.md). + --- ## 3. Installation From 2d4dad5d242bf9dced34577970c216da30c9d500 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Wed, 27 Aug 2025 22:57:05 +0200 Subject: [PATCH 240/317] Update pipeline_steps.md --- docs/pipeline_steps.md | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md index 8cead5c..2625248 100644 --- a/docs/pipeline_steps.md +++ b/docs/pipeline_steps.md @@ -42,7 +42,9 @@ To this end, we use [**samtools view**](https://www.htslib.org/doc/samtools.html Even the highest-accuracy next-generation sequencing platforms do not have perfect base accuracy. To minimise the effect of base errors (which would otherwise be counted as "false mutations"), nf-core/dms uses the overlap of each aligned read pair. With base errors on the forward and reverse read being independent, the pipeline applies the [**vsearch fastq_mergepairs**](https://github.com/torognes/vsearch) function to convert each read pair into a single consensus molecule with adjusted base error scores. -> **Tip**: Optimal merging performance is usually obtained if the average DNA fragment size matches the read size. For example, libraries sequenced with 150 bp paired-end reads should ideally also be sheared/tagmented to a mean size of 150 bp. +:::tip +Optimal merging performance is usually obtained if the average DNA fragment size matches the read size. For example, libraries sequenced with 150 bp paired-end reads should ideally also be sheared/tagmented to a mean size of 150 bp. +::: Future versions may offer additional options depending on sequencing type and error profiles. From 1126e41983f3309ceaa611aeee8cc24613d18463 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Wed, 27 Aug 2025 22:59:02 +0200 Subject: [PATCH 241/317] Update pipeline_steps.md --- docs/pipeline_steps.md | 5 ++--- 1 file changed, 2 insertions(+), 3 deletions(-) diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md index 2625248..b3db338 100644 --- a/docs/pipeline_steps.md +++ b/docs/pipeline_steps.md @@ -42,9 +42,8 @@ To this end, we use [**samtools view**](https://www.htslib.org/doc/samtools.html Even the highest-accuracy next-generation sequencing platforms do not have perfect base accuracy. To minimise the effect of base errors (which would otherwise be counted as "false mutations"), nf-core/dms uses the overlap of each aligned read pair. With base errors on the forward and reverse read being independent, the pipeline applies the [**vsearch fastq_mergepairs**](https://github.com/torognes/vsearch) function to convert each read pair into a single consensus molecule with adjusted base error scores. -:::tip -Optimal merging performance is usually obtained if the average DNA fragment size matches the read size. For example, libraries sequenced with 150 bp paired-end reads should ideally also be sheared/tagmented to a mean size of 150 bp. -::: +> [!TIP] +> Optimal merging performance is usually obtained if the average DNA fragment size matches the read size. For example, libraries sequenced with 150 bp paired-end reads should ideally also be sheared/tagmented to a mean size of 150 bp. Future versions may offer additional options depending on sequencing type and error profiles. From b25ec266f49e9def09206cb8f4816e29065de7a5 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 28 Aug 2025 13:52:23 +0200 Subject: [PATCH 242/317] Update README.md --- README.md | 16 ++++++++-------- 1 file changed, 8 insertions(+), 8 deletions(-) diff --git a/README.md b/README.md index babb12b..77512a0 100644 --- a/README.md +++ b/README.md @@ -18,15 +18,15 @@ --- ## 1. Overview -**nf-core/dms** is a reproducible, scalable, and community-curated pipeline for analyzing deep mutational scanning (DMS) data using shotgun DNA sequencing. DMS enables researchers to measure the fitness effects of thousands of gene variants simultaneously, helping to classify disease causing mutants in human and animal populations, to learn fundamental rules of virus evolution, protein architecture, splicing or small-molecule interactions. +**nf-core/deepmutscan** is a reproducible, scalable, and community-curated pipeline for analyzing deep mutational scanning (DMS) data using shotgun DNA sequencing. DMS enables researchers to measure the fitness effects of thousands of gene variants simultaneously, helping to classify disease causing mutants in human and animal populations, to learn fundamental rules of virus evolution, protein architecture, splicing or small-molecule interactions. -While DNA synthesis and sequencing technologies have advanced substantially, long open reading frame (ORF) targets still present major challenges for DMS studies. Shotgun DNA sequencing can be used to greatly speed up the inference of long ORF mutant fitness landscapes, theoretically at no expense in accuracy. We have designed the **nf-core/dms** pipeline to unlock the power of shotgun sequencing based DMS studies on long ORFs, to simplify and standardise the complex bioinformatics steps involved in data processing of such experiments – from read alignment to QC reporting and fitness landscape inferences. +While DNA synthesis and sequencing technologies have advanced substantially, long open reading frame (ORF) targets still present major challenges for DMS studies. Shotgun DNA sequencing can be used to greatly speed up the inference of long ORF mutant fitness landscapes, theoretically at no expense in accuracy. We have designed the **nf-core/deepmutscan** pipeline to unlock the power of shotgun sequencing based DMS studies on long ORFs, to simplify and standardise the complex bioinformatics steps involved in data processing of such experiments – from read alignment to QC reporting and fitness landscape inferences. > 📄 Reference: Wehnert et al., _bioRxiv_ preprint (coming soon) --- -## 2. Features of nf-core/dms +## 2. Features of nf-core/deepmutscan - End-to-end analyses of DMS shotgun sequencing data - Modular, three-stage workflow: alignment → QC → error-aware fitness estimation - Integrates with popular statistical tools like [DiMSum](https://github.com/lehner-lab/DiMSum), [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) @@ -40,7 +40,7 @@ For details of the pipeline and potential future expansions, please consider rea --- ## 3. Installation -**nf-core/dms** uses [Nextflow](https://nf-co.re/docs/usage/getting_started/installation), which must be installed on your system: +**nf-core/deepmutscan** uses [Nextflow](https://nf-co.re/docs/usage/getting_started/installation), which must be installed on your system: ```bash java -version # Check that Java v11+ is installed @@ -52,7 +52,7 @@ mv nextflow ~/bin/ # Add to user's $PATH The pipeline itself requires no installation – Nextflow will fetch it directly from GitHub: ```bash -nextflow run nf-core/dms -profile docker +nextflow run nf-core/deepmutscan -profile docker ``` --- @@ -62,10 +62,10 @@ Prepare: - A **sample sheet** CSV to specify input/output labels, replicates, etc. (see [example](assets/samplesheet.csv)) - A **reference FASTA** file for the gene or region of interest -To execute **nf-core/dms**, run the basic command: +To execute **nf-core/deepmutscan**, run the basic command: ```bash -nextflow run nf-core/dms \ +nextflow run nf-core/deepmutscan \ -profile singularity,local \ --input ./input.csv \ --outdir ./results \ @@ -151,7 +151,7 @@ We welcome contributions from the community! Please open an [issue](../../issues/new) or [pull request](../../compare) via this GitHub page, to: - Suggest or help implementing new modules for custom workflows -- Report bugs and other challenges in running **nf-core/dms** +- Report bugs and other challenges in running **nf-core/deepmutscan** - Help improve this documentation You can also reach out to us via the **nf-core Slack**, by use of the `#dms` channel ([join here](https://join.slack.com/share/enQtOTMyMDc3MTA0Mzg0Mi04YmRiNDEwZTBlOTRiN2M2ZGU5ZGVmOWQ3YzA0YjA4NzhiNjFhNTVlNDA4ZTZjOTE2MjE5MmIzYWZjZTljMTE3)). From 37d4e923874b122613adb1fc11c78c41dd7e856d Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 28 Aug 2025 13:54:29 +0200 Subject: [PATCH 243/317] Update pipeline_steps.md --- docs/pipeline_steps.md | 14 +++++++------- 1 file changed, 7 insertions(+), 7 deletions(-) diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md index b3db338..568b54e 100644 --- a/docs/pipeline_steps.md +++ b/docs/pipeline_steps.md @@ -1,6 +1,6 @@ -# nf-core/dms: Detailed Pipeline Steps +# nf-core/deepmutscan: Detailed Pipeline Steps -This page provides in-depth descriptions of the data processing modules implemented in **nf-core/dms**. It is mainly intended for advanced users and developers who want to understand the rationale behind design choices, explore implementation details, and consider potential future extensions. +This page provides in-depth descriptions of the data processing modules implemented in **nf-core/deepmutscan**. It is mainly intended for advanced users and developers who want to understand the rationale behind design choices, explore implementation details, and consider potential future extensions. ------------------------------------------------------------------------ @@ -26,7 +26,7 @@ Each step is explained below. Links are provided to the primary tools and librar All paired-end raw reads are first aligned to the provided reference ORF using [**bwa-mem**](http://bio-bwa.sourceforge.net/). This is a highly efficient mapping algorithm for reads ≥100 bp, with its multi-threading support automatically handled by nf-core. -In future versions of nf-core/dms, we consider the use of [**bwa-mem2**](https://github.com/bwa-mem2/bwa-mem2), which provides similar alignment rates with a moderate speed increase ([Vasimuddin et al., *IPDPS* 2019](https://ieeexplore.ieee.org/document/8820962)). With the increasing diversity of sequencing platforms for DMS, new throughput, read length, and error profiles may require further alignment options to be implemented. +In future versions of nf-core/deepmutscan, we consider the use of [**bwa-mem2**](https://github.com/bwa-mem2/bwa-mem2), which provides similar alignment rates with a moderate speed increase ([Vasimuddin et al., *IPDPS* 2019](https://ieeexplore.ieee.org/document/8820962)). With the increasing diversity of sequencing platforms for DMS, new throughput, read length, and error profiles may require further alignment options to be implemented. ------------------------------------------------------------------------ @@ -40,7 +40,7 @@ To this end, we use [**samtools view**](https://www.htslib.org/doc/samtools.html ## 3. Read Merging -Even the highest-accuracy next-generation sequencing platforms do not have perfect base accuracy. To minimise the effect of base errors (which would otherwise be counted as "false mutations"), nf-core/dms uses the overlap of each aligned read pair. With base errors on the forward and reverse read being independent, the pipeline applies the [**vsearch fastq_mergepairs**](https://github.com/torognes/vsearch) function to convert each read pair into a single consensus molecule with adjusted base error scores. +Even the highest-accuracy next-generation sequencing platforms do not have perfect base accuracy. To minimise the effect of base errors (which would otherwise be counted as "false mutations"), nf-core/deepmutscan uses the overlap of each aligned read pair. With base errors on the forward and reverse read being independent, the pipeline applies the [**vsearch fastq_mergepairs**](https://github.com/torognes/vsearch) function to convert each read pair into a single consensus molecule with adjusted base error scores. > [!TIP] > Optimal merging performance is usually obtained if the average DNA fragment size matches the read size. For example, libraries sequenced with 150 bp paired-end reads should ideally also be sheared/tagmented to a mean size of 150 bp. @@ -51,15 +51,15 @@ Future versions may offer additional options depending on sequencing type and er ## 4. Mutation Counting -Aligned, non-wildtype consensus reads are screened for exact, base-level mismatches. nf-core/dms uses a custom Python implementation to count the occurrences of all high-quality single, double, triple, and higher-order nucleotide changes between each read and the reference ORF. +Aligned, non-wildtype consensus reads are screened for exact, base-level mismatches. nf-core/deepmutscan uses a custom Python implementation to count the occurrences of all high-quality single, double, triple, and higher-order nucleotide changes between each read and the reference ORF. -Unlike the popular [**GATK AnalyzeSaturationMutagenesis**](https://gatk.broadinstitute.org/hc/en-us/articles/360037594771-AnalyzeSaturationMutagenesis-BETA) function, nf-core/dms allows users to specify a minimum base quality cutoff for mutations to be included in the final count table (default: Q30). +Unlike the popular [**GATK AnalyzeSaturationMutagenesis**](https://gatk.broadinstitute.org/hc/en-us/articles/360037594771-AnalyzeSaturationMutagenesis-BETA) function, nf-core/deepmutscan allows users to specify a minimum base quality cutoff for mutations to be included in the final count table (default: Q30). ------------------------------------------------------------------------ ## 5. DMS Library Quality Control -By integrating the reference ORF coordinates and the chosen DMS library type (default: NNK/NNs degenerate codon-based nicking), nf-core/dms calculates a number of mutation count summary statistics. +By integrating the reference ORF coordinates and the chosen DMS library type (default: NNK/NNs degenerate codon-based nicking), nf-core/deepmutscan calculates a number of mutation count summary statistics. Custom visualisations allow for inspection of (1) mutation efficiency along the ORF, (2) position-specific recovery of amino acid diversity, and (3) overall sequencing coverage evenness and saturation. From d272cfe64d75fb3cd96afd0b43c79fbd9021fea7 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Thu, 28 Aug 2025 14:39:51 +0200 Subject: [PATCH 244/317] multi-sample processing finalised --- conf/modules.config | 5 +- modules/local/gatk/annotategatk.nf | 91 ------------------------------ 2 files changed, 4 insertions(+), 92 deletions(-) delete mode 100644 modules/local/gatk/annotategatk.nf diff --git a/conf/modules.config b/conf/modules.config index 0aa87f2..132ccb7 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -97,7 +97,10 @@ process { publishDir = [ path: "${params.outdir}/intermediate_files/processed_gatk_files", mode: 'copy', - saveAs: { filename -> filename == 'versions.yml' ? null : filename } + saveAs: { filename -> + if (filename == 'versions.yml') return null + "${meta.id}/${filename}" + } ] } diff --git a/modules/local/gatk/annotategatk.nf b/modules/local/gatk/annotategatk.nf deleted file mode 100644 index eac3c50..0000000 --- a/modules/local/gatk/annotategatk.nf +++ /dev/null @@ -1,91 +0,0 @@ -// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) -// https://github.com/nf-core/modules/tree/master/modules/nf-core/ -// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: -// https://nf-co.re/join -// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. -// All other parameters MUST be provided using the "task.ext" directive, see here: -// https://www.nextflow.io/docs/latest/process.html#ext -// where "task.ext" is a string. -// Any parameters that need to be evaluated in the context of a particular sample -// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. -// TODO nf-core: Software that can be piped together SHOULD be added to separate module files -// unless there is a run-time, storage advantage in implementing in this way -// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: -// bwa mem | samtools view -B -T ref.fasta -// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty -// list (`[]`) instead of a file can be used to work around this issue. - -process DMSANALYSIS_ANNOTATEGATK { - tag "$meta.id" - label 'process_single' - - // TODO nf-core: List required Conda package(s). - // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). - // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. - // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. - conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/YOUR-TOOL-HERE': - 'biocontainers/YOUR-TOOL-HERE' }" - - input: - // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" - // MUST be provided as an input via a Groovy Map called "meta". - // This information may not be required in some instances e.g. indexing reference genome files: - // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf - // TODO nf-core: Where applicable please provide/convert compressed files as input/output - // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. - tuple val(meta), path(bam) - - output: - // TODO nf-core: Named file extensions MUST be emitted for ALL output channels - tuple val(meta), path("*.bam"), emit: bam - // TODO nf-core: List additional required output channels/values here - path "versions.yml" , emit: versions - - when: - task.ext.when == null || task.ext.when - - script: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 - // If the software is unable to output a version number on the command-line then it can be manually specified - // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf - // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) - // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive - // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter - // using the Nextflow "task" variable e.g. "--threads $task.cpus" - // TODO nf-core: Please replace the example samtools command below with your module's command - // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) - """ - samtools \\ - sort \\ - $args \\ - -@ $task.cpus \\ - -o ${prefix}.bam \\ - -T $prefix \\ - $bam - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') - END_VERSIONS - """ - - stub: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: A stub section should mimic the execution of the original module as best as possible - // Have a look at the following examples: - // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 - // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 - """ - touch ${prefix}.bam - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - dmsanalysis: \$(samtools --version |& sed '1!d ; s/samtools //') - END_VERSIONS - """ -} From 6b84cdc2119bad371c138172476a6a3ea9d318d4 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 28 Aug 2025 16:55:31 +0200 Subject: [PATCH 245/317] Update CONTRIBUTING.md --- .github/CONTRIBUTING.md | 12 ++++++------ 1 file changed, 6 insertions(+), 6 deletions(-) diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index 3a5f6ae..6c62cd8 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -1,22 +1,22 @@ -# `nf-core/dmscore`: Contributing Guidelines +# `nf-core/deepmutscan`: Contributing Guidelines Hi there! -Many thanks for taking an interest in improving nf-core/dmscore. +Many thanks for taking an interest in improving nf-core/deepmutscan. -We try to manage the required tasks for nf-core/dmscore using GitHub issues, you probably came to this page when creating one. +We try to manage the required tasks for nf-core/deepmutscan using GitHub issues, you probably came to this page when creating one. Please use the pre-filled template to save time. However, don't be put off by this template - other more general issues and suggestions are welcome! Contributions to the code are even more welcome ;) > [!NOTE] -> If you need help using or modifying nf-core/dmscore then the best place to ask is on the nf-core Slack [#dmscore](https://nfcore.slack.com/channels/dmscore) channel ([join our Slack here](https://nf-co.re/join/slack)). +> If you need help using or modifying nf-core/deepmutscan then the best place to ask is on the nf-core Slack [#deepmutscan](https://nfcore.slack.com/channels/dmscore) channel ([join our Slack here](https://nf-co.re/join/slack)). ## Contribution workflow -If you'd like to write some code for nf-core/dmscore, the standard workflow is as follows: +If you'd like to write some code for nf-core/deepmutscan, the standard workflow is as follows: -1. Check that there isn't already an issue about your idea in the [nf-core/dmscore issues](https://github.com/nf-core/dmscore/issues) to avoid duplicating work. If there isn't one already, please create one so that others know you're working on this +1. Check that there isn't already an issue about your idea in the [nf-core/deepmutscan issues](https://github.com/nf-core/dmscore/issues) to avoid duplicating work. If there isn't one already, please create one so that others know you're working on this 2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [nf-core/dmscore repository](https://github.com/nf-core/dmscore) to your GitHub account 3. Make the necessary changes / additions within your forked repository following [Pipeline conventions](#pipeline-contribution-conventions) 4. Use `nf-core pipelines schema build` and add any new parameters to the pipeline JSON schema (requires [nf-core tools](https://github.com/nf-core/tools) >= 1.10). From 3e0696ee950411ce62aea03da208b1b4e80a1d5a Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 28 Aug 2025 16:55:59 +0200 Subject: [PATCH 246/317] Update LICENSE --- LICENSE | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/LICENSE b/LICENSE index 48133aa..8119d7b 100644 --- a/LICENSE +++ b/LICENSE @@ -1,6 +1,6 @@ MIT License -Copyright (c) The nf-core/dmscore team +Copyright (c) The nf-core/deepmutscan team Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation files (the "Software"), to deal From 83d6e64b23b82dc41209c0c8974821aebfd1e684 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 28 Aug 2025 16:56:39 +0200 Subject: [PATCH 247/317] Update .nf-core.yml --- .nf-core.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.nf-core.yml b/.nf-core.yml index 59e948b..65d775f 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -6,7 +6,7 @@ lint: {} template: org: nf-core - name: dmscore + name: deepmutscan description: "Until now, most Deep Mutational Scanning (DMS) experiments relied\ \ on variant-specific barcoded libraries for sequencing. This method enabled DMS\ \ on large proteins and led to many great publications. Recently, efforts have\ From d71c2ad41db0d1a2dc8290212e937b19ff645fc2 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Thu, 28 Aug 2025 17:54:51 +0200 Subject: [PATCH 248/317] new name --- .nf-core.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.nf-core.yml b/.nf-core.yml index 59e948b..65d775f 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -6,7 +6,7 @@ lint: {} template: org: nf-core - name: dmscore + name: deepmutscan description: "Until now, most Deep Mutational Scanning (DMS) experiments relied\ \ on variant-specific barcoded libraries for sequencing. This method enabled DMS\ \ on large proteins and led to many great publications. Recently, efforts have\ From 7c4b2f7bef8f1bffdb9707f6d7a3d2567802ab77 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Thu, 28 Aug 2025 17:55:14 +0200 Subject: [PATCH 249/317] Template update for nf-core/tools version 3.1.2 --- .github/CONTRIBUTING.md | 20 ++++++------ .github/ISSUE_TEMPLATE/bug_report.yml | 4 +-- .github/ISSUE_TEMPLATE/config.yml | 6 ++-- .github/ISSUE_TEMPLATE/feature_request.yml | 2 +- .github/PULL_REQUEST_TEMPLATE.md | 10 +++--- .github/workflows/awsfulltest.yml | 6 ++-- .github/workflows/awstest.yml | 6 ++-- .github/workflows/branch.yml | 4 +-- .github/workflows/ci.yml | 2 +- .github/workflows/fix-linting.yml | 4 +-- .nf-core.yml | 19 +++++------ CHANGELOG.md | 4 +-- CITATIONS.md | 2 +- LICENSE | 2 +- README.md | 28 ++++++++-------- assets/adaptivecard.json | 2 +- assets/email_template.html | 14 ++++---- assets/email_template.txt | 10 +++--- assets/methods_description_template.yml | 8 ++--- assets/multiqc_config.yml | 8 ++--- assets/nf-core-deepmutscan_logo_light.png | Bin 0 -> 106639 bytes assets/nf-core-dmscore_logo_light.png | Bin 79195 -> 0 bytes assets/schema_input.json | 4 +-- assets/sendmail_template.txt | 6 ++-- assets/slackreport.json | 2 +- conf/base.config | 2 +- conf/test.config | 2 +- conf/test_full.config | 2 +- docs/README.md | 4 +-- docs/images/nf-core-deepmutscan_logo_dark.png | Bin 0 -> 28299 bytes .../images/nf-core-deepmutscan_logo_light.png | Bin 0 -> 23882 bytes docs/images/nf-core-dmscore_logo_dark.png | Bin 28505 -> 0 bytes docs/images/nf-core-dmscore_logo_light.png | Bin 24411 -> 0 bytes docs/output.md | 2 +- docs/usage.md | 12 +++---- main.nf | 26 +++++++-------- modules.json | 4 +-- nextflow.config | 16 +++++----- nextflow_schema.json | 6 ++-- ro-crate-metadata.json | 30 +++++++++--------- .../main.nf | 2 +- workflows/{dmscore.nf => deepmutscan.nf} | 6 ++-- 42 files changed, 142 insertions(+), 145 deletions(-) create mode 100644 assets/nf-core-deepmutscan_logo_light.png delete mode 100644 assets/nf-core-dmscore_logo_light.png create mode 100644 docs/images/nf-core-deepmutscan_logo_dark.png create mode 100644 docs/images/nf-core-deepmutscan_logo_light.png delete mode 100644 docs/images/nf-core-dmscore_logo_dark.png delete mode 100644 docs/images/nf-core-dmscore_logo_light.png rename subworkflows/local/{utils_nfcore_dmscore_pipeline => utils_nfcore_deepmutscan_pipeline}/main.nf (99%) rename workflows/{dmscore.nf => deepmutscan.nf} (96%) diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index 3a5f6ae..12dd4a0 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -1,23 +1,23 @@ -# `nf-core/dmscore`: Contributing Guidelines +# `nf-core/deepmutscan`: Contributing Guidelines Hi there! -Many thanks for taking an interest in improving nf-core/dmscore. +Many thanks for taking an interest in improving nf-core/deepmutscan. -We try to manage the required tasks for nf-core/dmscore using GitHub issues, you probably came to this page when creating one. +We try to manage the required tasks for nf-core/deepmutscan using GitHub issues, you probably came to this page when creating one. Please use the pre-filled template to save time. However, don't be put off by this template - other more general issues and suggestions are welcome! Contributions to the code are even more welcome ;) > [!NOTE] -> If you need help using or modifying nf-core/dmscore then the best place to ask is on the nf-core Slack [#dmscore](https://nfcore.slack.com/channels/dmscore) channel ([join our Slack here](https://nf-co.re/join/slack)). +> If you need help using or modifying nf-core/deepmutscan then the best place to ask is on the nf-core Slack [#deepmutscan](https://nfcore.slack.com/channels/deepmutscan) channel ([join our Slack here](https://nf-co.re/join/slack)). ## Contribution workflow -If you'd like to write some code for nf-core/dmscore, the standard workflow is as follows: +If you'd like to write some code for nf-core/deepmutscan, the standard workflow is as follows: -1. Check that there isn't already an issue about your idea in the [nf-core/dmscore issues](https://github.com/nf-core/dmscore/issues) to avoid duplicating work. If there isn't one already, please create one so that others know you're working on this -2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [nf-core/dmscore repository](https://github.com/nf-core/dmscore) to your GitHub account +1. Check that there isn't already an issue about your idea in the [nf-core/deepmutscan issues](https://github.com/nf-core/deepmutscan/issues) to avoid duplicating work. If there isn't one already, please create one so that others know you're working on this +2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [nf-core/deepmutscan repository](https://github.com/nf-core/deepmutscan) to your GitHub account 3. Make the necessary changes / additions within your forked repository following [Pipeline conventions](#pipeline-contribution-conventions) 4. Use `nf-core pipelines schema build` and add any new parameters to the pipeline JSON schema (requires [nf-core tools](https://github.com/nf-core/tools) >= 1.10). 5. Submit a Pull Request against the `dev` branch and wait for the code to be reviewed and merged @@ -61,11 +61,11 @@ These tests are run both with the latest available version of `Nextflow` and als ## Getting help -For further information/help, please consult the [nf-core/dmscore documentation](https://nf-co.re/dmscore/usage) and don't hesitate to get in touch on the nf-core Slack [#dmscore](https://nfcore.slack.com/channels/dmscore) channel ([join our Slack here](https://nf-co.re/join/slack)). +For further information/help, please consult the [nf-core/deepmutscan documentation](https://nf-co.re/deepmutscan/usage) and don't hesitate to get in touch on the nf-core Slack [#deepmutscan](https://nfcore.slack.com/channels/deepmutscan) channel ([join our Slack here](https://nf-co.re/join/slack)). ## Pipeline contribution conventions -To make the `nf-core/dmscore` code and processing logic more understandable for new contributors and to ensure quality, we semi-standardise the way the code and other contributions are written. +To make the `nf-core/deepmutscan` code and processing logic more understandable for new contributors and to ensure quality, we semi-standardise the way the code and other contributions are written. ### Adding a new step @@ -115,7 +115,7 @@ This repo includes a devcontainer configuration which will create a GitHub Codes To get started: -- Open the repo in [Codespaces](https://github.com/nf-core/dmscore/codespaces) +- Open the repo in [Codespaces](https://github.com/nf-core/deepmutscan/codespaces) - Tools installed - nf-core - Nextflow diff --git a/.github/ISSUE_TEMPLATE/bug_report.yml b/.github/ISSUE_TEMPLATE/bug_report.yml index 58a5523..78f6876 100644 --- a/.github/ISSUE_TEMPLATE/bug_report.yml +++ b/.github/ISSUE_TEMPLATE/bug_report.yml @@ -8,7 +8,7 @@ body: Before you post this issue, please check the documentation: - [nf-core website: troubleshooting](https://nf-co.re/usage/troubleshooting) - - [nf-core/dmscore pipeline documentation](https://nf-co.re/dmscore/usage) + - [nf-core/deepmutscan pipeline documentation](https://nf-co.re/deepmutscan/usage) - type: textarea id: description attributes: @@ -46,4 +46,4 @@ body: * Executor _(eg. slurm, local, awsbatch)_ * Container engine: _(e.g. Docker, Singularity, Conda, Podman, Shifter, Charliecloud, or Apptainer)_ * OS _(eg. CentOS Linux, macOS, Linux Mint)_ - * Version of nf-core/dmscore _(eg. 1.1, 1.5, 1.8.2)_ + * Version of nf-core/deepmutscan _(eg. 1.1, 1.5, 1.8.2)_ diff --git a/.github/ISSUE_TEMPLATE/config.yml b/.github/ISSUE_TEMPLATE/config.yml index 08f0c49..572e0cf 100644 --- a/.github/ISSUE_TEMPLATE/config.yml +++ b/.github/ISSUE_TEMPLATE/config.yml @@ -2,6 +2,6 @@ contact_links: - name: Join nf-core url: https://nf-co.re/join about: Please join the nf-core community here - - name: "Slack #dmscore channel" - url: https://nfcore.slack.com/channels/dmscore - about: Discussion about the nf-core/dmscore pipeline + - name: "Slack #deepmutscan channel" + url: https://nfcore.slack.com/channels/deepmutscan + about: Discussion about the nf-core/deepmutscan pipeline diff --git a/.github/ISSUE_TEMPLATE/feature_request.yml b/.github/ISSUE_TEMPLATE/feature_request.yml index a08dd5f..0731019 100644 --- a/.github/ISSUE_TEMPLATE/feature_request.yml +++ b/.github/ISSUE_TEMPLATE/feature_request.yml @@ -1,5 +1,5 @@ name: Feature request -description: Suggest an idea for the nf-core/dmscore pipeline +description: Suggest an idea for the nf-core/deepmutscan pipeline labels: enhancement body: - type: textarea diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index 0d6f133..ee9e558 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -1,22 +1,22 @@ ## PR checklist - [ ] This comment contains a description of changes (with reason). - [ ] If you've fixed a bug or added code that should be tested, add tests! -- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/dmscore/tree/master/.github/CONTRIBUTING.md) -- [ ] If necessary, also make a PR on the nf-core/dmscore _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository. +- [ ] If you've added a new tool - have you followed the pipeline conventions in the [contribution docs](https://github.com/nf-core/deepmutscan/tree/master/.github/CONTRIBUTING.md) +- [ ] If necessary, also make a PR on the nf-core/deepmutscan _branch_ on the [nf-core/test-datasets](https://github.com/nf-core/test-datasets) repository. - [ ] Make sure your code lints (`nf-core pipelines lint`). - [ ] Ensure the test suite passes (`nextflow run . -profile test,docker --outdir `). - [ ] Check for unexpected warnings in debug mode (`nextflow run . -profile debug,test,docker --outdir `). diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml index 50287a8..e47925c 100644 --- a/.github/workflows/awsfulltest.yml +++ b/.github/workflows/awsfulltest.yml @@ -16,7 +16,7 @@ jobs: run-platform: name: Run AWS full tests # run only if the PR is approved by at least 2 reviewers and against the master branch or manually triggered - if: github.repository == 'nf-core/dmscore' && github.event.review.state == 'approved' && github.event.pull_request.base.ref == 'master' || github.event_name == 'workflow_dispatch' + if: github.repository == 'nf-core/deepmutscan' && github.event.review.state == 'approved' && github.event.pull_request.base.ref == 'master' || github.event_name == 'workflow_dispatch' runs-on: ubuntu-latest steps: - name: Get PR reviews @@ -53,11 +53,11 @@ jobs: access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} revision: ${{ github.sha }} - workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/dmscore/work-${{ github.sha }} + workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/deepmutscan/work-${{ github.sha }} parameters: | { "hook_url": "${{ secrets.MEGATESTS_ALERTS_SLACK_HOOK_URL }}", - "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/dmscore/results-${{ github.sha }}" + "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/deepmutscan/results-${{ github.sha }}" } profiles: test_full diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 4a4b5db..0f92edd 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -7,7 +7,7 @@ on: jobs: run-platform: name: Run AWS tests - if: github.repository == 'nf-core/dmscore' + if: github.repository == 'nf-core/deepmutscan' runs-on: ubuntu-latest steps: # Launch workflow using Seqera Platform CLI tool action @@ -18,10 +18,10 @@ jobs: access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} revision: ${{ github.sha }} - workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/dmscore/work-${{ github.sha }} + workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/deepmutscan/work-${{ github.sha }} parameters: | { - "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/dmscore/results-test-${{ github.sha }}" + "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/deepmutscan/results-test-${{ github.sha }}" } profiles: test diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index 92e3db0..d13cf80 100644 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -13,9 +13,9 @@ jobs: steps: # PRs to the nf-core repo main/master branch are only ok if coming from the nf-core repo `dev` or any `patch` branches - name: Check PRs - if: github.repository == 'nf-core/dmscore' + if: github.repository == 'nf-core/deepmutscan' run: | - { [[ ${{github.event.pull_request.head.repo.full_name }} == nf-core/dmscore ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] + { [[ ${{github.event.pull_request.head.repo.full_name }} == nf-core/deepmutscan ]] && [[ $GITHUB_HEAD_REF == "dev" ]]; } || [[ $GITHUB_HEAD_REF == "patch" ]] # If the above check failed, post a comment on the PR explaining the failure # NOTE - this doesn't currently work if the PR is coming from a fork, due to limitations in GitHub actions secrets diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 92396ce..c23beda 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -22,7 +22,7 @@ jobs: test: name: "Run pipeline with test data (${{ matrix.NXF_VER }} | ${{ matrix.test_name }} | ${{ matrix.profile }})" # Only run on push if this is the nf-core dev branch (merged PRs) - if: "${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/dmscore') }}" + if: "${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/deepmutscan') }}" runs-on: ubuntu-latest strategy: matrix: diff --git a/.github/workflows/fix-linting.yml b/.github/workflows/fix-linting.yml index addd34e..8bda64e 100644 --- a/.github/workflows/fix-linting.yml +++ b/.github/workflows/fix-linting.yml @@ -9,7 +9,7 @@ jobs: if: > contains(github.event.comment.html_url, '/pull/') && contains(github.event.comment.body, '@nf-core-bot fix linting') && - github.repository == 'nf-core/dmscore' + github.repository == 'nf-core/deepmutscan' runs-on: ubuntu-latest steps: # Use the @nf-core-bot token to check out so we can push later @@ -86,4 +86,4 @@ jobs: issue-number: ${{ github.event.issue.number }} body: | @${{ github.actor }} I tried to fix the linting errors, but it didn't work. Please fix them manually. - See [CI log](https://github.com/nf-core/dmscore/actions/runs/${{ github.run_id }}) for more details. + See [CI log](https://github.com/nf-core/deepmutscan/actions/runs/${{ github.run_id }}) for more details. diff --git a/.nf-core.yml b/.nf-core.yml index 59e948b..4ecfff4 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -1,12 +1,8 @@ -repository_type: pipeline - -nf_core_version: 3.1.2 - lint: {} - +nf_core_version: 3.1.2 +repository_type: pipeline template: - org: nf-core - name: dmscore + author: Benjamin Wehnert & Max Stammnitz description: "Until now, most Deep Mutational Scanning (DMS) experiments relied\ \ on variant-specific barcoded libraries for sequencing. This method enabled DMS\ \ on large proteins and led to many great publications. Recently, efforts have\ @@ -17,8 +13,9 @@ template: \ files and generating a count table of variants. Along the way, it provides multiple\ \ QC metrics, enabling users to quickly evaluate the success of their experimental\ \ setup." - author: Benjamin Wehnert & Max Stammnitz - version: 1.0.0dev - force: true - outdir: . + force: false is_nfcore: true + name: deepmutscan + org: nf-core + outdir: . + version: 1.0.0dev diff --git a/CHANGELOG.md b/CHANGELOG.md index 40ea82f..8d3f816 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -1,11 +1,11 @@ -# nf-core/dmscore: Changelog +# nf-core/deepmutscan: Changelog The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html). ## v1.0.0dev - [date] -Initial release of nf-core/dmscore, created with the [nf-core](https://nf-co.re/) template. +Initial release of nf-core/deepmutscan, created with the [nf-core](https://nf-co.re/) template. ### `Added` diff --git a/CITATIONS.md b/CITATIONS.md index a7e69f7..3ea1015 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -1,4 +1,4 @@ -# nf-core/dmscore: Citations +# nf-core/deepmutscan: Citations ## [nf-core](https://pubmed.ncbi.nlm.nih.gov/32055031/) diff --git a/LICENSE b/LICENSE index 48133aa..8119d7b 100644 --- a/LICENSE +++ b/LICENSE @@ -1,6 +1,6 @@ MIT License -Copyright (c) The nf-core/dmscore team +Copyright (c) The nf-core/deepmutscan team Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation files (the "Software"), to deal diff --git a/README.md b/README.md index b6ae647..3898d62 100644 --- a/README.md +++ b/README.md @@ -1,25 +1,25 @@

- - nf-core/dmscore + + nf-core/deepmutscan

-[![GitHub Actions CI Status](https://github.com/nf-core/dmscore/actions/workflows/ci.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/ci.yml) -[![GitHub Actions Linting Status](https://github.com/nf-core/dmscore/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/dmscore/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) +[![GitHub Actions CI Status](https://github.com/nf-core/deepmutscan/actions/workflows/ci.yml/badge.svg)](https://github.com/nf-core/deepmutscan/actions/workflows/ci.yml) +[![GitHub Actions Linting Status](https://github.com/nf-core/deepmutscan/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/deepmutscan/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/deepmutscan/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) [![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com) [![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A524.04.2-23aa62.svg)](https://www.nextflow.io/) [![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/) [![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/) [![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/) -[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/dmscore) +[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/deepmutscan) -[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23dmscore-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/dmscore)[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core) +[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23deepmutscan-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/deepmutscan)[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core) ## Introduction -**nf-core/dmscore** is a bioinformatics pipeline that ... +**nf-core/deepmutscan** is a bioinformatics pipeline that ... ```bash -nextflow run nf-core/dmscore \ +nextflow run nf-core/deepmutscan \ -profile \ --input samplesheet.csv \ --outdir @@ -66,17 +66,17 @@ nextflow run nf-core/dmscore \ > [!WARNING] > Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/usage/getting_started/configuration#custom-configuration-files). -For more details and further functionality, please refer to the [usage documentation](https://nf-co.re/dmscore/usage) and the [parameter documentation](https://nf-co.re/dmscore/parameters). +For more details and further functionality, please refer to the [usage documentation](https://nf-co.re/deepmutscan/usage) and the [parameter documentation](https://nf-co.re/deepmutscan/parameters). ## Pipeline output -To see the results of an example test run with a full size dataset refer to the [results](https://nf-co.re/dmscore/results) tab on the nf-core website pipeline page. +To see the results of an example test run with a full size dataset refer to the [results](https://nf-co.re/deepmutscan/results) tab on the nf-core website pipeline page. For more details about the output files and reports, please refer to the -[output documentation](https://nf-co.re/dmscore/output). +[output documentation](https://nf-co.re/deepmutscan/output). ## Credits -nf-core/dmscore was originally written by Benjamin Wehnert & Max Stammnitz. +nf-core/deepmutscan was originally written by Benjamin Wehnert & Max Stammnitz. We thank the following people for their extensive assistance in the development of this pipeline: @@ -86,12 +86,12 @@ We thank the following people for their extensive assistance in the development If you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md). -For further information or help, don't hesitate to get in touch on the [Slack `#dmscore` channel](https://nfcore.slack.com/channels/dmscore) (you can join with [this invite](https://nf-co.re/join/slack)). +For further information or help, don't hesitate to get in touch on the [Slack `#deepmutscan` channel](https://nfcore.slack.com/channels/deepmutscan) (you can join with [this invite](https://nf-co.re/join/slack)). ## Citations - + diff --git a/assets/adaptivecard.json b/assets/adaptivecard.json index 62ad62a..90786c9 100644 --- a/assets/adaptivecard.json +++ b/assets/adaptivecard.json @@ -17,7 +17,7 @@ "size": "Large", "weight": "Bolder", "color": "<% if (success) { %>Good<% } else { %>Attention<%} %>", - "text": "nf-core/dmscore v${version} - ${runName}", + "text": "nf-core/deepmutscan v${version} - ${runName}", "wrap": true }, { diff --git a/assets/email_template.html b/assets/email_template.html index 8baab9e..c051109 100644 --- a/assets/email_template.html +++ b/assets/email_template.html @@ -4,21 +4,21 @@ - - nf-core/dmscore Pipeline Report + + nf-core/deepmutscan Pipeline Report
-

nf-core/dmscore ${version}

+

nf-core/deepmutscan ${version}

Run Name: $runName

<% if (!success){ out << """
diff --git a/assets/email_template.txt b/assets/email_template.txt index 4247bc4..907c089 100644 --- a/assets/email_template.txt +++ b/assets/email_template.txt @@ -4,15 +4,15 @@ |\\ | |__ __ / ` / \\ |__) |__ } { | \\| | \\__, \\__/ | \\ |___ \\`-._,-`-, `._,._,' - nf-core/dmscore ${version} + nf-core/deepmutscan ${version} ---------------------------------------------------- Run Name: $runName <% if (success){ - out << "## nf-core/dmscore execution completed successfully! ##" + out << "## nf-core/deepmutscan execution completed successfully! ##" } else { out << """#################################################### -## nf-core/dmscore execution completed unsuccessfully! ## +## nf-core/deepmutscan execution completed unsuccessfully! ## #################################################### The exit status of the task that caused the workflow execution to fail was: $exitStatus. The full error message was: @@ -35,5 +35,5 @@ Pipeline Configuration: <% out << summary.collect{ k,v -> " - $k: $v" }.join("\n") %> -- -nf-core/dmscore -https://github.com/nf-core/dmscore +nf-core/deepmutscan +https://github.com/nf-core/deepmutscan diff --git a/assets/methods_description_template.yml b/assets/methods_description_template.yml index e4e7ab3..0076d67 100644 --- a/assets/methods_description_template.yml +++ b/assets/methods_description_template.yml @@ -1,13 +1,13 @@ -id: "nf-core-dmscore-methods-description" +id: "nf-core-deepmutscan-methods-description" description: "Suggested text and references to use when describing pipeline usage within the methods section of a publication." -section_name: "nf-core/dmscore Methods Description" -section_href: "https://github.com/nf-core/dmscore" +section_name: "nf-core/deepmutscan Methods Description" +section_href: "https://github.com/nf-core/deepmutscan" plot_type: "html" ## TODO nf-core: Update the HTML below to your preferred methods description, e.g. add publication citation for this pipeline ## You inject any metadata in the Nextflow '${workflow}' object data: |

Methods

-

Data was processed using nf-core/dmscore v${workflow.manifest.version} ${doi_text} of the nf-core collection of workflows (Ewels et al., 2020), utilising reproducible software environments from the Bioconda (Grüning et al., 2018) and Biocontainers (da Veiga Leprevost et al., 2017) projects.

+

Data was processed using nf-core/deepmutscan v${workflow.manifest.version} ${doi_text} of the nf-core collection of workflows (Ewels et al., 2020), utilising reproducible software environments from the Bioconda (Grüning et al., 2018) and Biocontainers (da Veiga Leprevost et al., 2017) projects.

The pipeline was executed with Nextflow v${workflow.nextflow.version} (Di Tommaso et al., 2017) with the following command:

${workflow.commandLine}

${tool_citations}

diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 2f6ceb9..c27516b 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -1,13 +1,13 @@ report_comment: > - This report has been generated by the nf-core/dmscore + This report has been generated by the nf-core/deepmutscan analysis pipeline. 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Assumes that all software is installed and available on diff --git a/conf/test.config b/conf/test.config index 8c566af..ae518f2 100644 --- a/conf/test.config +++ b/conf/test.config @@ -5,7 +5,7 @@ Defines input files and everything required to run a fast and simple pipeline test. Use as follows: - nextflow run nf-core/dmscore -profile test, --outdir + nextflow run nf-core/deepmutscan -profile test, --outdir ---------------------------------------------------------------------------------------- */ diff --git a/conf/test_full.config b/conf/test_full.config index b57a75c..27ac392 100644 --- a/conf/test_full.config +++ b/conf/test_full.config @@ -5,7 +5,7 @@ Defines input files and everything required to run a full size pipeline test. Use as follows: - nextflow run nf-core/dmscore -profile test_full, --outdir + nextflow run nf-core/deepmutscan -profile test_full, --outdir ---------------------------------------------------------------------------------------- */ diff --git a/docs/README.md b/docs/README.md index 917d984..657c1b3 100644 --- a/docs/README.md +++ b/docs/README.md @@ -1,6 +1,6 @@ -# nf-core/dmscore: Documentation +# nf-core/deepmutscan: Documentation -The nf-core/dmscore documentation is split into the following pages: +The nf-core/deepmutscan documentation is split into the following pages: - [Usage](usage.md) - An overview of how the pipeline works, how to run it and a description of all of the different command-line flags. diff --git a/docs/images/nf-core-deepmutscan_logo_dark.png b/docs/images/nf-core-deepmutscan_logo_dark.png new file mode 100644 index 0000000000000000000000000000000000000000..8b387faba425d60bebf7bafaf89552133aa33485 GIT binary patch literal 28299 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Usage +# nf-core/deepmutscan: Usage -## :warning: Please read this documentation on the nf-core website: [https://nf-co.re/dmscore/usage](https://nf-co.re/dmscore/usage) +## :warning: Please read this documentation on the nf-core website: [https://nf-co.re/deepmutscan/usage](https://nf-co.re/deepmutscan/usage) > _Documentation of pipeline parameters is generated automatically from the pipeline schema and can no longer be found in markdown files._ @@ -57,7 +57,7 @@ An [example samplesheet](../assets/samplesheet.csv) has been provided with the p The typical command for running the pipeline is as follows: ```bash -nextflow run nf-core/dmscore --input ./samplesheet.csv --outdir ./results --genome GRCh37 -profile docker +nextflow run nf-core/deepmutscan --input ./samplesheet.csv --outdir ./results --genome GRCh37 -profile docker ``` This will launch the pipeline with the `docker` configuration profile. See below for more information about profiles. @@ -81,7 +81,7 @@ Pipeline settings can be provided in a `yaml` or `json` file via `-params-file < The above pipeline run specified with a params file in yaml format: ```bash -nextflow run nf-core/dmscore -profile docker -params-file params.yaml +nextflow run nf-core/deepmutscan -profile docker -params-file params.yaml ``` with: @@ -100,14 +100,14 @@ You can also generate such `YAML`/`JSON` files via [nf-core/launch](https://nf-c When you run the above command, Nextflow automatically pulls the pipeline code from GitHub and stores it as a cached version. When running the pipeline after this, it will always use the cached version if available - even if the pipeline has been updated since. To make sure that you're running the latest version of the pipeline, make sure that you regularly update the cached version of the pipeline: ```bash -nextflow pull nf-core/dmscore +nextflow pull nf-core/deepmutscan ``` ### Reproducibility It is a good idea to specify the pipeline version when running the pipeline on your data. This ensures that a specific version of the pipeline code and software are used when you run your pipeline. If you keep using the same tag, you'll be running the same version of the pipeline, even if there have been changes to the code since. -First, go to the [nf-core/dmscore releases page](https://github.com/nf-core/dmscore/releases) and find the latest pipeline version - numeric only (eg. `1.3.1`). Then specify this when running the pipeline with `-r` (one hyphen) - eg. `-r 1.3.1`. Of course, you can switch to another version by changing the number after the `-r` flag. +First, go to the [nf-core/deepmutscan releases page](https://github.com/nf-core/deepmutscan/releases) and find the latest pipeline version - numeric only (eg. `1.3.1`). Then specify this when running the pipeline with `-r` (one hyphen) - eg. `-r 1.3.1`. Of course, you can switch to another version by changing the number after the `-r` flag. This version number will be logged in reports when you run the pipeline, so that you'll know what you used when you look back in the future. For example, at the bottom of the MultiQC reports. diff --git a/main.nf b/main.nf index 8c9943a..d668cea 100644 --- a/main.nf +++ b/main.nf @@ -1,11 +1,11 @@ #!/usr/bin/env nextflow /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - nf-core/dmscore + nf-core/deepmutscan ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - Github : https://github.com/nf-core/dmscore - Website: https://nf-co.re/dmscore - Slack : https://nfcore.slack.com/channels/dmscore + Github : https://github.com/nf-core/deepmutscan + Website: https://nf-co.re/deepmutscan + Slack : https://nfcore.slack.com/channels/deepmutscan ---------------------------------------------------------------------------------------- */ @@ -15,10 +15,10 @@ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -include { DMSCORE } from './workflows/dmscore' -include { PIPELINE_INITIALISATION } from './subworkflows/local/utils_nfcore_dmscore_pipeline' -include { PIPELINE_COMPLETION } from './subworkflows/local/utils_nfcore_dmscore_pipeline' -include { getGenomeAttribute } from './subworkflows/local/utils_nfcore_dmscore_pipeline' +include { DEEPMUTSCAN } from './workflows/deepmutscan' +include { PIPELINE_INITIALISATION } from './subworkflows/local/utils_nfcore_deepmutscan_pipeline' +include { PIPELINE_COMPLETION } from './subworkflows/local/utils_nfcore_deepmutscan_pipeline' +include { getGenomeAttribute } from './subworkflows/local/utils_nfcore_deepmutscan_pipeline' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -40,7 +40,7 @@ params.fasta = getGenomeAttribute('fasta') // // WORKFLOW: Run main analysis pipeline depending on type of input // -workflow NFCORE_DMSCORE { +workflow NFCORE_DEEPMUTSCAN { take: samplesheet // channel: samplesheet read in from --input @@ -50,11 +50,11 @@ workflow NFCORE_DMSCORE { // // WORKFLOW: Run pipeline // - DMSCORE ( + DEEPMUTSCAN ( samplesheet ) emit: - multiqc_report = DMSCORE.out.multiqc_report // channel: /path/to/multiqc_report.html + multiqc_report = DEEPMUTSCAN.out.multiqc_report // channel: /path/to/multiqc_report.html } /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -80,7 +80,7 @@ workflow { // // WORKFLOW: Run main workflow // - NFCORE_DMSCORE ( + NFCORE_DEEPMUTSCAN ( PIPELINE_INITIALISATION.out.samplesheet ) // @@ -93,7 +93,7 @@ workflow { params.outdir, params.monochrome_logs, params.hook_url, - NFCORE_DMSCORE.out.multiqc_report + NFCORE_DEEPMUTSCAN.out.multiqc_report ) } diff --git a/modules.json b/modules.json index 910602b..e30d1fa 100644 --- a/modules.json +++ b/modules.json @@ -1,6 +1,6 @@ { - "name": "nf-core/dmscore", - "homePage": "https://github.com/nf-core/dmscore", + "name": "nf-core/deepmutscan", + "homePage": "https://github.com/nf-core/deepmutscan", "repos": { "https://github.com/nf-core/modules.git": { "modules": { diff --git a/nextflow.config b/nextflow.config index 3fd3e96..bb62d02 100644 --- a/nextflow.config +++ b/nextflow.config @@ -1,6 +1,6 @@ /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - nf-core/dmscore Nextflow config file + nf-core/deepmutscan Nextflow config file ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Default config options for all compute environments ---------------------------------------------------------------------------------------- @@ -167,9 +167,9 @@ profiles { // Load nf-core custom profiles from different Institutions includeConfig !System.getenv('NXF_OFFLINE') && params.custom_config_base ? "${params.custom_config_base}/nfcore_custom.config" : "/dev/null" -// Load nf-core/dmscore custom profiles from different institutions. +// Load nf-core/deepmutscan custom profiles from different institutions. // TODO nf-core: Optionally, you can add a pipeline-specific nf-core config at https://github.com/nf-core/configs -// includeConfig !System.getenv('NXF_OFFLINE') && params.custom_config_base ? "${params.custom_config_base}/pipeline/dmscore.config" : "/dev/null" +// includeConfig !System.getenv('NXF_OFFLINE') && params.custom_config_base ? "${params.custom_config_base}/pipeline/deepmutscan.config" : "/dev/null" // Set default registry for Apptainer, Docker, Podman, Charliecloud and Singularity independent of -profile // Will not be used unless Apptainer / Docker / Podman / Charliecloud / Singularity are enabled @@ -225,7 +225,7 @@ dag { } manifest { - name = 'nf-core/dmscore' + name = 'nf-core/deepmutscan' author = """Benjamin Wehnert & Max Stammnitz""" // The author field is deprecated from Nextflow version 24.10.0, use contributors instead contributors = [ // TODO nf-core: Update the field with the details of the contributors to your pipeline. New with Nextflow version 24.10.0 @@ -238,7 +238,7 @@ manifest { orcid: '' ], ] - homePage = 'https://github.com/nf-core/dmscore' + homePage = 'https://github.com/nf-core/deepmutscan' description = """Until now, most Deep Mutational Scanning (DMS) experiments relied on variant-specific barcoded libraries for sequencing. This method enabled DMS on large proteins and led to many great publications. Recently, efforts have increased to make use of the classic and more simple random fragmentation-based short-read sequencing (“shotgun-sequencing”). This saves time and money and due to its simpler experimental design is less prone to mistakes. dmscore handles the essential computational steps, processing the raw FASTQ files and generating a count table of variants. Along the way, it provides multiple QC metrics, enabling users to quickly evaluate the success of their experimental setup.""" mainScript = 'main.nf' defaultBranch = 'master' @@ -257,7 +257,7 @@ validation { monochromeLogs = params.monochrome_logs help { enabled = true - command = "nextflow run nf-core/dmscore -profile --input samplesheet.csv --outdir " + command = "nextflow run nf-core/deepmutscan -profile --input samplesheet.csv --outdir " fullParameter = "help_full" showHiddenParameter = "show_hidden" beforeText = """ @@ -267,7 +267,7 @@ validation { \033[0;34m |\\ | |__ __ / ` / \\ |__) |__ \033[0;33m} {\033[0m \033[0;34m | \\| | \\__, \\__/ | \\ |___ \033[0;32m\\`-._,-`-,\033[0m \033[0;32m`._,._,\'\033[0m -\033[0;35m nf-core/dmscore ${manifest.version}\033[0m +\033[0;35m nf-core/deepmutscan ${manifest.version}\033[0m -\033[2m----------------------------------------------------\033[0m- """ afterText = """${manifest.doi ? "\n* The pipeline\n" : ""}${manifest.doi.tokenize(",").collect { " https://doi.org/${it.trim().replace('https://doi.org/','')}"}.join("\n")}${manifest.doi ? "\n" : ""} @@ -275,7 +275,7 @@ validation { https://doi.org/10.1038/s41587-020-0439-x * Software dependencies - https://github.com/nf-core/dmscore/blob/master/CITATIONS.md + https://github.com/nf-core/deepmutscan/blob/master/CITATIONS.md """ } summary { diff --git a/nextflow_schema.json b/nextflow_schema.json index e4e5523..735acee 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -1,7 +1,7 @@ { "$schema": "https://json-schema.org/draft/2020-12/schema", - "$id": "https://raw.githubusercontent.com/nf-core/dmscore/master/nextflow_schema.json", - "title": "nf-core/dmscore pipeline parameters", + "$id": "https://raw.githubusercontent.com/nf-core/deepmutscan/master/nextflow_schema.json", + "title": "nf-core/deepmutscan pipeline parameters", "description": "Until now, most Deep Mutational Scanning (DMS) experiments relied on variant-specific barcoded libraries for sequencing. This method enabled DMS on large proteins and led to many great publications. Recently, efforts have increased to make use of the classic and more simple random fragmentation-based short-read sequencing (“shotgun-sequencing”). This saves time and money and due to its simpler experimental design is less prone to mistakes. dmscore handles the essential computational steps, processing the raw FASTQ files and generating a count table of variants. Along the way, it provides multiple QC metrics, enabling users to quickly evaluate the success of their experimental setup.", "type": "object", "$defs": { @@ -20,7 +20,7 @@ "mimetype": "text/csv", "pattern": "^\\S+\\.csv$", "description": "Path to comma-separated file containing information about the samples in the experiment.", - "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 3 columns, and a header row. See [usage docs](https://nf-co.re/dmscore/usage#samplesheet-input).", + "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 3 columns, and a header row. See [usage docs](https://nf-co.re/deepmutscan/usage#samplesheet-input).", "fa_icon": "fas fa-file-csv" }, "outdir": { diff --git a/ro-crate-metadata.json b/ro-crate-metadata.json index 766d934..f96d41c 100644 --- a/ro-crate-metadata.json +++ b/ro-crate-metadata.json @@ -22,8 +22,8 @@ "@id": "./", "@type": "Dataset", "creativeWorkStatus": "InProgress", - "datePublished": "2025-01-22T16:52:07+00:00", - "description": "

\n \n \n \"nf-core/dmscore\"\n \n

\n\n[![GitHub Actions CI Status](https://github.com/nf-core/dmscore/actions/workflows/ci.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/ci.yml)\n[![GitHub Actions Linting Status](https://github.com/nf-core/dmscore/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/dmscore/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/dmscore/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)\n[![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com)\n\n[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A524.04.2-23aa62.svg)](https://www.nextflow.io/)\n[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)\n[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)\n[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)\n[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/dmscore)\n\n[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23dmscore-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/dmscore)[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)\n\n## Introduction\n\n**nf-core/dmscore** is a bioinformatics pipeline that ...\n\n\n\n\n1. Read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/))2. Present QC for raw reads ([`MultiQC`](http://multiqc.info/))\n\n## Usage\n\n> [!NOTE]\n> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/usage/installation) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/usage/introduction#how-to-run-a-pipeline) with `-profile test` before running the workflow on actual data.\n\n\n\nNow, you can run the pipeline using:\n\n\n\n```bash\nnextflow run nf-core/dmscore \\\n -profile \\\n --input samplesheet.csv \\\n --outdir \n```\n\n> [!WARNING]\n> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/usage/getting_started/configuration#custom-configuration-files).\n\nFor more details and further functionality, please refer to the [usage documentation](https://nf-co.re/dmscore/usage) and the [parameter documentation](https://nf-co.re/dmscore/parameters).\n\n## Pipeline output\n\nTo see the results of an example test run with a full size dataset refer to the [results](https://nf-co.re/dmscore/results) tab on the nf-core website pipeline page.\nFor more details about the output files and reports, please refer to the\n[output documentation](https://nf-co.re/dmscore/output).\n\n## Credits\n\nnf-core/dmscore was originally written by Benjamin Wehnert & Max Stammnitz.\n\nWe thank the following people for their extensive assistance in the development of this pipeline:\n\n\n\n## Contributions and Support\n\nIf you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md).\n\nFor further information or help, don't hesitate to get in touch on the [Slack `#dmscore` channel](https://nfcore.slack.com/channels/dmscore) (you can join with [this invite](https://nf-co.re/join/slack)).\n\n## Citations\n\n\n\n\n\n\nAn extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.\n\nYou can cite the `nf-core` publication as follows:\n\n> **The nf-core framework for community-curated bioinformatics pipelines.**\n>\n> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.\n>\n> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).\n", + "datePublished": "2025-08-28T15:55:11+00:00", + "description": "

\n \n \n \"nf-core/deepmutscan\"\n \n

\n\n[![GitHub Actions CI Status](https://github.com/nf-core/deepmutscan/actions/workflows/ci.yml/badge.svg)](https://github.com/nf-core/deepmutscan/actions/workflows/ci.yml)\n[![GitHub Actions Linting Status](https://github.com/nf-core/deepmutscan/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/deepmutscan/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/deepmutscan/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)\n[![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com)\n\n[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A524.04.2-23aa62.svg)](https://www.nextflow.io/)\n[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)\n[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)\n[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)\n[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/deepmutscan)\n\n[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23deepmutscan-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/deepmutscan)[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)\n\n## Introduction\n\n**nf-core/deepmutscan** is a bioinformatics pipeline that ...\n\n\n\n\n1. Read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/))2. Present QC for raw reads ([`MultiQC`](http://multiqc.info/))\n\n## Usage\n\n> [!NOTE]\n> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/usage/installation) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/usage/introduction#how-to-run-a-pipeline) with `-profile test` before running the workflow on actual data.\n\n\n\nNow, you can run the pipeline using:\n\n\n\n```bash\nnextflow run nf-core/deepmutscan \\\n -profile \\\n --input samplesheet.csv \\\n --outdir \n```\n\n> [!WARNING]\n> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/usage/getting_started/configuration#custom-configuration-files).\n\nFor more details and further functionality, please refer to the [usage documentation](https://nf-co.re/deepmutscan/usage) and the [parameter documentation](https://nf-co.re/deepmutscan/parameters).\n\n## Pipeline output\n\nTo see the results of an example test run with a full size dataset refer to the [results](https://nf-co.re/deepmutscan/results) tab on the nf-core website pipeline page.\nFor more details about the output files and reports, please refer to the\n[output documentation](https://nf-co.re/deepmutscan/output).\n\n## Credits\n\nnf-core/deepmutscan was originally written by Benjamin Wehnert & Max Stammnitz.\n\nWe thank the following people for their extensive assistance in the development of this pipeline:\n\n\n\n## Contributions and Support\n\nIf you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md).\n\nFor further information or help, don't hesitate to get in touch on the [Slack `#deepmutscan` channel](https://nfcore.slack.com/channels/deepmutscan) (you can join with [this invite](https://nf-co.re/join/slack)).\n\n## Citations\n\n\n\n\n\n\nAn extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.\n\nYou can cite the `nf-core` publication as follows:\n\n> **The nf-core framework for community-curated bioinformatics pipelines.**\n>\n> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.\n>\n> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).\n", "hasPart": [ { "@id": "main.nf" @@ -92,17 +92,17 @@ "@id": ".prettierignore" } ], - "isBasedOn": "https://github.com/nf-core/dmscore", + "isBasedOn": "https://github.com/nf-core/deepmutscan", "license": "MIT", "mainEntity": { "@id": "main.nf" }, "mentions": [ { - "@id": "#f9815654-37bd-4781-a133-ab36324210f5" + "@id": "#4104f8a2-6a7e-4ded-834a-f92a921dceef" } ], - "name": "nf-core/dmscore" + "name": "nf-core/deepmutscan" }, { "@id": "ro-crate-metadata.json", @@ -127,7 +127,7 @@ "ComputationalWorkflow" ], "dateCreated": "", - "dateModified": "2025-01-22T17:52:07Z", + "dateModified": "2025-08-28T17:55:11Z", "dct:conformsTo": "https://bioschemas.org/profiles/ComputationalWorkflow/1.0-RELEASE/", "keywords": [ "nf-core", @@ -137,7 +137,7 @@ "MIT" ], "name": [ - "nf-core/dmscore" + "nf-core/deepmutscan" ], "programmingLanguage": { "@id": "https://w3id.org/workflowhub/workflow-ro-crate#nextflow" @@ -146,8 +146,8 @@ "@id": "https://nf-co.re/" }, "url": [ - "https://github.com/nf-core/dmscore", - "https://nf-co.re/dmscore/dev/" + "https://github.com/nf-core/deepmutscan", + "https://nf-co.re/deepmutscan/dev/" ], "version": [ "1.0.0dev" @@ -166,23 +166,23 @@ "version": "!>=24.04.2" }, { - "@id": "#f9815654-37bd-4781-a133-ab36324210f5", + "@id": "#4104f8a2-6a7e-4ded-834a-f92a921dceef", "@type": "TestSuite", "instance": [ { - "@id": "#34dab5c1-0a1b-41b0-80d1-74e2f2d38434" + "@id": "#07403d27-5fa6-4937-a914-ac033a0072a4" } ], "mainEntity": { "@id": "main.nf" }, - "name": "Test suite for nf-core/dmscore" + "name": "Test suite for nf-core/deepmutscan" }, { - "@id": "#34dab5c1-0a1b-41b0-80d1-74e2f2d38434", + "@id": "#07403d27-5fa6-4937-a914-ac033a0072a4", "@type": "TestInstance", - "name": "GitHub Actions workflow for testing nf-core/dmscore", - "resource": "repos/nf-core/dmscore/actions/workflows/ci.yml", + "name": "GitHub Actions workflow for testing nf-core/deepmutscan", + "resource": "repos/nf-core/deepmutscan/actions/workflows/ci.yml", "runsOn": { "@id": "https://w3id.org/ro/terms/test#GithubService" }, diff --git a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf b/subworkflows/local/utils_nfcore_deepmutscan_pipeline/main.nf similarity index 99% rename from subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf rename to subworkflows/local/utils_nfcore_deepmutscan_pipeline/main.nf index 14eba3c..cb2380d 100644 --- a/subworkflows/local/utils_nfcore_dmscore_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_deepmutscan_pipeline/main.nf @@ -1,5 +1,5 @@ // -// Subworkflow with functionality specific to the nf-core/dmscore pipeline +// Subworkflow with functionality specific to the nf-core/deepmutscan pipeline // /* diff --git a/workflows/dmscore.nf b/workflows/deepmutscan.nf similarity index 96% rename from workflows/dmscore.nf rename to workflows/deepmutscan.nf index 041e503..2b6bf8d 100644 --- a/workflows/dmscore.nf +++ b/workflows/deepmutscan.nf @@ -8,7 +8,7 @@ include { MULTIQC } from '../modules/nf-core/multiqc/main' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' -include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmscore_pipeline' +include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_deepmutscan_pipeline' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -16,7 +16,7 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_dmsc ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -workflow DMSCORE { +workflow DEEPMUTSCAN { take: ch_samplesheet // channel: samplesheet read in from --input @@ -39,7 +39,7 @@ workflow DMSCORE { softwareVersionsToYAML(ch_versions) .collectFile( storeDir: "${params.outdir}/pipeline_info", - name: 'nf_core_' + 'dmscore_software_' + 'mqc_' + 'versions.yml', + name: 'nf_core_' + 'deepmutscan_software_' + 'mqc_' + 'versions.yml', sort: true, newLine: true ).set { ch_collated_versions } From 868cc8fd99d72841baf05a86b3564d6e5a33c071 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Thu, 28 Aug 2025 19:17:07 +0200 Subject: [PATCH 250/317] finishing renaming process --- conf/base.config | 6 ++++++ 1 file changed, 6 insertions(+) diff --git a/conf/base.config b/conf/base.config index e4e941d..a0daf47 100644 --- a/conf/base.config +++ b/conf/base.config @@ -19,6 +19,12 @@ process { maxRetries = 1 maxErrors = '-1' + // Disable 'noclobber' and pre-clean versions.yml so modules can overwrite it + beforeScript = ''' + set +o noclobber + rm -f versions.yml || true + ''' + // Process-specific resource requirements // NOTE - Please try and reuse the labels below as much as possible. // These labels are used and recognised by default in DSL2 files hosted on nf-core/modules. From 0709ceb111f3f68d232f11d8c9c71127e66a97ec Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Thu, 28 Aug 2025 20:01:28 +0200 Subject: [PATCH 251/317] make complete caching (-resume) possible --- workflows/deepmutscan.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index 93e6a0a..2ef670b 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -43,7 +43,7 @@ params.run_seqdepth = params.run_seqdepth ?: false // Define fasta file as channel (e.g. for BWA index) Channel .fromPath(params.fasta, checkIfExists: true) - .map { fasta -> tuple( { id: "ref" }, fasta ) } + .map { fasta -> tuple( [id: 'ref'], fasta ) } .set { ch_fasta } // Define reading_frame as channel (e.g. for gatk function) From 4d9325d68c45d2b10b6299f59f76586de33ac0e2 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 28 Aug 2025 22:44:34 +0200 Subject: [PATCH 252/317] Update README.md --- README.md | 17 +++++++++++++++-- 1 file changed, 15 insertions(+), 2 deletions(-) diff --git a/README.md b/README.md index 2ef8821..90a14e5 100644 --- a/README.md +++ b/README.md @@ -16,6 +16,19 @@ [![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/deepmutscan) [![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23deepmutscan-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/deepmutscan)[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core) +# Table Of Contents + +1. **[Overview](#overview)** +1. **[Features of nf-core/deepmutscan](#features of nf-core/deepmutscan)** +1. **[Installation](#installation)** +1. **[Usage](#usage)** +1. **[Input Data](#input)** +1. **[Output Data](#output)** +1. **[Citation](#citation)** +1. **[License](#license)** +1. **[Contributing](#contributing)** +1. **[Contact](#contact)** + ## 1. Overview **nf-core/deepmutscan** is a reproducible, scalable, and community-curated pipeline for analyzing deep mutational scanning (DMS) data using shotgun DNA sequencing. DMS enables researchers to measure the fitness effects of thousands of gene variants simultaneously, helping to classify disease causing mutants in human and animal populations, to learn fundamental rules of virus evolution, protein architecture, splicing or small-molecule interactions. @@ -98,7 +111,7 @@ More options and advanced configuration: [see vignette](link). For further infor --- -## 5. Input Data +## 5. Input The primary pipeline input is a sample sheet `.csv` file listing: @@ -111,7 +124,7 @@ See [sample CSV](assets/samplesheet.csv) for formatting. --- -## 6. Output Data +## 6. Output After execution, the pipeline creates the following directory structure: From 0230915b34328b12408863c8e1aa297368e84031 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 28 Aug 2025 22:45:44 +0200 Subject: [PATCH 253/317] Update README.md --- README.md | 20 ++++++++++---------- 1 file changed, 10 insertions(+), 10 deletions(-) diff --git a/README.md b/README.md index 90a14e5..5f25f23 100644 --- a/README.md +++ b/README.md @@ -29,7 +29,7 @@ 1. **[Contributing](#contributing)** 1. **[Contact](#contact)** -## 1. Overview +# Overview **nf-core/deepmutscan** is a reproducible, scalable, and community-curated pipeline for analyzing deep mutational scanning (DMS) data using shotgun DNA sequencing. DMS enables researchers to measure the fitness effects of thousands of gene variants simultaneously, helping to classify disease causing mutants in human and animal populations, to learn fundamental rules of virus evolution, protein architecture, splicing or small-molecule interactions. While DNA synthesis and sequencing technologies have advanced substantially, long open reading frame (ORF) targets still present major challenges for DMS studies. Shotgun DNA sequencing can be used to greatly speed up the inference of long ORF mutant fitness landscapes, theoretically at no expense in accuracy. We have designed the **nf-core/deepmutscan** pipeline to unlock the power of shotgun sequencing based DMS studies on long ORFs, to simplify and standardise the complex bioinformatics steps involved in data processing of such experiments – from read alignment to QC reporting and fitness landscape inferences. @@ -38,7 +38,7 @@ While DNA synthesis and sequencing technologies have advanced substantially, lon --- -## 2. Features of nf-core/deepmutscan +# Features of nf-core/deepmutscan - End-to-end analyses of DMS shotgun sequencing data - Modular, three-stage workflow: alignment → QC → error-aware fitness estimation - Integrates with popular statistical tools like [DiMSum](https://github.com/lehner-lab/DiMSum), [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) @@ -51,7 +51,7 @@ For details of the pipeline and potential future expansions, please consider rea --- -## 3. Installation +# Installation **nf-core/deepmutscan** uses [Nextflow](https://nf-co.re/docs/usage/getting_started/installation), which must be installed on your system: ```bash @@ -70,7 +70,7 @@ For more details and further functionality, please refer to the [usage documenta --- -## 4. Usage +# Usage Prepare: - A **sample sheet** CSV to specify input/output labels, replicates, etc. (see [example](assets/samplesheet.csv)) - A **reference FASTA** file for the gene or region of interest @@ -111,7 +111,7 @@ More options and advanced configuration: [see vignette](link). For further infor --- -## 5. Input +# Input The primary pipeline input is a sample sheet `.csv` file listing: @@ -124,7 +124,7 @@ See [sample CSV](assets/samplesheet.csv) for formatting. --- -## 6. Output +# Output After execution, the pipeline creates the following directory structure: @@ -139,7 +139,7 @@ results/ --- -## 7. Citation +# Citation If you use this pipeline in your research, please cite: > 📄 Wehnert et al., _bioRxiv_ preprint (coming soon) @@ -150,7 +150,7 @@ Please also cite the nf-core framework: --- -## 8. License +# License [MIT License](link) @@ -158,7 +158,7 @@ Please also cite the nf-core framework: --- -## 9. Contributing +# Contributing We welcome contributions from the community! @@ -171,7 +171,7 @@ You can also reach out to us via the **nf-core Slack**, by use of the `#dms` cha --- -## 10. Contact +# Contact For detailled scientific or technical questions, feedback and experimental discussions, feel free to contact us directly: From 077bc6bfaff92756f90f14cec0dc56bc576bd6db Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 28 Aug 2025 22:49:28 +0200 Subject: [PATCH 254/317] Update README.md --- README.md | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/README.md b/README.md index 5f25f23..dcb1c1c 100644 --- a/README.md +++ b/README.md @@ -19,11 +19,11 @@ # Table Of Contents 1. **[Overview](#overview)** -1. **[Features of nf-core/deepmutscan](#features of nf-core/deepmutscan)** +1. **[Features](#features)** 1. **[Installation](#installation)** 1. **[Usage](#usage)** -1. **[Input Data](#input)** -1. **[Output Data](#output)** +1. **[Input](#input)** +1. **[Output](#output)** 1. **[Citation](#citation)** 1. **[License](#license)** 1. **[Contributing](#contributing)** @@ -38,7 +38,7 @@ While DNA synthesis and sequencing technologies have advanced substantially, lon --- -# Features of nf-core/deepmutscan +# Features - End-to-end analyses of DMS shotgun sequencing data - Modular, three-stage workflow: alignment → QC → error-aware fitness estimation - Integrates with popular statistical tools like [DiMSum](https://github.com/lehner-lab/DiMSum), [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) From 0fae26e6e3c1686e07ce57148799db027fa730f5 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 28 Aug 2025 22:52:05 +0200 Subject: [PATCH 255/317] Update README.md --- README.md | 33 ++++++++++----------------------- 1 file changed, 10 insertions(+), 23 deletions(-) diff --git a/README.md b/README.md index dcb1c1c..5dece0a 100644 --- a/README.md +++ b/README.md @@ -16,20 +16,7 @@ [![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/deepmutscan) [![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23deepmutscan-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/deepmutscan)[![Follow on Twitter](http://img.shields.io/badge/twitter-%40nf__core-1DA1F2?labelColor=000000&logo=twitter)](https://twitter.com/nf_core)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core) -# Table Of Contents - -1. **[Overview](#overview)** -1. **[Features](#features)** -1. **[Installation](#installation)** -1. **[Usage](#usage)** -1. **[Input](#input)** -1. **[Output](#output)** -1. **[Citation](#citation)** -1. **[License](#license)** -1. **[Contributing](#contributing)** -1. **[Contact](#contact)** - -# Overview +# 1. Overview **nf-core/deepmutscan** is a reproducible, scalable, and community-curated pipeline for analyzing deep mutational scanning (DMS) data using shotgun DNA sequencing. DMS enables researchers to measure the fitness effects of thousands of gene variants simultaneously, helping to classify disease causing mutants in human and animal populations, to learn fundamental rules of virus evolution, protein architecture, splicing or small-molecule interactions. While DNA synthesis and sequencing technologies have advanced substantially, long open reading frame (ORF) targets still present major challenges for DMS studies. Shotgun DNA sequencing can be used to greatly speed up the inference of long ORF mutant fitness landscapes, theoretically at no expense in accuracy. We have designed the **nf-core/deepmutscan** pipeline to unlock the power of shotgun sequencing based DMS studies on long ORFs, to simplify and standardise the complex bioinformatics steps involved in data processing of such experiments – from read alignment to QC reporting and fitness landscape inferences. @@ -38,7 +25,7 @@ While DNA synthesis and sequencing technologies have advanced substantially, lon --- -# Features +# 2. Features of nf-core/deepmutscan - End-to-end analyses of DMS shotgun sequencing data - Modular, three-stage workflow: alignment → QC → error-aware fitness estimation - Integrates with popular statistical tools like [DiMSum](https://github.com/lehner-lab/DiMSum), [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) @@ -51,7 +38,7 @@ For details of the pipeline and potential future expansions, please consider rea --- -# Installation +# 3. Installation **nf-core/deepmutscan** uses [Nextflow](https://nf-co.re/docs/usage/getting_started/installation), which must be installed on your system: ```bash @@ -70,7 +57,7 @@ For more details and further functionality, please refer to the [usage documenta --- -# Usage +# 4. Usage Prepare: - A **sample sheet** CSV to specify input/output labels, replicates, etc. (see [example](assets/samplesheet.csv)) - A **reference FASTA** file for the gene or region of interest @@ -111,7 +98,7 @@ More options and advanced configuration: [see vignette](link). For further infor --- -# Input +# 5. Input Data The primary pipeline input is a sample sheet `.csv` file listing: @@ -124,7 +111,7 @@ See [sample CSV](assets/samplesheet.csv) for formatting. --- -# Output +# 6. Outputs After execution, the pipeline creates the following directory structure: @@ -139,7 +126,7 @@ results/ --- -# Citation +# 7. Citation If you use this pipeline in your research, please cite: > 📄 Wehnert et al., _bioRxiv_ preprint (coming soon) @@ -150,7 +137,7 @@ Please also cite the nf-core framework: --- -# License +# 8. License [MIT License](link) @@ -158,7 +145,7 @@ Please also cite the nf-core framework: --- -# Contributing +# 9. Contributing We welcome contributions from the community! @@ -171,7 +158,7 @@ You can also reach out to us via the **nf-core Slack**, by use of the `#dms` cha --- -# Contact +# 10. Contact For detailled scientific or technical questions, feedback and experimental discussions, feel free to contact us directly: From 2fc4a3440c9d7e42853aa799acb8eb7bf8475e4b Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Wed, 3 Sep 2025 18:48:39 +0200 Subject: [PATCH 256/317] Update pipeline_steps.md --- docs/pipeline_steps.md | 7 +++++-- 1 file changed, 5 insertions(+), 2 deletions(-) diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md index 568b54e..6b67e99 100644 --- a/docs/pipeline_steps.md +++ b/docs/pipeline_steps.md @@ -81,11 +81,14 @@ This module will implement strategies to distinguish true single nucleotide vari ## 8. Fitness Estimation *(in development)* -The final step of the pipeline will perform fitness estimation based on mutation counts. Planned features include: -- Integration of multiple fitness inference tools beyond DiMSum, including [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) +Using DiMSum, the final step of the pipeline will perform fitness estimation based on mutation counts. Planned features include: +- Integration of multiple other fitness inference tools, including [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) - Options for replicates and condition-specific comparisons - Standardised output formats for downstream analysis and comparison +> [!IMPORTANT] +> We note that exact wildtype sequence reads are filtered out in stage 2. Including synonymous wildtype codons in the original mutagenesis design is therefore essential when it comes to calibrating the fitness calculations. + ------------------------------------------------------------------------ ## Notes for Developers From e1dd3425861a7fbd87278365f0a86668dd88f52d Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Tue, 9 Sep 2025 13:52:55 +0200 Subject: [PATCH 257/317] merging counts for DiMSum input --- conf/modules.config | 8 ++ main.nf | 1 + modules/local/dimsum/merge_counts.nf | 36 +++++++ modules/local/dmsanalysis/bin/merge_counts.R | 100 +++++++++++++++++++ nextflow_schema.json | 5 + workflows/deepmutscan.nf | 41 ++++++++ 6 files changed, 191 insertions(+) create mode 100644 modules/local/dimsum/merge_counts.nf create mode 100644 modules/local/dmsanalysis/bin/merge_counts.R diff --git a/conf/modules.config b/conf/modules.config index 132ccb7..e501251 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -117,6 +117,14 @@ process { } withName: 'GATK_GATKTODIMSUM' { + publishDir = [ + path: "${params.outdir}/DiMSum/single_counts", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + + withName: 'MERGE_COUNTS' { publishDir = [ path: "${params.outdir}/DiMSum", mode: 'copy', diff --git a/main.nf b/main.nf index d668cea..bcee0db 100644 --- a/main.nf +++ b/main.nf @@ -56,6 +56,7 @@ workflow NFCORE_DEEPMUTSCAN { emit: multiqc_report = DEEPMUTSCAN.out.multiqc_report // channel: /path/to/multiqc_report.html } + /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ RUN MAIN WORKFLOW diff --git a/modules/local/dimsum/merge_counts.nf b/modules/local/dimsum/merge_counts.nf new file mode 100644 index 0000000..9d01526 --- /dev/null +++ b/modules/local/dimsum/merge_counts.nf @@ -0,0 +1,36 @@ +process MERGE_COUNTS { + tag "${sample.sample}" + label 'process_single' + publishDir "${params.outdir}/dimsum/${sample.sample}", mode: 'copy' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" + + input: + tuple val(sample), val(metas), path(input_counts), path(output_counts) + path merge_script + + output: + path "counts_merged.tsv", emit: merged_counts + path "versions.yml", emit: versions + + script: + def in_list = input_counts .collect { it.getName() }.join(' ') + def out_list = output_counts.collect { it.getName() }.join(' ') + """ + set -euo pipefail + + Rscript "${merge_script}" \\ + --inputs ${in_list} \\ + --outputs ${out_list} \\ + --out counts_merged.tsv + + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + cat <<-END_VERSIONS > versions.yml + GATK_GATKTODIMSUM: + r-base: \$R_VERSION + END_VERSIONS + """ +} diff --git a/modules/local/dmsanalysis/bin/merge_counts.R b/modules/local/dmsanalysis/bin/merge_counts.R new file mode 100644 index 0000000..db8d3d0 --- /dev/null +++ b/modules/local/dmsanalysis/bin/merge_counts.R @@ -0,0 +1,100 @@ +# merge_counts.R +# Merges DiMSum-ready count tables into a single matrix with columns: +# nt_seq, input1..inputN, output1..outputM + +suppressMessages({ + # keep base R; if you prefer data.table, we can swap in later +}) + +# ---- Core function ---- +merge_dimsum_counts <- function(input_paths, output_paths, out_path = "counts.tsv") { + # input_paths, output_paths: character vectors of file paths + # out_path: output TSV path + + # Helper to read a 2-col TSV without header -> data.frame(nt_seq, count) + read_counts <- function(fp) { + df <- utils::read.table( + fp, header = FALSE, sep = "\t", quote = "", + col.names = c("nt_seq", "count"), + colClasses = c("character", "numeric"), + comment.char = "", check.names = FALSE + ) + df + } + + # Read all inputs / outputs + input_list <- lapply(input_paths, read_counts) + output_list <- lapply(output_paths, read_counts) + + # Collect universe of sequences + all_seqs <- unique(c( + unlist(lapply(input_list, function(x) x$nt_seq)), + unlist(lapply(output_list, function(x) x$nt_seq)) + )) + + # Pre-allocate output frame + n_in <- length(input_list) + n_out <- length(output_list) + col_names <- c( + "nt_seq", + if (n_in > 0) paste0("input", seq_len(n_in)) else character(0), + if (n_out > 0) paste0("output", seq_len(n_out)) else character(0) + ) + # initialize numeric with 0 counts + out <- data.frame( + nt_seq = all_seqs, + matrix(0, nrow = length(all_seqs), ncol = n_in + n_out), + stringsAsFactors = FALSE, check.names = FALSE + ) + names(out) <- col_names + + # Fill inputs + if (n_in > 0) { + for (i in seq_len(n_in)) { + df <- input_list[[i]] + idx <- match(df$nt_seq, out$nt_seq) + out[idx, paste0("input", i)] <- df$count + } + } + + # Fill outputs + if (n_out > 0) { + for (j in seq_len(n_out)) { + df <- output_list[[j]] + idx <- match(df$nt_seq, out$nt_seq) + out[idx, paste0("output", j)] <- df$count + } + } + + # Write TSV with header, no row names, no quotes + utils::write.table(out, file = out_path, sep = "\t", row.names = FALSE, col.names = TRUE, quote = FALSE) + invisible(out) +} + +# ---- Lightweight CLI wrapper ---- +# Usage: +# Rscript merge_counts.R --inputs ... --outputs ... --out counts.tsv +if (sys.nframe() == 0) { + args <- commandArgs(trailingOnly = TRUE) + + # simple flag parser that supports space-separated lists + get_vals <- function(flag) { + if (!(flag %in% args)) return(character(0)) + start <- which(args == flag) + stop <- which(args %in% c("--inputs","--outputs","--out")) + stop <- min(c(stop[stop > start], length(args) + 1)) - 1 + if (stop <= start) return(character(0)) + args[(start + 1):stop] + } + + input_paths <- get_vals("--inputs") + output_paths <- get_vals("--outputs") + out_path <- get_vals("--out") + out_path <- if (length(out_path)) out_path[1] else "counts.tsv" + + if (!length(input_paths) && !length(output_paths)) { + stop("No inputs/outputs provided. Use --inputs and/or --outputs .") + } + + merge_dimsum_counts(input_paths, output_paths, out_path) +} diff --git a/nextflow_schema.json b/nextflow_schema.json index 13bc853..f1fd31e 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -63,6 +63,11 @@ "format": "file-path", "description": "Path to a comma-separated .txt file listing custom codons (e.g., 'AAA,AAC,AAG,...'). Required when mutagenesis_type is 'custom'." }, + "dimsum": { + "type": "boolean", + "default": false, + "description": "Enable DiMSum preparation (group and merge counts per sample into counts_merged.tsv)." + }, "run_seqdepth": { "type": "boolean", "default": false, diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index 2ef670b..67ce1d4 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -20,6 +20,7 @@ include { VISUALIZATION_GLOBAL_POS_BIASES_COV } from '../modules/local/visu include { VISUALIZATION_LOGDIFF } from '../modules/local/visualization/visualization' include { VISUALIZATION_SEQDEPTH } from '../modules/local/visualization/visualization' include { GATK_GATKTODIMSUM } from '../modules/local/gatk/gatktodimsum' +include { MERGE_COUNTS } from '../modules/local/dimsum/merge_counts' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -38,6 +39,7 @@ params.custom_codon_library = params.custom_codon_library ?: '/NULL' params.sliding_window_size = params.sliding_window_size ?: 10 // sliding window size to flatten graphs in plots (e.g. GLOBAL_POS_BIASES_COUNTS function) params.aimed_cov = params.aimed_cov ?: 100 // aimed coverage (assuming equal spread) to visualize threshold in plots params.run_seqdepth = params.run_seqdepth ?: false // creating seqdepth simulation plot, is computationally quite heavy. per default disabled. +params.dimsum = params.dimsum ?: false // run DiMSum for fitness/functionality scores from selection input & output samples // Define fasta file as channel (e.g. for BWA index) @@ -105,6 +107,7 @@ R("modules/local/dmsanalysis/bin/possible_mutations.R").set { possible_mutations R("modules/local/dmsanalysis/bin/prepare_gatk_data_for_count_heatmaps.R").set { prepare_counts_heatmap_script_ch } R("modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R").set { prepare_fitness_heatmap_script_ch } R("modules/local/dmsanalysis/bin/process_raw_gatk.R").set { process_raw_gatk_script_ch } +R("modules/local/dmsanalysis/bin/merge_counts.R").set { merge_counts_script_ch } @@ -365,6 +368,44 @@ logdiff_scriptN = fanoutTo(library_completed_variantCounts_ch, lo ch_script_for_dimsum // path(R script) ) + + +// ----- DiMSum: group per biological sample (from samplesheet) and merge counts to use for DiMSum input ----- + +GATK_GATKTODIMSUM.out.dimsum_input + .map { meta, tsv -> + def s = meta.sample as String + def id = meta.id as String + def base = s ? (s.replaceFirst(/_(input|output|quality)\d+$/, '')) + : (id?.tokenize('_')?.first()) + tuple(base as String, tuple(meta, tsv)) + } + .groupTuple() + .map { base, pairs -> + def metas = pairs.collect { it[0] } + def inputs = pairs.findAll { it[0].type == 'input' }.sort { it[0].replicate }.collect { it[1] } + def outputs = pairs.findAll { it[0].type == 'output' }.sort { it[0].replicate }.collect { it[1] } + tuple([sample: base], metas, inputs, outputs) + } + .filter { smeta, metas, ins, outs -> ins && outs } + .set { ch_dimsum_bundled } + +// Broadcast the singleton script path to match each bundle +def ch_merge_script_for_each = merge_counts_script_ch + .combine(ch_dimsum_bundled) + .map { it[0] } // keep the script path, one per bundle + +// Launch the merge +if (params.dimsum) { + MERGE_COUNTS( + ch_dimsum_bundled, // tuple val(sample), val(metas), path(input_counts), path(output_counts) + ch_merge_script_for_each // path merge_script (broadcast) + ) +} + + + + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From db9c0a9b6cd12ca369c064b141ae397b9c85db3a Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Tue, 9 Sep 2025 15:17:12 +0200 Subject: [PATCH 258/317] experimental design file for dimsum --- conf/modules.config | 16 +++++ .../dimsum/dimsum_experimental_design.nf | 30 ++++++++ .../bin/dimsum_experimentalDesign.R | 69 +++++++++++++++++++ workflows/deepmutscan.nf | 15 ++++ 4 files changed, 130 insertions(+) create mode 100644 modules/local/dimsum/dimsum_experimental_design.nf create mode 100644 modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R diff --git a/conf/modules.config b/conf/modules.config index e501251..de026df 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -131,4 +131,20 @@ process { saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } + + withName: 'MERGE_COUNTS' { + publishDir = [ + path: "${params.outdir}/DiMSum", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + + withName: 'EXPDESIGN_DIMSUM' { + publishDir = [ + path: "${params.outdir}/DiMSum", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } } diff --git a/modules/local/dimsum/dimsum_experimental_design.nf b/modules/local/dimsum/dimsum_experimental_design.nf new file mode 100644 index 0000000..781a768 --- /dev/null +++ b/modules/local/dimsum/dimsum_experimental_design.nf @@ -0,0 +1,30 @@ +process EXPDESIGN_DIMSUM { + tag "experimentalDesign" + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" + + input: + path samplesheet_csv + path script // R file that defines: make_dimsum_experimental_design(input_csv, out_path) + + output: + path "experimentalDesign.tsv", emit: experimental_design + path "versions.yml", emit: versions + + script: + """ + set -euo pipefail + + Rscript -e "source('$script'); make_dimsum_experimental_design('$samplesheet_csv', 'experimentalDesign.tsv')" + + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + cat <<-END_VERSIONS > versions.yml + DIMSUM_EXPDESIGN: + r-base: \$R_VERSION + END_VERSIONS + """ +} \ No newline at end of file diff --git a/modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R b/modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R new file mode 100644 index 0000000..77a7b64 --- /dev/null +++ b/modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R @@ -0,0 +1,69 @@ +# Make a DiMSum experimental design from a deepmutscan samplesheet. +# - samplesheet_csv: path to CSV with columns sample,type,replicate,file1,file2 +# - out_path: where to write the TSV (default "experimentalDesign.tsv") +# Returns: the experimental design as a data.frame +make_dimsum_experimental_design <- function(samplesheet_csv, out_path = "experimentalDesign.tsv") { + # ---- read & normalize ---- + ss <- read.csv(samplesheet_csv, stringsAsFactors = FALSE, check.names = FALSE) + names(ss) <- tolower(names(ss)) + + # tolerate missing file2 column (single-end) + if (!"file2" %in% names(ss)) ss$file2 <- "" + + required <- c("sample", "type", "replicate", "file1", "file2") + missing <- setdiff(required, names(ss)) + if (length(missing) > 0) stop("Samplesheet missing columns: ", paste(missing, collapse = ", ")) + + # coerce types + ss$replicate <- as.integer(ss$replicate) + + # ---- derive sample_name strategy ---- + # If only one biological sample present (e.g. one protein), use "input1", "output2", ... + # If multiple biological samples present, prefix with 'sample' to avoid collisions: + # "GID1A_input1", "GID1B_output2", ... + multi_base <- length(unique(ss$sample)) > 1 + if (multi_base) { + sample_name <- paste(ss$sample, ss$type, ss$replicate, sep = "_") + } else { + sample_name <- paste0(ss$type, ss$replicate) + } + + # ---- build DiMSum columns ---- + experiment_replicate <- ss$replicate + selection_id <- ifelse(ss$type == "input", 0L, + ifelse(ss$type == "output", 1L, NA_integer_)) + # assume one selection batch + selection_replicate <- ifelse(ss$type == "output", 1L, NA_integer_) + # assume one technical batch + technical_replicate <- rep(1L, nrow(ss)) + + pair1 <- basename(ss$file1) + # keep empty string for single-end / missing file2 + pair2 <- ifelse(is.na(ss$file2) | ss$file2 == "", "", basename(ss$file2)) + + ed <- data.frame( + sample_name = sample_name, + experiment_replicate = experiment_replicate, + selection_id = selection_id, + selection_replicate = selection_replicate, + technical_replicate = technical_replicate, + pair1 = pair1, + pair2 = pair2, + stringsAsFactors = FALSE + ) + + # ---- order rows: by sample (if multiple), type (input, output, quality), then replicate ---- + type_rank <- match(ss$type, c("input", "output", "quality")) + ord <- if (multi_base) { + order(ss$sample, type_rank, ss$replicate, na.last = TRUE) + } else { + order(type_rank, ss$replicate, na.last = TRUE) + } + ed <- ed[ord, , drop = FALSE] + rownames(ed) <- NULL + + # ---- write & return ---- + write.table(ed, file = out_path, sep = "\t", row.names = FALSE, col.names = TRUE, quote = FALSE, na = "") + return(ed) +} + diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index 67ce1d4..a77be96 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -21,6 +21,7 @@ include { VISUALIZATION_LOGDIFF } from '../modules/local/visualization/visu include { VISUALIZATION_SEQDEPTH } from '../modules/local/visualization/visualization' include { GATK_GATKTODIMSUM } from '../modules/local/gatk/gatktodimsum' include { MERGE_COUNTS } from '../modules/local/dimsum/merge_counts' +include { EXPDESIGN_DIMSUM } from '../modules/local/dimsum/dimsum_experimental_design' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -83,6 +84,11 @@ Channel .value(params.run_seqdepth) .set { run_seqdepth_ch } +// The deepmutscan samplesheet file provided via --input +Channel + .fromPath(params.input, checkIfExists: true) + .set { ch_samplesheet_csv } + // Define R scripts as channels // Helper to anchor to the repo root @@ -108,6 +114,7 @@ R("modules/local/dmsanalysis/bin/prepare_gatk_data_for_count_heatmaps.R").set { R("modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R").set { prepare_fitness_heatmap_script_ch } R("modules/local/dmsanalysis/bin/process_raw_gatk.R").set { process_raw_gatk_script_ch } R("modules/local/dmsanalysis/bin/merge_counts.R").set { merge_counts_script_ch } +R("modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R").set { exp_design_ch } @@ -404,6 +411,14 @@ if (params.dimsum) { } +if (params.dimsum) { + EXPDESIGN_DIMSUM( + ch_samplesheet_csv, // path to CSV + exp_design_ch // path to R script + ) +} + + emit: From 57c041ffad21e2cf06f5f6c1745a54a0c0791ef6 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Fri, 12 Sep 2025 09:08:24 +0200 Subject: [PATCH 259/317] catch high-count 2nt syn variant --- conf/modules.config | 8 ++ .../local/dimsum/find_synonymous_mutation.nf | 39 +++++++++ modules/local/dimsum/merge_counts.nf | 3 +- .../local/dmsanalysis/bin/find_syn_mutation.R | 87 +++++++++++++++++++ workflows/deepmutscan.nf | 85 +++++++++++------- 5 files changed, 187 insertions(+), 35 deletions(-) create mode 100644 modules/local/dimsum/find_synonymous_mutation.nf create mode 100644 modules/local/dmsanalysis/bin/find_syn_mutation.R diff --git a/conf/modules.config b/conf/modules.config index de026df..bd84c54 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -147,4 +147,12 @@ process { saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } + + withName: 'FIND_SYNONYMOUS_MUTATION' { + publishDir = [ + path: "${params.outdir}/DiMSum", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } } diff --git a/modules/local/dimsum/find_synonymous_mutation.nf b/modules/local/dimsum/find_synonymous_mutation.nf new file mode 100644 index 0000000..a70bc9b --- /dev/null +++ b/modules/local/dimsum/find_synonymous_mutation.nf @@ -0,0 +1,39 @@ +process FIND_SYNONYMOUS_MUTATION { + tag { sample.sample } + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" + + input: + tuple val(sample), path(counts_merged) // from MERGE_COUNTS.out.merged_counts + path wt_fasta // broadcast singleton + val pos_range // "start-end", broadcast singleton + path script // find_syn_mutation.R, broadcast singleton + + output: + tuple val(sample), path("synonymous_wt.txt"), emit: synonymous_wt + path "versions.yml", emit: versions + + script: + """ + set -euo pipefail + start_stop_codon="$pos_range" + + Rscript -e "source('$script'); \ + seq <- pick_synonymous_wt_from_range( \ + wt_fasta='$wt_fasta', \ + counts_merged_tsv='$counts_merged', \ + pos_range='\$start_stop_codon' \ + ); \ + write(seq, file='synonymous_wt.txt')" + + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + cat <<-END_VERSIONS > versions.yml + FIND_SYNONYMOUS_MUTATION: + r-base: \$R_VERSION + END_VERSIONS + """ +} diff --git a/modules/local/dimsum/merge_counts.nf b/modules/local/dimsum/merge_counts.nf index 9d01526..7459a4e 100644 --- a/modules/local/dimsum/merge_counts.nf +++ b/modules/local/dimsum/merge_counts.nf @@ -1,7 +1,6 @@ process MERGE_COUNTS { tag "${sample.sample}" label 'process_single' - publishDir "${params.outdir}/dimsum/${sample.sample}", mode: 'copy' conda "${moduleDir}/environment.yml" container "${ workflow.containerEngine == 'singularity' @@ -13,7 +12,7 @@ process MERGE_COUNTS { path merge_script output: - path "counts_merged.tsv", emit: merged_counts + tuple val(sample), path("counts_merged.tsv"), emit: merged_counts path "versions.yml", emit: versions script: diff --git a/modules/local/dmsanalysis/bin/find_syn_mutation.R b/modules/local/dmsanalysis/bin/find_syn_mutation.R new file mode 100644 index 0000000..716f4f9 --- /dev/null +++ b/modules/local/dmsanalysis/bin/find_syn_mutation.R @@ -0,0 +1,87 @@ +suppressMessages(library(Biostrings)) + +# Pick a synonymous "WT substitute" for DiMSum normalization using a fixed coding window. +# Inputs: +# wt_fasta : path to FASTA (single WT sequence) +# counts_merged_tsv : path to merged counts (columns: nt_seq, input1..N, output1..M) +# pos_range : "start-end" (1-based, inclusive), e.g. "352-1383" +# Returns: +# character scalar: chosen nt sequence +# +# Preference: +# 1) AA-identical (fully synonymous) AND exactly 2 nt mismatches vs WT, both within ONE codon. +# 2) If none, AA-identical AND exactly 1 nt mismatch vs WT. +# 3) If more than one, pick highest mean of input counts. +# 4) If still none, stop with an error. + +pick_synonymous_wt_from_range <- function(wt_fasta, counts_merged_tsv, pos_range) { + ## ---- parse range ---- + pr <- strsplit(as.character(pos_range), "-", fixed = TRUE)[[1]] + if (length(pr) != 2L) stop("pos_range must be 'start-end', got: ", pos_range) + start_pos <- as.integer(pr[1]); end_pos <- as.integer(pr[2]) + if (is.na(start_pos) || is.na(end_pos) || start_pos < 1L || end_pos < start_pos) + stop("Invalid pos_range: ", pos_range) + + ## ---- WT window ---- + wt_set <- Biostrings::readDNAStringSet(wt_fasta) + if (length(wt_set) != 1L) stop("WT FASTA must contain exactly one sequence.") + wt_subseq <- Biostrings::subseq(wt_set[[1]], start = start_pos, end = end_pos) + wt_seq_chr <- as.character(wt_subseq) + wt_len <- nchar(wt_seq_chr) + if ((wt_len %% 3) != 0) stop("Provided window length is not divisible by 3: ", wt_len) + wt_aa <- Biostrings::translate(wt_subseq, if.fuzzy.codon = "X") + wt_chars <- strsplit(wt_seq_chr, "", fixed = TRUE)[[1]] + + ## ---- counts ---- + df <- utils::read.delim(counts_merged_tsv, sep = "\t", header = TRUE, + stringsAsFactors = FALSE, check.names = FALSE) + if (!"nt_seq" %in% names(df)) stop("counts_merged_tsv must have a 'nt_seq' column.") + + df$nt_seq <- toupper(df$nt_seq) + keep_len <- nchar(df$nt_seq) == wt_len + if (!any(keep_len)) stop("No sequences match WT window length (", wt_len, ").") + if (!all(keep_len)) df <- df[keep_len, , drop = FALSE] + + # input columns & mean (works with 1+ replicates) + input_cols <- grep("^input", names(df), value = TRUE) + if (length(input_cols) == 0L) stop("No input columns found (expect names starting with 'input').") + input_mat <- as.data.frame(lapply(df[, input_cols, drop = FALSE], function(x) as.numeric(as.character(x)))) + input_mean <- if (length(input_cols) == 1L) input_mat[[1]] else rowMeans(as.matrix(input_mat), na.rm = TRUE) + + ## ---- synonymous filter ---- + var_set <- Biostrings::DNAStringSet(df$nt_seq) + var_aa <- Biostrings::translate(var_set, if.fuzzy.codon = "X") + syn_idx <- which(as.character(var_aa) == as.character(wt_aa)) + if (length(syn_idx) == 0L) stop("No fully-synonymous variants found relative to WT translation.") + + # helpers + mismatch_positions <- function(seq_nt_chars) which(seq_nt_chars != wt_chars) # 1-based positions + codon_index <- function(pos_vec) floor((pos_vec - 1L) / 3L) # 0-based codon bin + + # preference 1: exactly 2 mismatches, both within the same codon + cand_two_one <- Filter(function(i) { + vchars <- strsplit(df$nt_seq[i], "", fixed = TRUE)[[1]] + pos <- mismatch_positions(vchars) + length(pos) == 2L && length(unique(codon_index(pos))) == 1L + }, syn_idx) + + choose_best <- function(idx_vec) idx_vec[ which.max(input_mean[idx_vec]) ] + + if (length(cand_two_one) > 0L) { + best_i <- choose_best(cand_two_one) + return(as.character(df$nt_seq[best_i])) + } + + # preference 2 (fallback): exactly 1 mismatch (still synonymous) + cand_one <- Filter(function(i) { + vchars <- strsplit(df$nt_seq[i], "", fixed = TRUE)[[1]] + length(mismatch_positions(vchars)) == 1L + }, syn_idx) + + if (length(cand_one) > 0L) { + best_i <- choose_best(cand_one) + return(as.character(df$nt_seq[best_i])) + } + + stop("No suitable synonymous variant found: neither 2-in-1-codon nor 1-nt synonymous candidates present.") +} diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index a77be96..cc6d62b 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -22,6 +22,7 @@ include { VISUALIZATION_SEQDEPTH } from '../modules/local/visualization/vis include { GATK_GATKTODIMSUM } from '../modules/local/gatk/gatktodimsum' include { MERGE_COUNTS } from '../modules/local/dimsum/merge_counts' include { EXPDESIGN_DIMSUM } from '../modules/local/dimsum/dimsum_experimental_design' +include { FIND_SYNONYMOUS_MUTATION } from '../modules/local/dimsum/find_synonymous_mutation' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -115,6 +116,7 @@ R("modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R").set { R("modules/local/dmsanalysis/bin/process_raw_gatk.R").set { process_raw_gatk_script_ch } R("modules/local/dmsanalysis/bin/merge_counts.R").set { merge_counts_script_ch } R("modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R").set { exp_design_ch } +R("modules/local/dmsanalysis/bin/find_syn_mutation.R").set { syn_mut_ch } @@ -377,50 +379,67 @@ logdiff_scriptN = fanoutTo(library_completed_variantCounts_ch, lo -// ----- DiMSum: group per biological sample (from samplesheet) and merge counts to use for DiMSum input ----- + // ----- DiMSum: group per biological sample (from samplesheet) and merge counts to use for DiMSum input ----- -GATK_GATKTODIMSUM.out.dimsum_input - .map { meta, tsv -> - def s = meta.sample as String - def id = meta.id as String - def base = s ? (s.replaceFirst(/_(input|output|quality)\d+$/, '')) + GATK_GATKTODIMSUM.out.dimsum_input + .map { meta, tsv -> + def s = meta.sample as String + def id = meta.id as String + def base = s ? (s.replaceFirst(/_(input|output|quality)\d+$/, '')) : (id?.tokenize('_')?.first()) - tuple(base as String, tuple(meta, tsv)) - } - .groupTuple() - .map { base, pairs -> - def metas = pairs.collect { it[0] } - def inputs = pairs.findAll { it[0].type == 'input' }.sort { it[0].replicate }.collect { it[1] } - def outputs = pairs.findAll { it[0].type == 'output' }.sort { it[0].replicate }.collect { it[1] } - tuple([sample: base], metas, inputs, outputs) - } - .filter { smeta, metas, ins, outs -> ins && outs } - .set { ch_dimsum_bundled } - -// Broadcast the singleton script path to match each bundle -def ch_merge_script_for_each = merge_counts_script_ch - .combine(ch_dimsum_bundled) - .map { it[0] } // keep the script path, one per bundle - -// Launch the merge -if (params.dimsum) { - MERGE_COUNTS( - ch_dimsum_bundled, // tuple val(sample), val(metas), path(input_counts), path(output_counts) - ch_merge_script_for_each // path merge_script (broadcast) - ) -} + tuple(base as String, tuple(meta, tsv)) + } + .groupTuple() + .map { base, pairs -> + def metas = pairs.collect { it[0] } + def inputs = pairs.findAll { it[0].type == 'input' }.sort { it[0].replicate }.collect { it[1] } + def outputs = pairs.findAll { it[0].type == 'output' }.sort { it[0].replicate }.collect { it[1] } + tuple([sample: base], metas, inputs, outputs) + } + .filter { smeta, metas, ins, outs -> ins && outs } + .set { ch_dimsum_bundled } + + // Broadcast the singleton script path to match each bundle + def ch_merge_script_for_each = merge_counts_script_ch + .combine(ch_dimsum_bundled) + .map { it[0] } // keep the script path, one per bundle + + // Launch the merge of counts in DiMSum input format + if (params.dimsum) { + MERGE_COUNTS( + ch_dimsum_bundled, // tuple val(sample), val(metas), path(input_counts), path(output_counts) + ch_merge_script_for_each // path merge_script (broadcast) + ) + } + + // Create experimental design file to use for DiMSum + if (params.dimsum) { + EXPDESIGN_DIMSUM( + ch_samplesheet_csv, // path to CSV + exp_design_ch // path to R script + ) + } + +// --- Synonymous WT selection (runs only when --dimsum) --- +// Strip meta once: keep only the fasta path +ch_fasta.map { it[1] }.set { ch_fasta_path } // path(/…/GID1A.fasta) if (params.dimsum) { - EXPDESIGN_DIMSUM( - ch_samplesheet_csv, // path to CSV - exp_design_ch // path to R script + // MERGE_COUNTS.out.merged_counts shape: tuple( val([sample:'GID1A']), path("counts_merged.tsv") ) + FIND_SYNONYMOUS_MUTATION( + MERGE_COUNTS.out.merged_counts, // tuple(val(sample), path counts_merged.tsv) + ch_fasta_path.combine(MERGE_COUNTS.out.merged_counts).map { it[0] }, // path wt_fasta (broadcast to N) + reading_frame_ch.combine(MERGE_COUNTS.out.merged_counts).map { it[0] }, // val pos_range (broadcast) + syn_mut_ch.combine(MERGE_COUNTS.out.merged_counts).map { it[0] } // path R script (broadcast) ) } + + emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From 65a861e4358f03fe9eeadced187c014bc59387fd Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Fri, 12 Sep 2025 10:21:30 +0200 Subject: [PATCH 260/317] dimsum (No Docker container -> only use on cluster) --- conf/modules.config | 8 ++++++ modules/local/dimsum/run_dimsum.nf | 46 ++++++++++++++++++++++++++++++ workflows/deepmutscan.nf | 39 +++++++++++++++++++++++++ 3 files changed, 93 insertions(+) create mode 100644 modules/local/dimsum/run_dimsum.nf diff --git a/conf/modules.config b/conf/modules.config index bd84c54..fea4bc0 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -155,4 +155,12 @@ process { saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } + + withName: 'RUN_DIMSUM' { + publishDir = [ + path: "${params.outdir}/DiMSum", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } } diff --git a/modules/local/dimsum/run_dimsum.nf b/modules/local/dimsum/run_dimsum.nf new file mode 100644 index 0000000..48ddc05 --- /dev/null +++ b/modules/local/dimsum/run_dimsum.nf @@ -0,0 +1,46 @@ +process RUN_DIMSUM { + tag { sample.sample } + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' + ? 'oras://community.wave.seqera.io/library/r-dimsum:1.4--4357734d345c8ccc' + : 'oras://community.wave.seqera.io/library/r-dimsum:1.4--4357734d345c8ccc' }" + + input: + tuple val(sample), path(counts_merged) + path(wt_txt) + path(exp_design) + + output: + path "reports/**", emit: reports_folder + path "dimsum_results**", emit: results_dir + path "report.html", emit: report + path "tmp/**", emit: tmp_folder + path "versions.yml", emit: versions + + script: + """ + set -euo pipefail + + # DiMSum expects the sequence string, not a file path + WT=\$(tr -d ' \r\\n\\t' < "!{wt_txt}") + + ( DiMSum \\ + --experimentDesignPath "$exp_design" \\ + --wildtypeSequence "\$WT" \\ + --countPath "$counts_merged" \\ + --startStage 4 \\ + --stopStage 5 \\ + --retainIntermediateFiles T \\ + --projectName "dimsum_results" \\ + --fastqFileDir . \\ + ) + + R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') + cat <<-END_VERSIONS > versions.yml + DIMSUM_RUN: + r-base: \$R_VERSION + END_VERSIONS + """ +} diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index cc6d62b..d9ef1a5 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -23,6 +23,7 @@ include { GATK_GATKTODIMSUM } from '../modules/local/gatk/gatktodimsum' include { MERGE_COUNTS } from '../modules/local/dimsum/merge_counts' include { EXPDESIGN_DIMSUM } from '../modules/local/dimsum/dimsum_experimental_design' include { FIND_SYNONYMOUS_MUTATION } from '../modules/local/dimsum/find_synonymous_mutation' +include { RUN_DIMSUM } from '../modules/local/dimsum/run_dimsum' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -436,6 +437,44 @@ if (params.dimsum) { } +// --- DiMSum execution (only when --dimsum) --- +if (params.dimsum) { + // Shapes: + // MERGE_COUNTS.out.merged_counts -> tuple(val([sample:'GID1A']), path('counts_merged.tsv')) + // FIND_SYNONYMOUS_MUTATION.out.synonymous_wt -> tuple(val([sample:'GID1A']), path('synonymous_wt.txt')) + // EXPDESIGN_DIMSUM.out.experimental_design -> path('experimentalDesign.tsv') (singleton) + + // 1) Key counts and WT by biological sample name to align them robustly + def ch_counts_keyed_d = MERGE_COUNTS.out.merged_counts + .map { smp, counts -> tuple(smp.sample as String, smp, counts) } + + def ch_wt_keyed_d = FIND_SYNONYMOUS_MUTATION.out.synonymous_wt + .map { smp, wt -> tuple(smp.sample as String, wt) } + + // 2) Join by key -> (val(sample), path(counts), path(wt)) + def ch_counts_wt_d = ch_counts_keyed_d.join(ch_wt_keyed_d) + .map { key, smp, counts, wt -> tuple(smp, counts, wt) } + + // 3) Broadcast experimental design (singleton) to each sample triple + def ch_exp_for_each_d = EXPDESIGN_DIMSUM.out.experimental_design + .combine(ch_counts_wt_d) + .map { it[0] } + + // 4) Final aligned channels for RUN_DIMSUM + def ch_run_counts_d = ch_counts_wt_d.map { smp, counts, wt -> tuple(smp, counts) } // matches: tuple val(sample), path(counts_merged) + def ch_run_wt_d = ch_counts_wt_d.map { smp, counts, wt -> wt } // matches: path(wt_txt) + def ch_run_exp_d = ch_exp_for_each_d // matches: path(exp_design) + + // 5) Launch + RUN_DIMSUM( + ch_run_counts_d, // tuple val(sample), path(counts_merged) + ch_run_wt_d, // path wt_txt + ch_run_exp_d // path experimentalDesign.tsv + ) +} + + + From a344d903f6d286a4547bc05fbce030fce7ccbcf3 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Sun, 14 Sep 2025 23:01:29 +0200 Subject: [PATCH 261/317] Update nextflow.config --- nextflow.config | 16 ++++++++-------- 1 file changed, 8 insertions(+), 8 deletions(-) diff --git a/nextflow.config b/nextflow.config index ebf888f..67771ec 100644 --- a/nextflow.config +++ b/nextflow.config @@ -246,14 +246,14 @@ env { } // Set bash options -process.shell = """\ -bash - -set -e # Exit if a tool returns a non-zero status/exit code -set -u # Treat unset variables and parameters as an error -set -o pipefail # Returns the status of the last command to exit with a non-zero status or zero if all successfully execute -set -C # No clobber - prevent output redirection from overwriting files. -""" +process.shell = [ + "bash", + "-C", // No clobber - prevent output redirection from overwriting files. + "-e", // Exit if a tool returns a non-zero status/exit code + "-u", // Treat unset variables and parameters as an error + "-o", // Returns the status of the last command to exit.. + "pipefail" // ..with a non-zero status or zero if all successfully execute +] // Disable process selector warnings by default. Use debug profile to enable warnings. nextflow.enable.configProcessNamesValidation = false From 58b22f095738b5d63e0f7b22ea363a6a8dbb2d53 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Mon, 15 Sep 2025 00:17:35 +0200 Subject: [PATCH 262/317] call of wt_seq --- modules/local/dimsum/run_dimsum.nf | 23 +++++++++++------------ 1 file changed, 11 insertions(+), 12 deletions(-) diff --git a/modules/local/dimsum/run_dimsum.nf b/modules/local/dimsum/run_dimsum.nf index 48ddc05..8b58d0a 100644 --- a/modules/local/dimsum/run_dimsum.nf +++ b/modules/local/dimsum/run_dimsum.nf @@ -5,7 +5,7 @@ process RUN_DIMSUM { conda "${moduleDir}/environment.yml" container "${ workflow.containerEngine == 'singularity' ? 'oras://community.wave.seqera.io/library/r-dimsum:1.4--4357734d345c8ccc' - : 'oras://community.wave.seqera.io/library/r-dimsum:1.4--4357734d345c8ccc' }" + : 'docker.io/bwehnert1008/dms_qc_dimsum_environment@sha256:08f3bd8441df7b4a7e05aadeca178862153cf723e64097a48a2744b2698b15dd' }" input: tuple val(sample), path(counts_merged) @@ -24,18 +24,17 @@ process RUN_DIMSUM { set -euo pipefail # DiMSum expects the sequence string, not a file path - WT=\$(tr -d ' \r\\n\\t' < "!{wt_txt}") + WT=\$(tr -d ' \r\\n\\t' < "$wt_txt") - ( DiMSum \\ - --experimentDesignPath "$exp_design" \\ - --wildtypeSequence "\$WT" \\ - --countPath "$counts_merged" \\ - --startStage 4 \\ - --stopStage 5 \\ - --retainIntermediateFiles T \\ - --projectName "dimsum_results" \\ - --fastqFileDir . \\ - ) + DiMSum \ + --experimentDesignPath "$exp_design" \ + --wildtypeSequence "\$WT" \ + --countPath "$counts_merged" \ + --startStage 4 \ + --stopStage 5 \ + --retainIntermediateFiles T \ + --projectName "dimsum_results" \ + --fastqFileDir . \ R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') cat <<-END_VERSIONS > versions.yml From 99ad79247fb4eb8f8c1aa3dad79c8a45a2419de0 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Mon, 15 Sep 2025 00:20:17 +0200 Subject: [PATCH 263/317] Update nextflow.config --- nextflow.config | 16 ++++++++-------- 1 file changed, 8 insertions(+), 8 deletions(-) diff --git a/nextflow.config b/nextflow.config index ebf888f..67771ec 100644 --- a/nextflow.config +++ b/nextflow.config @@ -246,14 +246,14 @@ env { } // Set bash options -process.shell = """\ -bash - -set -e # Exit if a tool returns a non-zero status/exit code -set -u # Treat unset variables and parameters as an error -set -o pipefail # Returns the status of the last command to exit with a non-zero status or zero if all successfully execute -set -C # No clobber - prevent output redirection from overwriting files. -""" +process.shell = [ + "bash", + "-C", // No clobber - prevent output redirection from overwriting files. + "-e", // Exit if a tool returns a non-zero status/exit code + "-u", // Treat unset variables and parameters as an error + "-o", // Returns the status of the last command to exit.. + "pipefail" // ..with a non-zero status or zero if all successfully execute +] // Disable process selector warnings by default. Use debug profile to enable warnings. nextflow.enable.configProcessNamesValidation = false From 4a4e0904b370d4aca7a2a0d13c2323acf410d6b5 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Mon, 15 Sep 2025 00:30:48 +0200 Subject: [PATCH 264/317] Update modules.config --- conf/modules.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/modules.config b/conf/modules.config index fea4bc0..1d667bd 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -158,7 +158,7 @@ process { withName: 'RUN_DIMSUM' { publishDir = [ - path: "${params.outdir}/DiMSum", + path: "${params.outdir}/DiMSum/output", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] From f3e3146fc3e77aed32a9080b6dc19424254cc053 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 15 Sep 2025 09:04:21 +0200 Subject: [PATCH 265/317] Update dimsum_experimentalDesign.R --- modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R b/modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R index 77a7b64..123593d 100644 --- a/modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R +++ b/modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R @@ -23,7 +23,7 @@ make_dimsum_experimental_design <- function(samplesheet_csv, out_path = "experim # "GID1A_input1", "GID1B_output2", ... multi_base <- length(unique(ss$sample)) > 1 if (multi_base) { - sample_name <- paste(ss$sample, ss$type, ss$replicate, sep = "_") + sample_name <- paste(ss$sample, ss$type, ss$replicate, sep = "") } else { sample_name <- paste0(ss$type, ss$replicate) } From f6850004e560721f49bf151350e0cb5749ba383a Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 15 Sep 2025 19:06:31 +0200 Subject: [PATCH 266/317] Update run_dimsum.nf --- modules/local/dimsum/run_dimsum.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/dimsum/run_dimsum.nf b/modules/local/dimsum/run_dimsum.nf index 8b58d0a..508db30 100644 --- a/modules/local/dimsum/run_dimsum.nf +++ b/modules/local/dimsum/run_dimsum.nf @@ -34,7 +34,7 @@ process RUN_DIMSUM { --stopStage 5 \ --retainIntermediateFiles T \ --projectName "dimsum_results" \ - --fastqFileDir . \ + --fastqFileDir . R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') cat <<-END_VERSIONS > versions.yml From 4fbb8f49980e9ef9a31fd72709a0b582acbaca8c Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 15 Sep 2025 20:53:10 +0200 Subject: [PATCH 267/317] Update run_dimsum.nf --- modules/local/dimsum/run_dimsum.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/modules/local/dimsum/run_dimsum.nf b/modules/local/dimsum/run_dimsum.nf index 508db30..d2447e4 100644 --- a/modules/local/dimsum/run_dimsum.nf +++ b/modules/local/dimsum/run_dimsum.nf @@ -32,6 +32,7 @@ process RUN_DIMSUM { --countPath "$counts_merged" \ --startStage 4 \ --stopStage 5 \ + --fitnessErrorModel F \ --retainIntermediateFiles T \ --projectName "dimsum_results" \ --fastqFileDir . From 9285b156da0429bf4b909c78ffe210f8b0299445 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 15 Sep 2025 20:53:19 +0200 Subject: [PATCH 268/317] Update run_dimsum.nf --- modules/local/dimsum/run_dimsum.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/dimsum/run_dimsum.nf b/modules/local/dimsum/run_dimsum.nf index d2447e4..aa4e8b7 100644 --- a/modules/local/dimsum/run_dimsum.nf +++ b/modules/local/dimsum/run_dimsum.nf @@ -35,7 +35,7 @@ process RUN_DIMSUM { --fitnessErrorModel F \ --retainIntermediateFiles T \ --projectName "dimsum_results" \ - --fastqFileDir . + --fastqFileDir . \ R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') cat <<-END_VERSIONS > versions.yml From 6407aa0d4115e124b07f8cd37de41a6d80ead478 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Sun, 21 Sep 2025 19:09:01 +0200 Subject: [PATCH 269/317] Update run_dimsum.nf --- modules/local/dimsum/run_dimsum.nf | 3 --- 1 file changed, 3 deletions(-) diff --git a/modules/local/dimsum/run_dimsum.nf b/modules/local/dimsum/run_dimsum.nf index aa4e8b7..9af63b2 100644 --- a/modules/local/dimsum/run_dimsum.nf +++ b/modules/local/dimsum/run_dimsum.nf @@ -13,10 +13,7 @@ process RUN_DIMSUM { path(exp_design) output: - path "reports/**", emit: reports_folder path "dimsum_results**", emit: results_dir - path "report.html", emit: report - path "tmp/**", emit: tmp_folder path "versions.yml", emit: versions script: From 955be99235355f8b279db0ef2cd9c2d062b82bf7 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 27 Oct 2025 17:32:57 +0100 Subject: [PATCH 270/317] Update pipeline_steps.md --- docs/pipeline_steps.md | 13 ++++++------- 1 file changed, 6 insertions(+), 7 deletions(-) diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md index 6b67e99..a93213d 100644 --- a/docs/pipeline_steps.md +++ b/docs/pipeline_steps.md @@ -67,24 +67,23 @@ Custom visualisations allow for inspection of (1) mutation efficiency along the ## 6. Data Summarisation for Fitness Estimation -Steps 1-5 are iteratively run across all samples defined in the `.csv` spreadsheet. Once read alignment, merging, mutation counting, and library QC have been completed for the full list of samples, users can opt to proceed with fitness estimation using a number of tools (default: [**DiMSum**](https://github.com/lehner-lab/DiMSum)). The pipeline generates the tool-specific input files by merging mutation counts across samples, using custom R scripts. +Steps 1-5 are iteratively run across all samples defined in the `.csv` spreadsheet. Once read alignment, merging, mutation counting, and library QC have been completed for the full list of samples, users can opt to proceed with fitness estimation. To this end, the pipeline generates all the necessary input files by merging mutation counts across samples. ------------------------------------------------------------------------ ## 7. Single Nucleotide Variant Error Correction *(in development)* -This module will implement strategies to distinguish true single nucleotide variants from sequencing artefacts. Methods under consideration include: -- Empirical error rate modelling based on false double mutants +This module will implement strategies to distinguish true single nucleotide variants from sequencing artefacts. There are two options to perform this: - Empirical error rate modelling based on wildtype sequencing +- Empirical error rate modelling based on false double mutants *(in development)* ------------------------------------------------------------------------ ## 8. Fitness Estimation *(in development)* -Using DiMSum, the final step of the pipeline will perform fitness estimation based on mutation counts. Planned features include: -- Integration of multiple other fitness inference tools, including [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) -- Options for replicates and condition-specific comparisons -- Standardised output formats for downstream analysis and comparison +The final step of the pipeline will perform fitness estimation based on mutation counts. By default, we calculate fitness scores as the logarithm of variants' output to input ratio, normalised to that of the provided wildtype sequence. Future expansions may include: +- Integration of other popular fitness inference tools, including [DiMSum](https://github.com/lehner-lab/DiMSum), [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) +- Standardised output formats for downstream analyses and comparison > [!IMPORTANT] > We note that exact wildtype sequence reads are filtered out in stage 2. Including synonymous wildtype codons in the original mutagenesis design is therefore essential when it comes to calibrating the fitness calculations. From 9a434c682df5f40529c440f46088394dd1bb6eab Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 27 Oct 2025 17:33:47 +0100 Subject: [PATCH 271/317] Update pipeline_steps.md --- docs/pipeline_steps.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md index a93213d..83c18b5 100644 --- a/docs/pipeline_steps.md +++ b/docs/pipeline_steps.md @@ -82,7 +82,7 @@ This module will implement strategies to distinguish true single nucleotide vari ## 8. Fitness Estimation *(in development)* The final step of the pipeline will perform fitness estimation based on mutation counts. By default, we calculate fitness scores as the logarithm of variants' output to input ratio, normalised to that of the provided wildtype sequence. Future expansions may include: -- Integration of other popular fitness inference tools, including [DiMSum](https://github.com/lehner-lab/DiMSum), [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) +- Integration of other popular fitness inference tools, including [DiMSum](https://github.com/lehner-lab/DiMSum), [Enrich2](https://github.com/FowlerLab/Enrich2), [rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) - Standardised output formats for downstream analyses and comparison > [!IMPORTANT] From 2504375d95505a97e1708be7a45aeab356278e07 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 27 Oct 2025 17:34:07 +0100 Subject: [PATCH 272/317] Update README.md --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 5dece0a..acc6c8f 100644 --- a/README.md +++ b/README.md @@ -28,7 +28,7 @@ While DNA synthesis and sequencing technologies have advanced substantially, lon # 2. Features of nf-core/deepmutscan - End-to-end analyses of DMS shotgun sequencing data - Modular, three-stage workflow: alignment → QC → error-aware fitness estimation -- Integrates with popular statistical tools like [DiMSum](https://github.com/lehner-lab/DiMSum), [Enrich2](https://github.com/FowlerLab/Enrich2), [Rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) +- Integrates with popular statistical tools like [DiMSum](https://github.com/lehner-lab/DiMSum), [Enrich2](https://github.com/FowlerLab/Enrich2), [rosace](https://github.com/pimentellab/rosace/) and [mutscan](https://github.com/fmicompbio/mutscan) - Supports multiple mutagenesis strategies, e.g. nicking by NNK and NNS codons - Containerized via Docker, Singularity and Apptainer - Scalable across HPC and Cloud systems From 5b20a24f6efc2fe5488b8c49d584c54d16306ea2 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Thu, 30 Oct 2025 14:45:55 +0100 Subject: [PATCH 273/317] update folder structure from dimsum to fitness for modules, scripts, and results --- conf/modules.config | 18 ++---- .../{gatk_to_dimsum.R => gatk_to_fitness.R} | 4 +- .../find_synonymous_mutation.nf | 0 .../fitness_experimental_design.nf} | 2 +- .../local/{dimsum => fitness}/merge_counts.nf | 0 .../{gatktodimsum.nf => gatktofitness.nf} | 12 ++-- nextflow_schema.json | 7 ++- workflows/deepmutscan.nf | 56 ++++++++++--------- 8 files changed, 50 insertions(+), 49 deletions(-) rename modules/local/dmsanalysis/bin/{gatk_to_dimsum.R => gatk_to_fitness.R} (98%) rename modules/local/{dimsum => fitness}/find_synonymous_mutation.nf (100%) rename modules/local/{dimsum/dimsum_experimental_design.nf => fitness/fitness_experimental_design.nf} (97%) rename modules/local/{dimsum => fitness}/merge_counts.nf (100%) rename modules/local/gatk/{gatktodimsum.nf => gatktofitness.nf} (71%) diff --git a/conf/modules.config b/conf/modules.config index 1d667bd..db6ed04 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -116,7 +116,7 @@ process { ] } - withName: 'GATK_GATKTODIMSUM' { + withName: 'GATK_GATKTOFITNESS' { publishDir = [ path: "${params.outdir}/DiMSum/single_counts", mode: 'copy', @@ -126,23 +126,15 @@ process { withName: 'MERGE_COUNTS' { publishDir = [ - path: "${params.outdir}/DiMSum", + path: "${params.outdir}/fitness", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] } - withName: 'MERGE_COUNTS' { - publishDir = [ - path: "${params.outdir}/DiMSum", - mode: 'copy', - saveAs: { filename -> filename == 'versions.yml' ? null : filename } - ] - } - - withName: 'EXPDESIGN_DIMSUM' { + withName: 'EXPDESIGN_FITNESS' { publishDir = [ - path: "${params.outdir}/DiMSum", + path: "${params.outdir}/fitness", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] @@ -150,7 +142,7 @@ process { withName: 'FIND_SYNONYMOUS_MUTATION' { publishDir = [ - path: "${params.outdir}/DiMSum", + path: "${params.outdir}/fitness", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] diff --git a/modules/local/dmsanalysis/bin/gatk_to_dimsum.R b/modules/local/dmsanalysis/bin/gatk_to_fitness.R similarity index 98% rename from modules/local/dmsanalysis/bin/gatk_to_dimsum.R rename to modules/local/dmsanalysis/bin/gatk_to_fitness.R index 916404d..5004d97 100644 --- a/modules/local/dmsanalysis/bin/gatk_to_dimsum.R +++ b/modules/local/dmsanalysis/bin/gatk_to_fitness.R @@ -97,7 +97,7 @@ suppressMessages(library(Biostrings)) -generate_dimsum_input <- function(wt_seq_path, gatk_file, pos_range, output_file_path) { +generate_fitness_input <- function(wt_seq_path, gatk_file, pos_range, output_file_path) { # Parse the position range positions <- unlist(strsplit(pos_range, "-")) start_pos <- as.numeric(positions[1]) @@ -162,7 +162,7 @@ generate_dimsum_input <- function(wt_seq_path, gatk_file, pos_range, output_file write.table(processed_data, file = output_file_path, sep = "\t", row.names = FALSE, col.names = FALSE, quote = FALSE) } -# generate_dimsum_input( +# generate_fitness_input( # "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/MORtn5_reference.fa", # "/Users/benjaminwehnert/CRG/DMS_QC/testing_data/gatk_filtered_by_codon_library.csv", # "23-1225", diff --git a/modules/local/dimsum/find_synonymous_mutation.nf b/modules/local/fitness/find_synonymous_mutation.nf similarity index 100% rename from modules/local/dimsum/find_synonymous_mutation.nf rename to modules/local/fitness/find_synonymous_mutation.nf diff --git a/modules/local/dimsum/dimsum_experimental_design.nf b/modules/local/fitness/fitness_experimental_design.nf similarity index 97% rename from modules/local/dimsum/dimsum_experimental_design.nf rename to modules/local/fitness/fitness_experimental_design.nf index 781a768..e6ec6a2 100644 --- a/modules/local/dimsum/dimsum_experimental_design.nf +++ b/modules/local/fitness/fitness_experimental_design.nf @@ -1,4 +1,4 @@ -process EXPDESIGN_DIMSUM { +process EXPDESIGN_FITNESS { tag "experimentalDesign" label 'process_single' diff --git a/modules/local/dimsum/merge_counts.nf b/modules/local/fitness/merge_counts.nf similarity index 100% rename from modules/local/dimsum/merge_counts.nf rename to modules/local/fitness/merge_counts.nf diff --git a/modules/local/gatk/gatktodimsum.nf b/modules/local/gatk/gatktofitness.nf similarity index 71% rename from modules/local/gatk/gatktodimsum.nf rename to modules/local/gatk/gatktofitness.nf index f23335f..3351c1b 100644 --- a/modules/local/gatk/gatktodimsum.nf +++ b/modules/local/gatk/gatktofitness.nf @@ -1,4 +1,4 @@ -process GATK_GATKTODIMSUM { +process GATK_GATKTOFITNESS { tag "$meta.id" label 'process_single' @@ -14,7 +14,7 @@ process GATK_GATKTODIMSUM { path script // R script output: - tuple val(meta), path("${meta.id}_dimsum_input.tsv"), emit: dimsum_input + tuple val(meta), path("${meta.id}_fitness_input.tsv"), emit: fitness_input path "versions.yml", emit: versions when: @@ -24,19 +24,19 @@ process GATK_GATKTODIMSUM { """ start_stop_codon="$pos_range" - Rscript -e "source('$script'); generate_dimsum_input('$wt_seq', '$variantCounts_filtered_by_library', '\$start_stop_codon', '${meta.id}_dimsum_input.tsv')" + Rscript -e "source('$script'); generate_fitness_input('$wt_seq', '$variantCounts_filtered_by_library', '\$start_stop_codon', '${meta.id}_fitness_input.tsv')" R_VERSION=\$(R --version | head -n 1 | sed -E 's/^R version ([0-9.]+).*/\\1/') cat <<-END_VERSIONS > versions.yml - GATK_GATKTODIMSUM: + GATK_GATKTOFITNESS: r-base: \$R_VERSION END_VERSIONS """ stub: """ - touch ${meta.id}_dimsum_input.tsv - echo "GATK_GATKTODIMSUM:" > versions.yml + touch ${meta.id}_fitness_input.tsv + echo "GATK_GATKTOFITNESS:" > versions.yml echo " stub-version: 0.0.0" >> versions.yml """ } diff --git a/nextflow_schema.json b/nextflow_schema.json index f1fd31e..9a52fb4 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -66,7 +66,12 @@ "dimsum": { "type": "boolean", "default": false, - "description": "Enable DiMSum preparation (group and merge counts per sample into counts_merged.tsv)." + "description": "Enable DiMSum execution." + }, + "fitness": { + "type": "boolean", + "default": false, + "description": "Enable basic fitness calculation and preceded data preparation (group and merge counts per sample into counts_merged.tsv, create a specific experimental design file and find a high-count 2nt synonymous mutation). All data prepared serves also as input for the dimsum fitness calculator." }, "run_seqdepth": { "type": "boolean", diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index d9ef1a5..3ea467d 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -19,10 +19,10 @@ include { VISUALIZATION_GLOBAL_POS_BIASES_COUNTS } from '../modules/local/v include { VISUALIZATION_GLOBAL_POS_BIASES_COV } from '../modules/local/visualization/visualization' include { VISUALIZATION_LOGDIFF } from '../modules/local/visualization/visualization' include { VISUALIZATION_SEQDEPTH } from '../modules/local/visualization/visualization' -include { GATK_GATKTODIMSUM } from '../modules/local/gatk/gatktodimsum' -include { MERGE_COUNTS } from '../modules/local/dimsum/merge_counts' -include { EXPDESIGN_DIMSUM } from '../modules/local/dimsum/dimsum_experimental_design' -include { FIND_SYNONYMOUS_MUTATION } from '../modules/local/dimsum/find_synonymous_mutation' +include { GATK_GATKTOFITNESS } from '../modules/local/gatk/gatktofitness' +include { MERGE_COUNTS } from '../modules/local/fitness/merge_counts' +include { EXPDESIGN_FITNESS } from '../modules/local/fitness/fitness_experimental_design' +include { FIND_SYNONYMOUS_MUTATION } from '../modules/local/fitness/find_synonymous_mutation' include { RUN_DIMSUM } from '../modules/local/dimsum/run_dimsum' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -42,6 +42,7 @@ params.custom_codon_library = params.custom_codon_library ?: '/NULL' params.sliding_window_size = params.sliding_window_size ?: 10 // sliding window size to flatten graphs in plots (e.g. GLOBAL_POS_BIASES_COUNTS function) params.aimed_cov = params.aimed_cov ?: 100 // aimed coverage (assuming equal spread) to visualize threshold in plots params.run_seqdepth = params.run_seqdepth ?: false // creating seqdepth simulation plot, is computationally quite heavy. per default disabled. +params.fitness = params.fitness ?: false // run basic fitness calculation from selection input & output samples params.dimsum = params.dimsum ?: false // run DiMSum for fitness/functionality scores from selection input & output samples @@ -105,7 +106,7 @@ R("modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R").set { counts_per_cov R("modules/local/dmsanalysis/bin/detect_codons.R").set { detect_codons_script_ch } R("modules/local/dmsanalysis/bin/filter_gatk_by_codon_library.R").set { filter_by_library_script_ch } R("modules/local/dmsanalysis/bin/fitness_heatmap.R").set { fitness_heatmap_script_ch } -R("modules/local/dmsanalysis/bin/gatk_to_dimsum.R").set { gatk_to_dimsum_script_ch } +R("modules/local/dmsanalysis/bin/gatk_to_fitness.R").set { gatk_to_fitness_script_ch } R("modules/local/dmsanalysis/bin/global_position_biases_counts_and_counts_per_cov.R").set { global_bias_counts_cov_script_ch } R("modules/local/dmsanalysis/bin/global_position_biases_cov.R").set { global_bias_cov_script_ch } R("modules/local/dmsanalysis/bin/install_packages.R").set { install_packages_script_ch } @@ -366,23 +367,23 @@ logdiff_scriptN = fanoutTo(library_completed_variantCounts_ch, lo } // Broadcast singletons to N (one per sample), anchored on variantCounts_filtered_by_library_ch - ch_fasta_for_dimsum = ch_fasta.combine(variantCounts_filtered_by_library_ch).map { it[1] } // path(fasta) -- N - ch_rf_for_dimsum = reading_frame_ch.combine(variantCounts_filtered_by_library_ch).map { it[0] } // val(range) -- N - ch_script_for_dimsum = gatk_to_dimsum_script_ch.combine(variantCounts_filtered_by_library_ch).map { it[0] } // path(script) -- N + ch_fasta_for_fitness = ch_fasta.combine(variantCounts_filtered_by_library_ch).map { it[1] } // path(fasta) -- N + ch_rf_for_fitness = reading_frame_ch.combine(variantCounts_filtered_by_library_ch).map { it[0] } // val(range) -- N + ch_script_for_fitness = gatk_to_fitness_script_ch.combine(variantCounts_filtered_by_library_ch).map { it[0] } // path(script) -- N // Call with aligned inputs - GATK_GATKTODIMSUM( + GATK_GATKTOFITNESS( variantCounts_filtered_by_library_ch, // tuple(val(meta), path) - ch_fasta_for_dimsum, // path(fasta) - ch_rf_for_dimsum, // val(reading_frame) - ch_script_for_dimsum // path(R script) + ch_fasta_for_fitness, // path(fasta) + ch_rf_for_fitness, // val(reading_frame) + ch_script_for_fitness // path(R script) ) // ----- DiMSum: group per biological sample (from samplesheet) and merge counts to use for DiMSum input ----- - GATK_GATKTODIMSUM.out.dimsum_input + GATK_GATKTOFITNESS.out.fitness_input .map { meta, tsv -> def s = meta.sample as String def id = meta.id as String @@ -398,24 +399,24 @@ logdiff_scriptN = fanoutTo(library_completed_variantCounts_ch, lo tuple([sample: base], metas, inputs, outputs) } .filter { smeta, metas, ins, outs -> ins && outs } - .set { ch_dimsum_bundled } + .set { ch_fitness_bundled } // Broadcast the singleton script path to match each bundle def ch_merge_script_for_each = merge_counts_script_ch - .combine(ch_dimsum_bundled) + .combine(ch_fitness_bundled) .map { it[0] } // keep the script path, one per bundle // Launch the merge of counts in DiMSum input format - if (params.dimsum) { + if (params.fitness) { MERGE_COUNTS( - ch_dimsum_bundled, // tuple val(sample), val(metas), path(input_counts), path(output_counts) + ch_fitness_bundled, // tuple val(sample), val(metas), path(input_counts), path(output_counts) ch_merge_script_for_each // path merge_script (broadcast) ) } // Create experimental design file to use for DiMSum - if (params.dimsum) { - EXPDESIGN_DIMSUM( + if (params.fitness) { + EXPDESIGN_FITNESS( ch_samplesheet_csv, // path to CSV exp_design_ch // path to R script ) @@ -426,7 +427,7 @@ logdiff_scriptN = fanoutTo(library_completed_variantCounts_ch, lo // Strip meta once: keep only the fasta path ch_fasta.map { it[1] }.set { ch_fasta_path } // path(/…/GID1A.fasta) -if (params.dimsum) { +if (params.fitness) { // MERGE_COUNTS.out.merged_counts shape: tuple( val([sample:'GID1A']), path("counts_merged.tsv") ) FIND_SYNONYMOUS_MUTATION( MERGE_COUNTS.out.merged_counts, // tuple(val(sample), path counts_merged.tsv) @@ -437,12 +438,12 @@ if (params.dimsum) { } -// --- DiMSum execution (only when --dimsum) --- -if (params.dimsum) { +// --- DiMSum execution (only when --fitness) --- +if (params.fitness) { // Shapes: // MERGE_COUNTS.out.merged_counts -> tuple(val([sample:'GID1A']), path('counts_merged.tsv')) // FIND_SYNONYMOUS_MUTATION.out.synonymous_wt -> tuple(val([sample:'GID1A']), path('synonymous_wt.txt')) - // EXPDESIGN_DIMSUM.out.experimental_design -> path('experimentalDesign.tsv') (singleton) + // EXPDESIGN_FITNESS.out.experimental_design -> path('experimentalDesign.tsv') (singleton) // 1) Key counts and WT by biological sample name to align them robustly def ch_counts_keyed_d = MERGE_COUNTS.out.merged_counts @@ -456,16 +457,19 @@ if (params.dimsum) { .map { key, smp, counts, wt -> tuple(smp, counts, wt) } // 3) Broadcast experimental design (singleton) to each sample triple - def ch_exp_for_each_d = EXPDESIGN_DIMSUM.out.experimental_design + def ch_exp_for_each_d = EXPDESIGN_FITNESS.out.experimental_design .combine(ch_counts_wt_d) .map { it[0] } - // 4) Final aligned channels for RUN_DIMSUM + // 4) Final aligned channels for FITNESS and RUN_DIMSUM def ch_run_counts_d = ch_counts_wt_d.map { smp, counts, wt -> tuple(smp, counts) } // matches: tuple val(sample), path(counts_merged) def ch_run_wt_d = ch_counts_wt_d.map { smp, counts, wt -> wt } // matches: path(wt_txt) def ch_run_exp_d = ch_exp_for_each_d // matches: path(exp_design) +} + +// --- DiMSum execution (only when --dimsum) --- +if (params.dimsum) { - // 5) Launch RUN_DIMSUM( ch_run_counts_d, // tuple val(sample), path(counts_merged) ch_run_wt_d, // path wt_txt From 89161ebd7c0d1a4ca92b9d0be33865cff786461b Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 <84199433+BenjaminWehnert1008@users.noreply.github.com> Date: Thu, 30 Oct 2025 14:50:42 +0100 Subject: [PATCH 274/317] Update deepmutscan.nf --- workflows/deepmutscan.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index 3ea467d..378cc1e 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -438,7 +438,7 @@ if (params.fitness) { } -// --- DiMSum execution (only when --fitness) --- +// --- fitness calculation (only when --fitness true) --- if (params.fitness) { // Shapes: // MERGE_COUNTS.out.merged_counts -> tuple(val([sample:'GID1A']), path('counts_merged.tsv')) From a3d999d5f62a312e16004f28b04c2c2fcf1314eb Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Thu, 30 Oct 2025 16:34:30 +0100 Subject: [PATCH 275/317] new default fitness calculation --- conf/modules.config | 10 +- .../dmsanalysis/bin/fitness_calculation.R | 301 ++++++++++++++++++ modules/local/dmsanalysis/bin/merge_counts.R | 4 - modules/local/fitness/fitness_calculation.nf | 40 +++ workflows/deepmutscan.nf | 33 +- 5 files changed, 371 insertions(+), 17 deletions(-) create mode 100644 modules/local/dmsanalysis/bin/fitness_calculation.R create mode 100644 modules/local/fitness/fitness_calculation.nf diff --git a/conf/modules.config b/conf/modules.config index db6ed04..dfc3c33 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -148,9 +148,17 @@ process { ] } + withName: 'FITNESS_CALCULATION' { + publishDir = [ + path: "${params.outdir}/fitness/output", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + withName: 'RUN_DIMSUM' { publishDir = [ - path: "${params.outdir}/DiMSum/output", + path: "${params.outdir}/DiMSum", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] diff --git a/modules/local/dmsanalysis/bin/fitness_calculation.R b/modules/local/dmsanalysis/bin/fitness_calculation.R new file mode 100644 index 0000000..b51e92c --- /dev/null +++ b/modules/local/dmsanalysis/bin/fitness_calculation.R @@ -0,0 +1,301 @@ +## default fitness estimation for nf-core/deepmutscan +## 27.10.2025 +## maximilian.stammnitz@crg.eu + +## 0. Libraries ## +################## + +suppressPackageStartupMessages({ + library(Biostrings) +}) + +## --- Helper functions --- + +# calculate nt hamming distances from the specified WT +compute_nt_hamming <- function(merged.counts, wt.seq) { + merged.counts <- cbind("nt_ham" = rep(NA, nrow(merged.counts)), merged.counts) + for (i in 1:nrow(merged.counts)){ + tmp.wt <- strsplit(as.character(wt.seq), "")[[1]] + tmp.mut <- strsplit(as.character(merged.counts$nt_seq[i]), "")[[1]] + if(length(which(tmp.mut != tmp.wt)) == 0){ + merged.counts$nt_ham[i] <- 0 + rm(tmp.mut, tmp.wt) + next + }else{ + merged.counts$nt_ham[i] <- length(which(tmp.mut != tmp.wt)) + rm(tmp.mut, tmp.wt) + next + } + } + merged.counts +} + +# translate sequences and add aa_seq +add_aa_seq <- function(merged.counts) { + merged.counts <- cbind("aa_seq" = as.character(translate(DNAStringSet(merged.counts$nt_seq))), merged.counts) + merged.counts +} + +# calculate AA hamming distances from the WT +compute_aa_hamming <- function(merged.counts, wt.seq.aa) { + merged.counts <- cbind("aa_ham" = rep(NA, nrow(merged.counts)), merged.counts) + for (i in 1:nrow(merged.counts)){ + tmp.wt <- strsplit(as.character(wt.seq.aa), "")[[1]] + tmp.mut <- strsplit(as.character(merged.counts$aa_seq[i]), "")[[1]] + if(length(which(tmp.mut != tmp.wt)) == 0){ + merged.counts$aa_ham[i] <- 0 + rm(tmp.mut, tmp.wt) + next + }else{ + merged.counts$aa_ham[i] <- length(which(tmp.mut != tmp.wt)) + rm(tmp.mut, tmp.wt) + next + } + } + merged.counts +} + +# name the mutations +name_mutations <- function(merged.counts, wt.seq.aa) { + merged.counts <- cbind("wt aa" = rep(NA, nrow(merged.counts)), + "pos" = rep(NA, nrow(merged.counts)), + "mut aa" = rep(NA, nrow(merged.counts)), merged.counts) + for (i in 1:nrow(merged.counts)){ + if(merged.counts$aa_ham[i] == 0){ + next + }else{ + tmp.wt <- strsplit(as.character(wt.seq.aa), "")[[1]] + tmp.mut <- strsplit(as.character(merged.counts$aa_seq[i]), "")[[1]] + merged.counts$pos[i] <- which(tmp.mut != tmp.wt) + merged.counts$`wt aa`[i] <- tmp.wt[merged.counts$pos[i]] + merged.counts$`mut aa`[i] <- tmp.mut[merged.counts$pos[i]] + rm(tmp.mut, tmp.wt) + } + } + merged.counts +} + +# find stops, WT and WT; aggregate counts of variants which are identical on the aa (but not nt) level +aggregate_by_aa <- function(merged.counts) { + ## find stops, WT and WT + merged.counts <- cbind(merged.counts, + "wt" = rep(NA, nrow(merged.counts)), + "stop" = rep(NA, nrow(merged.counts))) + merged.counts$wt[which(merged.counts$nt_ham == 0)] <- TRUE + merged.counts$stop[which(merged.counts$`mut aa` == "*")] <- TRUE + + ## aggregate counts of variants which are identical on the aa (but not nt) level + ## exception: wildtype ones + ## thereby shrinking the matrix + uniq.aa.vars <- unique(merged.counts$aa_seq) + uniq.aa.vars <- uniq.aa.vars[-which(uniq.aa.vars == merged.counts$aa_seq[which(merged.counts$wt == TRUE)])] + for(i in 1:length(uniq.aa.vars)){ + tmp.aa_seq <- uniq.aa.vars[i] + hits <- which(as.character(merged.counts$aa_seq) == tmp.aa_seq) + if(length(hits) == 1){ + rm(tmp.aa_seq, hits) + next + }else{ + for(j in grep("input|output", colnames(merged.counts))){ + merged.counts[hits[1],j] <- sum(merged.counts[hits,j], na.rm = TRUE) + } + merged.counts[hits[1], "nt_seq"] <- paste(merged.counts[hits, "nt_seq"], collapse = ", ") + merged.counts <- merged.counts[-hits[-1],] + rm(tmp.aa_seq, hits) + next + } + } + merged.counts +} + +# 3. Raw fitness calculations ## +calc_raw_fitness <- function(merged.counts, exp.design) { + ## how many fitness replicates are there + reps <- length(unique(exp.design$experiment_replicate)) + for (i in 1:reps){ + merged.counts <- cbind(merged.counts, rep(NA, nrow(merged.counts))) + colnames(merged.counts)[ncol(merged.counts)] <- paste0("raw_fitness_rep", i) + } + + ## calculate raw fitness of all variants vs. WT variant + for (i in 1:reps){ + + ### collect counts + tmp.input.counts <- merged.counts[,paste0("input", i)] + tmp.output.counts <- merged.counts[,paste0("output", i)] + + ### add pseudo-count to zero-outputs (if the corresponding input count is non-zero) + tmp.output.counts[which(tmp.output.counts == 0 & tmp.input.counts != 0)] <- 1 + + ### take logs + tmp.wt.log.ratio <- log(tmp.output.counts[which(merged.counts$wt == TRUE)] / + tmp.input.counts[which(merged.counts$wt == TRUE)]) + tmp.fitness <- log(tmp.output.counts / + tmp.input.counts) - tmp.wt.log.ratio + + ### uncertain values to NA + tmp.fitness[which(is.na(tmp.fitness) == TRUE)] <- NA + tmp.fitness[which(tmp.fitness == "Inf")] <- NA + + ### add to table + merged.counts[,c(ncol(merged.counts) - reps + i)] <- tmp.fitness + + ### clean up + rm(tmp.fitness, tmp.wt.log.ratio, tmp.output.counts, tmp.input.counts) + } + + list(merged.counts = merged.counts, reps = reps) +} + +# 4. Fitness and error refinements ## +rescale_and_summarize <- function(merged.counts, reps) { + ## center the raw fitness distributions on 0 (median of wildtype synonymous) and -1 (median of stops) + for (i in 1:reps){ + + merged.counts <- cbind(merged.counts, rep(NA, nrow(merged.counts))) + colnames(merged.counts)[ncol(merged.counts)] <- paste0("rescaled_fitness_rep", i) + + ### fetch the key counts + tmp.wt.fitness <- merged.counts[which(merged.counts$aa_ham == 0),ncol(merged.counts) - reps] + tmp.stop.fitness <- merged.counts[which(merged.counts$stop == TRUE),ncol(merged.counts) - reps] + + ### rescale + tmp.wt.fitness.med <- median(tmp.wt.fitness, na.rm = TRUE) + tmp.stop.fitness.med <- median(tmp.stop.fitness, na.rm = TRUE) + if(tmp.stop.fitness.med >= tmp.wt.fitness.med){ + + tmp.wt.fitness.mean <- mean(tmp.wt.fitness, na.rm = TRUE) + tmp.stop.fitness.mean <- mean(tmp.stop.fitness, na.rm = TRUE) + lm.rescale <- lm(c(0, -1) ~ c(tmp.wt.fitness.mean, tmp.stop.fitness.mean)) + merged.counts[,ncol(merged.counts)] <- merged.counts[,ncol(merged.counts) - reps] * lm.rescale$coefficients[[2]] + lm.rescale$coefficients[[1]] + rm(tmp.wt.fitness, tmp.stop.fitness, + tmp.wt.fitness.mean, tmp.stop.fitness.mean, + tmp.wt.fitness.med, tmp.stop.fitness.med, lm.rescale) + next + + }else{ + + lm.rescale <- lm(c(0, -1) ~ c(tmp.wt.fitness.med, tmp.stop.fitness.med)) + merged.counts[,ncol(merged.counts)] <- merged.counts[,ncol(merged.counts) - reps] * lm.rescale$coefficients[[2]] + lm.rescale$coefficients[[1]] + rm(tmp.wt.fitness, tmp.stop.fitness, + tmp.wt.fitness.med, tmp.stop.fitness.med, lm.rescale) + next + + } + } + + ## calculate fitness mean and standard deviation across replicates + merged.counts <- cbind(merged.counts, + "mean fitness" = rep(NA, nrow(merged.counts)), + "fitness sd" = rep(NA, nrow(merged.counts))) + + if(reps == 1){ + + merged.counts$`mean fitness` <- merged.counts[,ncol(merged.counts) - 2] + + }else if(reps > 1){ + + merged.counts$`mean fitness` <- apply(merged.counts[,c(ncol(merged.counts) - 2*reps + 1, ncol(merged.counts) - reps)], + 1, + mean, + na.rm = TRUE) + merged.counts$`fitness sd` <- apply(merged.counts[,c(ncol(merged.counts) - 2*reps + 1, ncol(merged.counts) - reps)], + 1, + sd, + na.rm = TRUE) + + } + + merged.counts +} + +## --- Main function --- + +#' Run default fitness estimation with configurable I/O paths +#' +#' @param counts_path Path to counts_merged.tsv +#' @param design_path Path to experimentalDesign.tsv +#' @param wt_seq_path Path to synonymous_wt.txt (single line DNA sequence) +#' @param output_path Path to write fitness_estimation.tsv +#' +#' @return Invisibly returns the final data.frame; writes the output to output_path. +run_fitness_estimation <- function(counts_path, + design_path, + wt_seq_path, + output_path) { + ## 1. Import key files ## + ######################### + + merged.counts <- read.table(counts_path, sep = "\t", header = TRUE, check.names = FALSE) + exp.design <- read.table(design_path, sep = "\t", header = TRUE, check.names = FALSE) + wt.seq <- DNAString(as.character(read.table(wt_seq_path))) + wt.seq.aa <- translate(wt.seq) + + ## 2. Pre-processing the count table ## + ####################################### + + ## calculate nt hamming distances from the specified WT + merged.counts <- compute_nt_hamming(merged.counts, wt.seq) + + ## translate sequences + merged.counts <- add_aa_seq(merged.counts) + + ## calculate AA hamming distances from the WT + merged.counts <- compute_aa_hamming(merged.counts, wt.seq.aa) + + ## name the mutations + merged.counts <- name_mutations(merged.counts, wt.seq.aa) + + ## find stops, WT and WT; aggregate AA-identical variants (except WT) + merged.counts <- aggregate_by_aa(merged.counts) + + ## 3. Raw fitness calculations ## + ################################# + fitness_res <- calc_raw_fitness(merged.counts, exp.design) + merged.counts <- fitness_res$merged.counts + reps <- fitness_res$reps + + ## 4. Fitness and error refinements ## + ###################################### + merged.counts <- rescale_and_summarize(merged.counts, reps) + + ## clean up + rm(reps) + + ## export + write.table(merged.counts, output_path, + col.names = TRUE, row.names = FALSE, quote = FALSE, sep = "\t", na = "") + + invisible(merged.counts) +} + + +## 5. Version ## +################ + +# sessionInfo() +# R version 4.5.1 (2025-06-13) +# Platform: aarch64-apple-darwin20 +# Running under: macOS Sonoma 14.6.1 +# +# Matrix products: default +# BLAS: /System/Library/Frameworks/Accelerate.framework/Versions/A/Frameworks/vecLib.framework/Versions/A/libBLAS.dylib +# LAPACK: /Library/Frameworks/R.framework/Versions/4.5-arm64/Resources/lib/libRlapack.dylib; LAPACK version 3.12.1 +# +# locale: +# [1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8 +# +# time zone: Europe/Madrid +# tzcode source: internal +# +# attached base packages: +# [1] stats4 stats graphics grDevices utils datasets methods base +# +# other attached packages: +# [1] Biostrings_2.76.0 GenomeInfoDb_1.44.2 XVector_0.48.0 IRanges_2.42.0 S4Vectors_0.46.0 +# [6] BiocGenerics_0.54.0 generics_0.1.4 +# +# loaded via a namespace (and not attached): +# [1] httr_1.4.7 compiler_4.5.1 R6_2.6.1 tools_4.5.1 +# [5] GenomeInfoDbData_1.2.14 rstudioapi_0.17.1 crayon_1.5.3 UCSC.utils_1.4.0 +# [9] jsonlite_2.0.0 \ No newline at end of file diff --git a/modules/local/dmsanalysis/bin/merge_counts.R b/modules/local/dmsanalysis/bin/merge_counts.R index db8d3d0..cfe5275 100644 --- a/modules/local/dmsanalysis/bin/merge_counts.R +++ b/modules/local/dmsanalysis/bin/merge_counts.R @@ -2,10 +2,6 @@ # Merges DiMSum-ready count tables into a single matrix with columns: # nt_seq, input1..inputN, output1..outputM -suppressMessages({ - # keep base R; if you prefer data.table, we can swap in later -}) - # ---- Core function ---- merge_dimsum_counts <- function(input_paths, output_paths, out_path = "counts.tsv") { # input_paths, output_paths: character vectors of file paths diff --git a/modules/local/fitness/fitness_calculation.nf b/modules/local/fitness/fitness_calculation.nf new file mode 100644 index 0000000..2495f05 --- /dev/null +++ b/modules/local/fitness/fitness_calculation.nf @@ -0,0 +1,40 @@ +process FITNESS_CALCULATION { + tag { sample.sample } + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" + + input: + tuple val(sample), path(counts_merged) + path(exp_design) + path(syn_wt_txt) + path script // fitness_calculation.R + + output: + tuple val(sample), path("fitness_estimation.tsv"), emit: fitness_estimation + path "versions.yml", emit: versions + + script: + """ + set -euo pipefail + + R_version=\$(R --version | head -n 1 | sed 's/^R version //') + + Rscript -e "source('$script'); run_fitness_estimation('$counts_merged', '$exp_design', '$syn_wt_txt', 'fitness_estimation.tsv')" + + cat > versions.yml < versions.yml + echo " stub-version: 0.0.0" >> versions.yml + """ +} diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index 3ea467d..3968970 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -23,6 +23,7 @@ include { GATK_GATKTOFITNESS } from '../modules/local/gatk/gatktofitnes include { MERGE_COUNTS } from '../modules/local/fitness/merge_counts' include { EXPDESIGN_FITNESS } from '../modules/local/fitness/fitness_experimental_design' include { FIND_SYNONYMOUS_MUTATION } from '../modules/local/fitness/find_synonymous_mutation' +include { FITNESS_CALCULATION } from '../modules/local/fitness/fitness_calculation' include { RUN_DIMSUM } from '../modules/local/dimsum/run_dimsum' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -36,14 +37,14 @@ include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_deep */ // Params defaults -params.min_counts = params.min_counts ?: 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library -params.sliding_window_size = params.sliding_window_size ?: 10 // sliding window size to flatten graphs in plots (e.g. GLOBAL_POS_BIASES_COUNTS function) -params.aimed_cov = params.aimed_cov ?: 100 // aimed coverage (assuming equal spread) to visualize threshold in plots -params.run_seqdepth = params.run_seqdepth ?: false // creating seqdepth simulation plot, is computationally quite heavy. per default disabled. -params.fitness = params.fitness ?: false // run basic fitness calculation from selection input & output samples -params.dimsum = params.dimsum ?: false // run DiMSum for fitness/functionality scores from selection input & output samples +params.min_counts = 3 // minimum counts for variant to be recognized. All variants>custom<< this variable has to be path to .txt with custom library +params.sliding_window_size = 10 // sliding window size to flatten graphs in plots (e.g. GLOBAL_POS_BIASES_COUNTS function) +params.aimed_cov = 100 // aimed coverage (assuming equal spread) to visualize threshold in plots +params.run_seqdepth = false // creating seqdepth simulation plot, is computationally quite heavy. per default disabled. +params.fitness = false // run basic fitness calculation from selection input & output samples +params.dimsum = false // run DiMSum for fitness/functionality scores from selection input & output samples // Define fasta file as channel (e.g. for BWA index) @@ -118,6 +119,7 @@ R("modules/local/dmsanalysis/bin/prepare_gatk_data_for_fitness_heatmap.R").set { R("modules/local/dmsanalysis/bin/process_raw_gatk.R").set { process_raw_gatk_script_ch } R("modules/local/dmsanalysis/bin/merge_counts.R").set { merge_counts_script_ch } R("modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R").set { exp_design_ch } +R("modules/local/dmsanalysis/bin/fitness_calculation.R").set { fitness_calculation_script_ch } R("modules/local/dmsanalysis/bin/find_syn_mutation.R").set { syn_mut_ch } @@ -423,7 +425,7 @@ logdiff_scriptN = fanoutTo(library_completed_variantCounts_ch, lo } -// --- Synonymous WT selection (runs only when --dimsum) --- +// --- Synonymous WT selection (runs only when --fitness) --- // Strip meta once: keep only the fasta path ch_fasta.map { it[1] }.set { ch_fasta_path } // path(/…/GID1A.fasta) @@ -438,7 +440,7 @@ if (params.fitness) { } -// --- DiMSum execution (only when --fitness) --- +// --- Fitness execution (only when --fitness) --- if (params.fitness) { // Shapes: // MERGE_COUNTS.out.merged_counts -> tuple(val([sample:'GID1A']), path('counts_merged.tsv')) @@ -461,10 +463,17 @@ if (params.fitness) { .combine(ch_counts_wt_d) .map { it[0] } - // 4) Final aligned channels for FITNESS and RUN_DIMSUM + // 4) Final aligned channels for FITNESS_CALCULATION and RUN_DIMSUM def ch_run_counts_d = ch_counts_wt_d.map { smp, counts, wt -> tuple(smp, counts) } // matches: tuple val(sample), path(counts_merged) def ch_run_wt_d = ch_counts_wt_d.map { smp, counts, wt -> wt } // matches: path(wt_txt) def ch_run_exp_d = ch_exp_for_each_d // matches: path(exp_design) + + FITNESS_CALCULATION( + ch_run_counts_d, // tuple val(sample), path(counts_merged) + ch_run_exp_d, // path experimentalDesign.tsv + ch_run_wt_d, // path syn_wt_txt + fitness_calculation_script_ch + ) } // --- DiMSum execution (only when --dimsum) --- @@ -472,7 +481,7 @@ if (params.dimsum) { RUN_DIMSUM( ch_run_counts_d, // tuple val(sample), path(counts_merged) - ch_run_wt_d, // path wt_txt + ch_run_wt_d, // path syn_wt_txt ch_run_exp_d // path experimentalDesign.tsv ) } From de284cb2ce6b716b13135b0383523d115fa6d4ef Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Thu, 30 Oct 2025 17:11:09 +0100 Subject: [PATCH 276/317] make dimsum conditional to --fitness true --- workflows/deepmutscan.nf | 20 ++++++++------------ 1 file changed, 8 insertions(+), 12 deletions(-) diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index 1d402e7..065a5e0 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -474,24 +474,20 @@ if (params.fitness) { ch_run_wt_d, // path syn_wt_txt fitness_calculation_script_ch ) -} - -// --- DiMSum execution (only when --dimsum) --- -if (params.dimsum) { - RUN_DIMSUM( - ch_run_counts_d, // tuple val(sample), path(counts_merged) - ch_run_wt_d, // path syn_wt_txt - ch_run_exp_d // path experimentalDesign.tsv - ) +// --- DiMSum execution (only when --fitness true & --dimsum true) --- + if (params.dimsum) { + RUN_DIMSUM( + ch_run_counts_d, // tuple val(sample), path(counts_merged) + ch_run_wt_d, // path syn_wt_txt + ch_run_exp_d // path experimentalDesign.tsv + ) + } } - - - emit: multiqc_report = MULTIQC.out.report.toList() // channel: /path/to/multiqc_report.html versions = ch_versions // channel: [ path(versions.yml) ] From 3709de165b6bf03acd2c2cdbd17f9ed85921aadf Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Thu, 30 Oct 2025 17:25:55 +0100 Subject: [PATCH 277/317] automated compatibility warning when using DiMSum --- workflows/deepmutscan.nf | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index 065a5e0..9666bc5 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -439,6 +439,14 @@ if (params.fitness) { ) } +// Warning on compatibilities of DiMSum if --dimsum true. +if (params.dimsum) { + log.warn(""" + '--dimsum true' only works together with '--fitness true' + and is currently (30 Oct 2025) NOT supported on ARM processors. + Use AMD/x86_64 systems for DiMSum execution. + """) +} // --- fitness calculation (only when --fitness true) --- if (params.fitness) { From d1fcb3458196e48b5c7af794a111b3c335ea8358 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Fri, 31 Oct 2025 13:38:04 +0100 Subject: [PATCH 278/317] added base R fitness correlation plots --- conf/modules.config | 10 +- modules/local/dmsanalysis/bin/fitness_QC.R | 99 +++++++++++++++++++ modules/local/fitness/fitness_calculation.nf | 40 -------- modules/local/fitness/fitness_standard.nf | 88 +++++++++++++++++ .../local/{dimsum => fitness}/run_dimsum.nf | 0 workflows/deepmutscan.nf | 11 ++- 6 files changed, 205 insertions(+), 43 deletions(-) create mode 100644 modules/local/dmsanalysis/bin/fitness_QC.R delete mode 100644 modules/local/fitness/fitness_calculation.nf create mode 100644 modules/local/fitness/fitness_standard.nf rename modules/local/{dimsum => fitness}/run_dimsum.nf (100%) diff --git a/conf/modules.config b/conf/modules.config index dfc3c33..476ef45 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -106,7 +106,7 @@ process { withName: /.*VISUALIZATION_.*/ { publishDir = [ - path: { "${params.outdir}/final_plots" }, // e.g. results/final_plots + path: { "${params.outdir}/library_QC" }, // e.g. results/library_QC mode: 'copy', overwrite: false, saveAs: { fn -> @@ -156,6 +156,14 @@ process { ] } + withName: 'FITNESS_QC' { + publishDir = [ + path: "${params.outdir}/fitness/output", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + withName: 'RUN_DIMSUM' { publishDir = [ path: "${params.outdir}/DiMSum", diff --git a/modules/local/dmsanalysis/bin/fitness_QC.R b/modules/local/dmsanalysis/bin/fitness_QC.R new file mode 100644 index 0000000..4118ccd --- /dev/null +++ b/modules/local/dmsanalysis/bin/fitness_QC.R @@ -0,0 +1,99 @@ +## fitness QC plots for nf-core/deepmutscan +## 28.10.2025 +## maximilian.stammnitz@crg.eu + +## lower panels: scatter + x=y (log-log version for counts) +panel_xy_abline_counts <- function(x, y, ...) { + op <- par("xpd"); on.exit(par(xpd = op), add = TRUE) + par(xpd = FALSE) + points(x, y, pch = 16, cex = 0.1, ...) + abline(a = 0, b = 1, lty = 2, col = "grey50") +} + +## upper panels: Pearson r (log-log-transformed) +panel_cor_counts <- function(x, y, digits = 2, prefix = "r = ", cex.text = 1.4, ...) { + r <- suppressWarnings(cor(log(x), log(y), use = "pairwise.complete.obs", method = "pearson")) + lab <- if (is.finite(r)) bquote(italic(r) == .(round(r, digits))) else bquote(italic(r) == NA) + + ## Save/restore full graphics state we touch + op <- par(c("usr", "xpd", "xlog", "ylog")) + on.exit(par(op), add = TRUE) + + ## Draw in normalized 0..1 panel coords with logs OFF so text is visible + par(xlog = FALSE, ylog = FALSE, xpd = FALSE, usr = c(0, 1, 0, 1)) + text(0.5, 0.5, labels = lab, cex = cex.text, font = 1, col = "black") +} + +## lower panels: scatter + x=y (linear version for fitness) +panel_xy_abline_fitness <- function(x, y, ...) { + op <- par("xpd"); on.exit(par(xpd = op), add = TRUE) + par(xpd = FALSE) + points(x, y, pch = 16, cex = 0.5, ...) + abline(a = 0, b = 1, lty = 2, col = "grey50") +} + +## upper panels: Pearson r (linear) +panel_cor_fitness <- function(x, y, digits = 2, prefix = "r = ", cex.text = 1.4, ...) { + r <- suppressWarnings(cor(x, y, use = "pairwise.complete.obs", method = "pearson")) + lab <- if (is.finite(r)) bquote(italic(r) == .(round(r, digits))) else bquote(italic(r) == NA) + + ## Save/restore full graphics state we touch + op <- par(c("usr", "xpd", "xlog", "ylog")) + on.exit(par(op), add = TRUE) + + ## Draw in normalized 0..1 panel coords with logs OFF so text is visible + par(xlog = FALSE, ylog = FALSE, xpd = FALSE, usr = c(0, 1, 0, 1)) + text(0.5, 0.5, labels = lab, cex = cex.text, font = 1, col = "black") +} + +#' Plot input/output count correlations and fitness replicate correlations +#' +#' @param fitness_table_path Path to the input table (fitness_estimation.tsv) +#' @param out_counts_corr_pdf Path to write the counts correlation PDF +#' @param out_fitness_corr_pdf Path to write the fitness correlation PDF +#' +#' @return Invisibly returns TRUE; writes the two PDFs. +run_fitness_plots <- function(fitness_table_path, + out_counts_corr_pdf, + out_fitness_corr_pdf) { + + merged.counts.fitness <- read.table(fitness_table_path, sep = "\t", header = TRUE, check.names = FALSE) + + ## identify the right samples + inputs <- grep("input", colnames(merged.counts.fitness)) + outputs <- grep("output", colnames(merged.counts.fitness)) + + ## 5. Plot input vs. output counts ## + ##################################### + pdf(out_counts_corr_pdf, height = 9, width = 14) + pairs(merged.counts.fitness[, c(inputs, outputs)] + 1, ## use a pseudo-count + lower.panel = panel_xy_abline_counts, + upper.panel = panel_cor_counts, + cex.text = 2, + log = "xy") + dev.off() + + ## 6. Plot fitness correlations ## + ################################## + fitness.repl <- grep("rescaled_fitness", colnames(merged.counts.fitness)) + + if (length(fitness.repl) > 1) { + pdf(out_fitness_corr_pdf, height = 9, width = 14) + pairs(merged.counts.fitness[, fitness.repl], + lower.panel = panel_xy_abline_fitness, + upper.panel = panel_cor_fitness, + cex.text = 2, + xlim = c(-3, 1), + ylim = c(-3, 1)) + dev.off() + } else { + ## If only one (or zero) rescaled_fitness columns exist, still create an empty placeholder + ## so Nextflow finds the declared output. + pdf(out_fitness_corr_pdf, height = 9, width = 14) + plot.new() + title("No replicate fitness columns found (need ≥2 'rescaled_fitness...')\nCreated placeholder PDF.") + dev.off() + } + + invisible(TRUE) +} diff --git a/modules/local/fitness/fitness_calculation.nf b/modules/local/fitness/fitness_calculation.nf deleted file mode 100644 index 2495f05..0000000 --- a/modules/local/fitness/fitness_calculation.nf +++ /dev/null @@ -1,40 +0,0 @@ -process FITNESS_CALCULATION { - tag { sample.sample } - label 'process_single' - - conda "${moduleDir}/environment.yml" - container "${ workflow.containerEngine == 'singularity' - ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' - : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" - - input: - tuple val(sample), path(counts_merged) - path(exp_design) - path(syn_wt_txt) - path script // fitness_calculation.R - - output: - tuple val(sample), path("fitness_estimation.tsv"), emit: fitness_estimation - path "versions.yml", emit: versions - - script: - """ - set -euo pipefail - - R_version=\$(R --version | head -n 1 | sed 's/^R version //') - - Rscript -e "source('$script'); run_fitness_estimation('$counts_merged', '$exp_design', '$syn_wt_txt', 'fitness_estimation.tsv')" - - cat > versions.yml < versions.yml - echo " stub-version: 0.0.0" >> versions.yml - """ -} diff --git a/modules/local/fitness/fitness_standard.nf b/modules/local/fitness/fitness_standard.nf new file mode 100644 index 0000000..c222833 --- /dev/null +++ b/modules/local/fitness/fitness_standard.nf @@ -0,0 +1,88 @@ +process FITNESS_CALCULATION { + tag { sample.sample } + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" + + input: + tuple val(sample), path(counts_merged) + path(exp_design) + path(syn_wt_txt) + path script // fitness_calculation.R + + output: + tuple val(sample), path("fitness_estimation.tsv"), emit: fitness_estimation + path "versions.yml", emit: versions + + script: + """ + set -euo pipefail + + R_version=\$(R --version | head -n 1 | sed 's/^R version //') + + Rscript -e "source('$script'); run_fitness_estimation('$counts_merged', '$exp_design', '$syn_wt_txt', 'fitness_estimation.tsv')" + + cat > versions.yml < versions.yml + echo " stub-version: 0.0.0" >> versions.yml + """ +} + + + +process FITNESS_QC { + tag { sample.sample } + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" + + input: + tuple val(sample), path(fitness_estimation_tsv) // from FITNESS_CALCULATION + path script // fitness_plots.R + + output: + tuple val(sample), path("fitness_estimation_count_correlation.pdf"), emit: counts_corr_pdf + tuple val(sample), path("fitness_estimation_fitness_correlation.pdf"), emit: fitness_corr_pdf + path "versions.yml", emit: versions + + script: + """ + set -euo pipefail + + R_version=\$(R --version | head -n 1 | sed 's/^R version //') + + Rscript -e "source('$script'); run_fitness_plots( + '$fitness_estimation_tsv', + 'fitness_estimation_count_correlation.pdf', + 'fitness_estimation_fitness_correlation.pdf')" + + cat > versions.yml < versions.yml <<'EOF' + FITNESS_PLOTS: + stub-version: "0.0.0" + EOF + """ +} diff --git a/modules/local/dimsum/run_dimsum.nf b/modules/local/fitness/run_dimsum.nf similarity index 100% rename from modules/local/dimsum/run_dimsum.nf rename to modules/local/fitness/run_dimsum.nf diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index 9666bc5..1ae7d40 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -23,8 +23,9 @@ include { GATK_GATKTOFITNESS } from '../modules/local/gatk/gatktofitnes include { MERGE_COUNTS } from '../modules/local/fitness/merge_counts' include { EXPDESIGN_FITNESS } from '../modules/local/fitness/fitness_experimental_design' include { FIND_SYNONYMOUS_MUTATION } from '../modules/local/fitness/find_synonymous_mutation' -include { FITNESS_CALCULATION } from '../modules/local/fitness/fitness_calculation' -include { RUN_DIMSUM } from '../modules/local/dimsum/run_dimsum' +include { FITNESS_CALCULATION } from '../modules/local/fitness/fitness_standard' +include { FITNESS_QC } from '../modules/local/fitness/fitness_standard' +include { RUN_DIMSUM } from '../modules/local/fitness/run_dimsum' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -120,6 +121,7 @@ R("modules/local/dmsanalysis/bin/process_raw_gatk.R").set { process_raw_gatk_scr R("modules/local/dmsanalysis/bin/merge_counts.R").set { merge_counts_script_ch } R("modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R").set { exp_design_ch } R("modules/local/dmsanalysis/bin/fitness_calculation.R").set { fitness_calculation_script_ch } +R("modules/local/dmsanalysis/bin/fitness_QC.R").set { fitness_QC_script_ch } R("modules/local/dmsanalysis/bin/find_syn_mutation.R").set { syn_mut_ch } @@ -483,6 +485,11 @@ if (params.fitness) { fitness_calculation_script_ch ) + FITNESS_QC( + FITNESS_CALCULATION.out.fitness_estimation, // tuple val(sample), path(fitness_estimation.tsv) + fitness_QC_script_ch + ) + // --- DiMSum execution (only when --fitness true & --dimsum true) --- if (params.dimsum) { RUN_DIMSUM( From 781963edbe74afc32861dd42a5a0484db210f92b Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Mon, 3 Nov 2025 11:57:21 +0100 Subject: [PATCH 279/317] fitness heatmap with correct amino acid order --- conf/modules.config | 12 +- .../local/dmsanalysis/bin/counts_heatmap.R | 13 +- .../dmsanalysis/bin/counts_per_cov_heatmap.R | 13 +- .../local/dmsanalysis/bin/fitness_heatmap.R | 386 +++++++++++------- modules/local/fitness/fitness_standard.nf | 45 ++ workflows/deepmutscan.nf | 8 + 6 files changed, 330 insertions(+), 147 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 476ef45..5f19e0e 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -118,7 +118,7 @@ process { withName: 'GATK_GATKTOFITNESS' { publishDir = [ - path: "${params.outdir}/DiMSum/single_counts", + path: "${params.outdir}/fitness/DiMSum/single_counts", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] @@ -164,9 +164,17 @@ process { ] } + withName: 'FITNESS_HEATMAP' { + publishDir = [ + path: "${params.outdir}/fitness/output", + mode: 'copy', + saveAs: { filename -> filename == 'versions.yml' ? null : filename } + ] + } + withName: 'RUN_DIMSUM' { publishDir = [ - path: "${params.outdir}/DiMSum", + path: "${params.outdir}/fitness/DiMSum", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] diff --git a/modules/local/dmsanalysis/bin/counts_heatmap.R b/modules/local/dmsanalysis/bin/counts_heatmap.R index dd37b2e..f685ef5 100644 --- a/modules/local/dmsanalysis/bin/counts_heatmap.R +++ b/modules/local/dmsanalysis/bin/counts_heatmap.R @@ -8,8 +8,9 @@ counts_heatmap <- function(input_csv_path, threshold = 3, output_pdf_path, img_f # Inner function to add padding to the last row, adding 21 amino acids per position pad_heatmap_data_long <- function(heatmap_data_long, min_non_na_value, num_positions_per_row = 75) { - all_amino_acids <- c("A", "C", "D", "E", "F", "G", "H", "I", "K", "L", - "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y", "*") + all_amino_acids <- c("G", "A", "V", "L", "M", "I", "F", + "Y", "W", "K", "R", "H", "D", "E", + "S", "T", "C", "N", "Q", "P", "*") max_position <- max(heatmap_data_long$position) num_missing_positions <- num_positions_per_row - (max_position %% num_positions_per_row) @@ -69,8 +70,12 @@ counts_heatmap <- function(input_csv_path, threshold = 3, output_pdf_path, img_f mutate(synonymous = mut_aa == wt_aa) # Definiere die korrekte Reihenfolge der Aminosäuren - amino_acid_order <- c("*", "A", "C", "D", "E", "F", "G", "H", "I", "K", - "L", "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y") + amino_acid_order <- c("G", "A", "V", "L", "M", "I", "F", + "Y", "W", "K", "R", "H", "D", "E", + "S", "T", "C", "N", "Q", "P", "*") + + heatmap_data_long <- heatmap_data_long %>% + mutate(mut_aa = factor(mut_aa, levels = amino_acid_order)) # Bearbeite heatmap_data_long und erstelle syn_positions gleichzeitig syn_positions <- heatmap_data_long %>% diff --git a/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R b/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R index 8af6b80..b51668e 100644 --- a/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R +++ b/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R @@ -8,8 +8,9 @@ counts_per_cov_heatmap <- function(input_csv_path, threshold = 3, output_pdf_pat # Inner function to add padding to the last row, adding 21 amino acids per position pad_heatmap_data_long <- function(heatmap_data_long, min_non_na_value, num_positions_per_row = 75) { - all_amino_acids <- c("A", "C", "D", "E", "F", "G", "H", "I", "K", "L", - "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y", "*") + all_amino_acids <- c("G", "A", "V", "L", "M", "I", "F", + "Y", "W", "K", "R", "H", "D", "E", + "S", "T", "C", "N", "Q", "P", "*") max_position <- max(heatmap_data_long$position) num_missing_positions <- num_positions_per_row - (max_position %% num_positions_per_row) @@ -69,8 +70,12 @@ counts_per_cov_heatmap <- function(input_csv_path, threshold = 3, output_pdf_pat mutate(synonymous = mut_aa == wt_aa) # Definiere die korrekte Reihenfolge der Aminosäuren - amino_acid_order <- c("*", "A", "C", "D", "E", "F", "G", "H", "I", "K", - "L", "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y") + amino_acid_order <- c("G", "A", "V", "L", "M", "I", "F", + "Y", "W", "K", "R", "H", "D", "E", + "S", "T", "C", "N", "Q", "P", "*") + + heatmap_data_long <- heatmap_data_long %>% + mutate(mut_aa = factor(mut_aa, levels = amino_acid_order)) # Bearbeite heatmap_data_long und erstelle syn_positions gleichzeitig syn_positions <- heatmap_data_long %>% diff --git a/modules/local/dmsanalysis/bin/fitness_heatmap.R b/modules/local/dmsanalysis/bin/fitness_heatmap.R index 09e7d57..348fa9f 100644 --- a/modules/local/dmsanalysis/bin/fitness_heatmap.R +++ b/modules/local/dmsanalysis/bin/fitness_heatmap.R @@ -1,160 +1,272 @@ -library(dplyr) -library(ggplot2) +suppressPackageStartupMessages({ + library(methods) + library(dplyr) + library(ggplot2) + library(grid) # for unit() +}) -fitness_heatmap <- function(input_csv_path, threshold = 3, output_pdf_path, img_format = "pdf") { +# ---------- helper functions ---------- +find_col <- function(df, candidates) { + norm <- function(x) gsub("[^a-z0-9]+", "_", tolower(x)) + nms <- colnames(df); nn <- norm(nms) + for (cand in candidates) { + hit <- which(nn == norm(cand)) + if (length(hit) == 1) return(nms[hit]) + } + stop(sprintf("Could not find any of columns: %s", paste(candidates, collapse = ", "))) +} - # Inner function to add padding to the last row, adding 21 amino acids per position - pad_heatmap_data_long <- function(heatmap_data_long, min_non_na_value, num_positions_per_row = 75) { - all_amino_acids <- c("A", "C", "D", "E", "F", "G", "H", "I", "K", "L", - "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y", "*") +get_rescaled_cols <- function(df) { + nms <- colnames(df) + hits <- grep("^rescaled[_ ]?fitness", nms, ignore.case = TRUE, value = TRUE) + if (!length(hits)) stop("No 'rescaled_fitness' columns found.") + idx <- suppressWarnings(as.integer(gsub(".*?([0-9]+)$", "\\1", hits))) + hits[order(is.na(idx), idx, hits)] +} - max_position <- max(heatmap_data_long$position) - num_missing_positions <- num_positions_per_row - (max_position %% num_positions_per_row) +# Find "mean fitness" column (if present) +find_mean_col <- function(df) { + nms <- colnames(df) + key <- tolower(gsub("[^a-z0-9]+", "_", nms)) + hit <- which(key == "mean_fitness") + if (length(hit) == 1) nms[hit] else NULL +} - if (num_missing_positions < num_positions_per_row) { - new_positions <- (max_position + 1):(max_position + num_missing_positions) +# NEW: read WT amino acid sequence from .txt file (single line) +read_wt_seq_aa_txt <- function(path) { + if (is.null(path)) stop("wt_seq_aa_txt_path must be provided.") + x <- readLines(path, warn = FALSE) + x <- x[nzchar(x)] + if (!length(x)) stop("WT AA TXT is empty.") + aa <- toupper(gsub("\\s+", "", x[which.max(nchar(x))])) + # Keep only valid amino acid letters (including stop '*') + aa <- gsub("[^ACDEFGHIKLMNPQRSTVWY*]", "", aa) + if (!nchar(aa)) stop("WT AA TXT contains no valid AA letters.") + aa +} - # Add all 21 amino acid variants for each new position - padding_data <- expand.grid( - mut_aa = all_amino_acids, # All possible amino acids - position = new_positions # New positions to be padded - ) +# Build full AA×position grid for positions 1..wt_len (from WT sequence), +# join fitness values; any missing combos stay NA (-> grey). +# Positions > wt_len are considered padded and will be white. +build_heatmap_long <- function(df, + wt_aa_col, + pos_col, + mut_aa_col, + fitness_col, + positions_per_row = 75, + wt_seq_aa, + fill_missing_as_zero = FALSE) { - # Set placeholder values for the added positions to the exact smallest non-NA value - padding_data$total_counts_per_cov <- min_non_na_value # Set to the smallest non-NA value - padding_data$wt_aa <- "Y" # Set wild-type amino acid to 'Y' - padding_data$wt_aa_pos <- paste0("Y", padding_data$position) # Create wt_aa_pos with correct positions - padding_data$row_group <- max(heatmap_data_long$row_group) # Set row group to the current last group + # authoritative WT length from provided sequence + letters <- strsplit(wt_seq_aa, "", fixed = TRUE)[[1]] + wt_len <- length(letters) - # Add the new padding rows to heatmap_data_long - heatmap_data_long <- dplyr::bind_rows(heatmap_data_long, padding_data) - } + # normalize data + df0 <- df %>% + transmute( + position = suppressWarnings(as.numeric(.data[[pos_col]])), + wt_aa_in = .data[[wt_aa_col]], + mut_aa = .data[[mut_aa_col]], + fitness = suppressWarnings(as.numeric(.data[[fitness_col]])) + ) %>% + filter(is.finite(position)) - return(heatmap_data_long) + # drop any rows that claim positions beyond WT length + if (nrow(df0) && any(df0$position > wt_len, na.rm = TRUE)) { + dropped <- sum(df0$position > wt_len, na.rm = TRUE) + warning(sprintf("Dropping %d row(s) with position > WT length (%d).", dropped, wt_len)) + df0 <- df0 %>% filter(position <= wt_len) } - # Load the CSV data - heatmap_data <- read.csv(input_csv_path) + # pad to next multiple of 75 (by rows) + rem <- wt_len %% positions_per_row + pad_need <- if (rem == 0) 0 else positions_per_row - rem + max_paded <- wt_len + pad_need - # Check if the necessary column exists in the data - if (!"fitness" %in% colnames(heatmap_data)) { - stop("The column 'fitness' is not found in the data.") + # full grid: positions 1..max_paded (so the tail exists), AA set of 21 + all_positions <- seq_len(max_paded) + all_amino_acids <- c("A","C","D","E","F","G","H","I","K","L", + "M","N","P","Q","R","S","T","V","W","Y","*") + + grid_df <- expand.grid(position = all_positions, + mut_aa = all_amino_acids, + KEEP.OUT.ATTRS = FALSE, stringsAsFactors = FALSE) %>% + mutate(is_padded = position > wt_len) + + # join fitness only for real positions (<= wt_len) + fit_df <- df0 %>% select(position, mut_aa, fitness) + d <- grid_df %>% + left_join(fit_df, by = c("position","mut_aa")) + + if (fill_missing_as_zero) { + d$fitness[is.na(d$fitness) & d$position <= wt_len] <- 0 } - # Create heatmap_data_long by selecting necessary columns - heatmap_data_long <- heatmap_data %>% - select(mut_aa, position, fitness, wt_aa) # Use 'fitness' - - # Find the smallest non-NA value in fitness column - min_non_na_value <- min(heatmap_data_long$fitness, na.rm = TRUE) - - # Group positions by rows (75 positions per row) and calculate row_group - heatmap_data_long <- heatmap_data_long %>% - mutate(row_group = ((position - 1) %/% 75) + 1) # Grouping positions into rows - - # Apply padding to add missing positions at the end of the last row, using the calculated min value - heatmap_data_long <- pad_heatmap_data_long(heatmap_data_long, min_non_na_value) - - # Convert positions to numeric, sort them, and create wt_aa_pos for the plot - heatmap_data_long <- heatmap_data_long %>% - mutate(position = as.numeric(position)) %>% # Ensure position is numeric - arrange(position) %>% # Sort by position - mutate(wt_aa_pos = factor(paste0(wt_aa, position), levels = unique(paste0(wt_aa, position)))) # Create sorted factor levels for wt_aa_pos - - # Add a column to identify synonymous mutations (where mut_aa == wt_aa) - heatmap_data_long <- heatmap_data_long %>% - mutate(synonymous = mut_aa == wt_aa) - - # Definiere die korrekte Reihenfolge der Aminosäuren - amino_acid_order <- c("*", "A", "C", "D", "E", "F", "G", "H", "I", "K", - "L", "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y") - - # Bearbeite heatmap_data_long und erstelle syn_positions gleichzeitig - syn_positions <- heatmap_data_long %>% - mutate(mut_aa = factor(mut_aa, levels = amino_acid_order), - # Berechne die x-Koordinate, die pro Gruppe immer von 1 bis 75 verläuft - x = as.numeric(factor(wt_aa_pos, levels = unique(wt_aa_pos))) - ((row_group - 1) * 75), - y = as.numeric(factor(mut_aa, levels = amino_acid_order))) %>% - filter(synonymous == TRUE) - - # Calculate the number of row groups and adjust plot height dynamically - num_row_groups <- max(heatmap_data_long$row_group) - plot_height <- num_row_groups * 4 - - # Set the limits for the color scale, ignoring NA (negative values are now NA) - min_count <- min(heatmap_data_long$fitness, na.rm = TRUE) - max_count <- max(heatmap_data_long$fitness, na.rm = TRUE) - max_position <- max(heatmap_data$position) - - # Create the heatmap plot with explicit handling for positions > max_position - heatmap_plot <- ggplot(heatmap_data_long, aes(x = wt_aa_pos, y = mut_aa, fill = fitness)) + + # authoritative WT AA per real position; tail gets placeholder 'Y' + wt_map <- tibble(position = seq_len(wt_len), wt_aa = letters) + d <- d %>% + left_join(wt_map, by = "position") %>% + mutate(wt_aa = ifelse(is.na(wt_aa) & position > wt_len, "Y", wt_aa)) + + # layout fields + d <- d %>% + mutate( + row_group = ((position - 1) %/% positions_per_row) + 1, + wt_aa_pos = paste0(wt_aa, position), + wt_aa_pos = factor(wt_aa_pos, levels = unique(wt_aa_pos)), + synonymous = mut_aa == wt_aa + ) + + # IMPORTANT: use WT length as the true end of the protein + d$max_pos <- wt_len + d +} + +syn_segments <- function(d, positions_per_row = 75) { + amino_order <- c("G", "A", "V", "L", "M", "I", "F", + "Y", "W", "K", "R", "H", "D", "E", + "S", "T", "C", "N", "Q", "P", "*") + d %>% + mutate( + mut_aa = factor(mut_aa, levels = amino_order), + x = as.numeric(factor(wt_aa_pos, levels = levels(wt_aa_pos))) - + ((row_group - 1) * positions_per_row), + y = as.numeric(factor(mut_aa, levels = amino_order)) + ) %>% + filter(synonymous, position <= max_pos) +} + +# Draw one solid white rectangle per row group covering the padded tail region +white_tail_rects <- function(d, positions_per_row = 75) { + wt_len <- unique(d$max_pos)[1] + if (!is.finite(wt_len)) return(dplyr::tibble()[0,]) + + # if perfectly divisible by 75, there is no tail to cover + if (wt_len %% positions_per_row == 0) return(dplyr::tibble()[0,]) + + # which facet (row group) contains the last real position? + last_group <- ((wt_len - 1) %/% positions_per_row) + 1 + last_local_idx <- ((wt_len - 1) %% positions_per_row) + 1 # 1..75 within the facet + + tibble::tibble( + row_group = last_group, + xmin = last_local_idx + 0.5 - 0.025, # tiny epsilon to avoid hairlines + xmax = positions_per_row + 0.5 + 0.025, + ymin = 0.5 - 0.025, + ymax = 21.5 + 0.025 + ) +} + +plot_heatmap <- function(d, title_text, positions_per_row = 75) { + amino_order <- c("G", "A", "V", "L", "M", "I", "F", + "Y", "W", "K", "R", "H", "D", "E", + "S", "T", "C", "N", "Q", "P", "*") + d <- d %>% mutate(mut_aa = factor(mut_aa, levels = amino_order)) + + min_f <- suppressWarnings(min(d$fitness, na.rm = TRUE)); if (!is.finite(min_f)) min_f <- 0 + max_f <- suppressWarnings(max(d$fitness, na.rm = TRUE)); if (!is.finite(max_f)) max_f <- 0 + max_orig_pos <- unique(d$max_pos)[1] + + syn <- syn_segments(d, positions_per_row) + rect <- white_tail_rects(d, positions_per_row) + + ggplot(d, aes(x = wt_aa_pos, y = mut_aa, fill = fitness)) + scale_fill_gradientn( - colours = c("#D73027", "#F0F0F0", "#4575B4"), # From low = red to high = blue - values = c(0, abs(min_count) / (abs(min_count) + max_count), 1), # Manually define proportions - na.value = "grey35", # Color for NA values - limits = c(min_count, max_count) # Define the range for the color scale + colours = c("#D73027", "#F0F0F0", "#4575B4"), + values = if ((abs(min_f) + max_f) > 0) c(0, abs(min_f)/(abs(min_f)+max_f), 1) else c(0, 0.5, 1), + na.value = "grey35", + limits = c(min_f, max_f) + ) + + scale_x_discrete( + labels = function(x) { + num <- suppressWarnings(as.numeric(gsub("[^0-9]", "", x))) + ifelse(num > max_orig_pos, " ", x) + }, + expand = expansion(mult = c(0, 0)) # no extra margin area ) + - scale_x_discrete(labels = function(x) { - numeric_pos <- as.numeric(gsub("[^0-9]", "", x)) - ifelse(numeric_pos > max_position, " ", x) - }) + geom_tile() + + # Solid white block covering the tail (no pattern / no seams) + { if (nrow(rect)) geom_rect(data = rect, inherit.aes = FALSE, + aes(xmin = xmin, xmax = xmax, ymin = ymin, ymax = ymax), + fill = "white", color = NA) } + + geom_segment( + data = syn, + aes(x = x - 0.485, xend = x + 0.485, y = y - 0.485, yend = y + 0.485), + linewidth = 0.2, inherit.aes = FALSE, color = "grey10" + ) + + theme_minimal() + + labs(title = title_text, x = "Wild-type amino acid", y = "Mutant amino acid", fill = "Fitness") + + theme( + plot.title = element_text(size = 16, face = "bold"), + axis.text.x = element_text(angle = 90, hjust = 1, vjust = 0.5, size = 10), + axis.text.y = element_text(size = 10), + axis.title.x = element_text(size = 14), + axis.title.y = element_text(size = 14), + legend.title = element_text(size = 12), + legend.text = element_text(size = 10), + panel.grid.major = element_blank(), + panel.grid.minor = element_blank(), + strip.text = element_blank(), + strip.background = element_blank(), + panel.spacing = grid::unit(0.2, "lines") + ) + + facet_wrap(~ row_group, scales = "free_x", ncol = 1) +} - # Add diagonal lines for synonymous mutations using geom_segment - geom_segment(data = syn_positions[syn_positions$position <= max_position, ], - aes(x = x - 0.485, xend = x + 0.485, - y = y - 0.485, yend = y + 0.485, color = synonymous), - size = 0.2) + +# ---------- main callable ---------- +# fitness_table_path : path to fitness_estimation.tsv +# wt_seq_aa_txt_path : path to TXT file containing WT AA sequence (one line) +# output_pdf_path : output PDF (default "fitness_heatmap.pdf") +# positions_per_row : default 75 +run_fitness_rescaled_heatmaps <- function(fitness_table_path, + wt_seq_aa_txt_path, + output_pdf_path = "fitness_heatmap.pdf", + positions_per_row = 75) { - # Manuelle Farbskala für die diagonalen Linien - scale_color_manual(values = c("TRUE" = "grey10"), labels = c("TRUE" = "")) + + df <- read.table( + fitness_table_path, sep = "\t", header = TRUE, + check.names = FALSE, quote = "", comment.char = "" + ) - theme_minimal() + - labs(title = "Heatmap of Fitness Estimations for Mutations", x = "Wild-type Amino Acid", y = "Mutant Amino Acid", fill = "Fitness", color = "Synonymous Mutation") + - theme(plot.title = element_text(size = 16, face = "bold"), - axis.text.x = element_text(angle = 90, hjust = 1, vjust = 0.5, size = 12), - axis.text.y = element_text(size = 12), # Larger y-axis labels - axis.title.x = element_text(size = 16), - axis.title.y = element_text(size = 16), - legend.title = element_text(size = 14), # Larger legend title - legend.text = element_text(size = 12), # Larger legend text - panel.spacing = unit(0.1, "lines"), # Adjust panel spacing - strip.text = element_blank(), # Remove row group labels (facet numbers) - strip.background = element_blank(), - panel.grid.major = element_blank(), # Remove major grid lines - panel.grid.minor = element_blank()) + # Remove minor grid lines - facet_wrap(~ row_group, scales = "free_x", ncol = 1) + # Group by 75 positions per row - theme(panel.spacing = unit(0.2, "lines")) - - heatmap_plot <- heatmap_plot + - geom_point(data = heatmap_data_long, aes(size = ""), colour = "black", alpha = 0) # Invisible points for legend - heatmap_plot <- heatmap_plot + - guides(size = guide_legend(paste("Dropout (Counts <", threshold, ")"), override.aes = list(shape = 15, size = 8, colour = "grey35", alpha = 1))) # Define Legend for Dropouts - - # Save the heatmap plot - if (img_format == "pdf") { - ggsave(output_pdf_path, plot = heatmap_plot, width = 16, height = plot_height, dpi = 150, device = cairo_pdf) - } else { - ggsave(output_pdf_path, plot = heatmap_plot, width = 16, height = plot_height, dpi = 150) + wt_aa_col <- find_col(df, c("wt aa", "wt_aa", "wt")) + pos_col <- find_col(df, c("pos", "position")) + mut_aa_col <- find_col(df, c("mut aa", "mut_aa", "aa")) + rescaled_cols <- get_rescaled_cols(df) + + wt_seq_aa <- read_wt_seq_aa_txt(wt_seq_aa_txt_path) + + plots <- list() + + ## 1) Mean first – use existing "mean fitness" column if available + mean_col <- find_mean_col(df) + if (is.null(mean_col)) { + df$`rescaled_fitness_mean` <- if (length(rescaled_cols) == 1) df[[rescaled_cols[1]]] else rowMeans(df[, rescaled_cols], na.rm = TRUE) + mean_col <- "rescaled_fitness_mean" } + long_df_mean <- build_heatmap_long(df, wt_aa_col, pos_col, mut_aa_col, mean_col, + positions_per_row, wt_seq_aa) + plots[[length(plots) + 1]] <- list( + title = sprintf("Fitness — mean of %d replicate(s)", length(rescaled_cols)), + data = long_df_mean + ) - if (file.exists(output_pdf_path)) { - print("Heatmap image successfully created!") - } else { - print("Error: Heatmap image was not created.") + ## 2) Then individual replicates + for (i in seq_along(rescaled_cols)) { + col <- rescaled_cols[i] + long_df <- build_heatmap_long(df, wt_aa_col, pos_col, mut_aa_col, col, + positions_per_row, wt_seq_aa) + plots[[length(plots) + 1]] <- list( + title = sprintf("Fitness — rep%d", i), + data = long_df + ) } -} -# Test the function -fitness_heatmap( - "/Users/benjaminwehnert/CRG/DMS_QC/howard/prepared_gatk_for_fitness.csv", - "/Users/benjaminwehnert/CRG/DMS_QC/howard/fitness_heatmap.pdf", - threshold = 3 -) - -#test on gid1a data -fitness_heatmap( - "/Users/benjaminwehnert/Downloads/dimsum_fitness_gid1a/prepared_gatk_for_fitness.csv", - "/Users/benjaminwehnert/Downloads/dimsum_fitness_gid1a/fitness_heatmap.pdf", - threshold = 3 -) + # Device height: (#row groups × 4) + page_heights <- vapply(plots, function(p) max(p$data$row_group, na.rm = TRUE), numeric(1)) + device_height <- max(4, as.numeric(page_heights) * 4, na.rm = TRUE) + + grDevices::pdf(output_pdf_path, width = 16, height = device_height) + on.exit(try(grDevices::dev.off(), silent = TRUE), add = TRUE) + for (p in plots) print(plot_heatmap(p$data, p$title, positions_per_row)) + invisible(TRUE) +} diff --git a/modules/local/fitness/fitness_standard.nf b/modules/local/fitness/fitness_standard.nf index c222833..4606077 100644 --- a/modules/local/fitness/fitness_standard.nf +++ b/modules/local/fitness/fitness_standard.nf @@ -86,3 +86,48 @@ process FITNESS_QC { EOF """ } + + + +process FITNESS_HEATMAP { + tag { sample.sample } + label 'process_single' + + conda "${moduleDir}/environment.yml" + container "${ workflow.containerEngine == 'singularity' + ? 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:ce2ba7ad7f6e7f2c' + : 'community.wave.seqera.io/library/bioconductor-biostrings_r-base_r-biocmanager_r-dplyr_pruned:0fd2e39a5bf2ecaa' }" + + input: + tuple val(sample), path(fitness_estimation_tsv) // from FITNESS_CALCULATION + tuple val(sample), path(wt_seq) // WT sequence + path script // fitness_plots.R + + output: + tuple val(sample), path("fitness_heatmap.pdf"), emit: fitness_heatmap + path "versions.yml", emit: versions + + script: + """ + set -euo pipefail + + R_version=\$(R --version | head -n 1 | sed 's/^R version //') + + Rscript -e "source('$script'); run_fitness_rescaled_heatmaps( + '$fitness_estimation_tsv', '$wt_seq')" + + cat > versions.yml < versions.yml <<'EOF' + FITNESS_PLOTS: + stub-version: "0.0.0" + EOF + """ +} diff --git a/workflows/deepmutscan.nf b/workflows/deepmutscan.nf index 1ae7d40..907e93d 100644 --- a/workflows/deepmutscan.nf +++ b/workflows/deepmutscan.nf @@ -25,6 +25,7 @@ include { EXPDESIGN_FITNESS } from '../modules/local/fitness/fitne include { FIND_SYNONYMOUS_MUTATION } from '../modules/local/fitness/find_synonymous_mutation' include { FITNESS_CALCULATION } from '../modules/local/fitness/fitness_standard' include { FITNESS_QC } from '../modules/local/fitness/fitness_standard' +include { FITNESS_HEATMAP } from '../modules/local/fitness/fitness_standard' include { RUN_DIMSUM } from '../modules/local/fitness/run_dimsum' include { paramsSummaryMap } from 'plugin/nf-schema' include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' @@ -122,6 +123,7 @@ R("modules/local/dmsanalysis/bin/merge_counts.R").set { merge_counts_script_ch } R("modules/local/dmsanalysis/bin/dimsum_experimentalDesign.R").set { exp_design_ch } R("modules/local/dmsanalysis/bin/fitness_calculation.R").set { fitness_calculation_script_ch } R("modules/local/dmsanalysis/bin/fitness_QC.R").set { fitness_QC_script_ch } 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z{XXh}-YVdyl!vc6j{_iBeR5XCBME+<^N4_-+BtH3BJQINT`Bc0uBv8v>XX=0?~_$! zOr2fNHQ`5`$L!BU#f-f!DQ;rNHnycY&rkGb+dg!}vidwm0+EqB+5 zh)mA-W5BpxPxH^$=krR7&rjv^>dfq1w4PfS9-6Xlt_~{e>Z|8{jf)*w9Dl35y^nGmHm|lNlTkQ9$=hEI~QK@$0NO!_~F=WQf8MFH# zSzK zam7t^2G@d$N8gW{U|cFDUSGpl7(VNL1CvTW?UA6^lvgRZ;=ZCjV~k6s3o)ozT66bN z{h;hfk0psdl2u(b7Qxjw;d_{OZ0b>{kk(vk(f3(3(rX1tl&j$;Rduo^%_GPk$ z%rPm~g>zS7;W)miT9}@^*yQ}6E$5~esb4Q<#g5+!ns9D;(SAO6)tW!*VwxBJYf#Co zh)TGjVoIALXFvVI$J-f1gHdQz(JAj8y%@BXMS9`~gRp~3m*761SM From 6c15247059c7d8c99cb6874c0df3731821957dd6 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 3 Nov 2025 16:34:44 +0100 Subject: [PATCH 281/317] Update counts_heatmap.R --- modules/local/dmsanalysis/bin/counts_heatmap.R | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/dmsanalysis/bin/counts_heatmap.R b/modules/local/dmsanalysis/bin/counts_heatmap.R index f685ef5..68a85a7 100644 --- a/modules/local/dmsanalysis/bin/counts_heatmap.R +++ b/modules/local/dmsanalysis/bin/counts_heatmap.R @@ -70,9 +70,9 @@ counts_heatmap <- function(input_csv_path, threshold = 3, output_pdf_path, img_f mutate(synonymous = mut_aa == wt_aa) # Definiere die korrekte Reihenfolge der Aminosäuren - amino_acid_order <- c("G", "A", "V", "L", "M", "I", "F", + amino_acid_order <- rev(c("G", "A", "V", "L", "M", "I", "F", "Y", "W", "K", "R", "H", "D", "E", - "S", "T", "C", "N", "Q", "P", "*") + "S", "T", "C", "N", "Q", "P", "*")) heatmap_data_long <- heatmap_data_long %>% mutate(mut_aa = factor(mut_aa, levels = amino_acid_order)) From ab17ed058757e4be04f6c30b5feec3282e8cdae1 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 3 Nov 2025 16:34:58 +0100 Subject: [PATCH 282/317] Update counts_per_cov_heatmap.R --- modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R b/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R index b51668e..2a98865 100644 --- a/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R +++ b/modules/local/dmsanalysis/bin/counts_per_cov_heatmap.R @@ -70,9 +70,9 @@ counts_per_cov_heatmap <- function(input_csv_path, threshold = 3, output_pdf_pat mutate(synonymous = mut_aa == wt_aa) # Definiere die korrekte Reihenfolge der Aminosäuren - amino_acid_order <- c("G", "A", "V", "L", "M", "I", "F", + amino_acid_order <- rev(c("G", "A", "V", "L", "M", "I", "F", "Y", "W", "K", "R", "H", "D", "E", - "S", "T", "C", "N", "Q", "P", "*") + "S", "T", "C", "N", "Q", "P", "*")) heatmap_data_long <- heatmap_data_long %>% mutate(mut_aa = factor(mut_aa, levels = amino_acid_order)) From ced474eb003ad80fdea066e4c897d748ff2b0889 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 3 Nov 2025 16:35:38 +0100 Subject: [PATCH 283/317] Update fitness_heatmap.R --- modules/local/dmsanalysis/bin/fitness_heatmap.R | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/modules/local/dmsanalysis/bin/fitness_heatmap.R b/modules/local/dmsanalysis/bin/fitness_heatmap.R index 348fa9f..28a405a 100644 --- a/modules/local/dmsanalysis/bin/fitness_heatmap.R +++ b/modules/local/dmsanalysis/bin/fitness_heatmap.R @@ -123,9 +123,9 @@ build_heatmap_long <- function(df, } syn_segments <- function(d, positions_per_row = 75) { - amino_order <- c("G", "A", "V", "L", "M", "I", "F", + amino_order <- rev(c("G", "A", "V", "L", "M", "I", "F", "Y", "W", "K", "R", "H", "D", "E", - "S", "T", "C", "N", "Q", "P", "*") + "S", "T", "C", "N", "Q", "P", "*")) d %>% mutate( mut_aa = factor(mut_aa, levels = amino_order), @@ -158,9 +158,9 @@ white_tail_rects <- function(d, positions_per_row = 75) { } plot_heatmap <- function(d, title_text, positions_per_row = 75) { - amino_order <- c("G", "A", "V", "L", "M", "I", "F", + amino_order <- rev(c("G", "A", "V", "L", "M", "I", "F", "Y", "W", "K", "R", "H", "D", "E", - "S", "T", "C", "N", "Q", "P", "*") + "S", "T", "C", "N", "Q", "P", "*")) d <- d %>% mutate(mut_aa = factor(mut_aa, levels = amino_order)) min_f <- suppressWarnings(min(d$fitness, na.rm = TRUE)); if (!is.finite(min_f)) min_f <- 0 From 059df716308a5f91675301168aa88fca285c182a Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Mon, 3 Nov 2025 22:10:59 +0100 Subject: [PATCH 284/317] new structure of output folders --- conf/modules.config | 10 +++++----- 1 file changed, 5 insertions(+), 5 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 5f19e0e..33b03f5 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -118,7 +118,7 @@ process { withName: 'GATK_GATKTOFITNESS' { publishDir = [ - path: "${params.outdir}/fitness/DiMSum/single_counts", + path: "${params.outdir}/fitness/DiMSum_results/single_rep_counts", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] @@ -150,7 +150,7 @@ process { withName: 'FITNESS_CALCULATION' { publishDir = [ - path: "${params.outdir}/fitness/output", + path: "${params.outdir}/fitness/default_results", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] @@ -158,7 +158,7 @@ process { withName: 'FITNESS_QC' { publishDir = [ - path: "${params.outdir}/fitness/output", + path: "${params.outdir}/fitness/default_results", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] @@ -166,7 +166,7 @@ process { withName: 'FITNESS_HEATMAP' { publishDir = [ - path: "${params.outdir}/fitness/output", + path: "${params.outdir}/fitness/default_results", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] @@ -174,7 +174,7 @@ process { withName: 'RUN_DIMSUM' { publishDir = [ - path: "${params.outdir}/fitness/DiMSum", + path: "${params.outdir}/fitness/DiMSum_results", mode: 'copy', saveAs: { filename -> filename == 'versions.yml' ? null : filename } ] From b89726a1f4abd84e430a579f5641857b2f6566ef Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 6 Nov 2025 12:41:10 +0100 Subject: [PATCH 285/317] Update README.md --- README.md | 33 +++++++++++++++++++-------------- 1 file changed, 19 insertions(+), 14 deletions(-) diff --git a/README.md b/README.md index acc6c8f..63e41eb 100644 --- a/README.md +++ b/README.md @@ -68,11 +68,12 @@ To execute **nf-core/deepmutscan**, run the basic command: nextflow run nf-core/deepmutscan \ -profile singularity,local \ --input ./input.csv \ - --outdir ./results \ - --fasta ./ref.fa \ --reading-frame 1-300 \ + --fasta ./ref.fa \ --mutagenesis NNK-NNS \ - --seq-rarefaction false + --run_seqdepth false \ + --fitness true \ + --outdir ./results ``` ### Required parameters @@ -84,15 +85,16 @@ nextflow run nf-core/deepmutscan \ | `--fasta` | Reference FASTA file | | `--reading_frame` | Start and end nucleotide (e.g. `1-300`) | -### Optional parameters +### Optional parameters *(in development)* | Parameter | Default | Description | |------------------------|-------------|-------------------------------------------------| -| `--read-align` | `bwa-mem` | Read aligner | -| `--mutagenesis` | `NNK-NNS` | Deep mutational scanning strategy used | -| `--seq-rarefaction` | `false` | Estimate sequencing saturation by rarefaction | -| `--error-estimation` | `input` | Error model used to correct 1nt counts | -| `--fitness-estimation` | `dimsum` | Downstream fitness inference module | +| `--run_seqdepth` | `false` | Estimate sequencing saturation by rarefaction | +| `--fitness` | `false` | Default fitness inference module | +| `--dimsum` | `false` | Optional fitness inference module *(AMD/x86_64 systems only)* | +| `--mutagenesis` | `NNK-NNS` | Deep mutational scanning strategy used *(in development)* | +| `--error-estimation` | `input` | Error model used to correct 1nt counts *(in development)* | +| `--read-align` | `bwa-mem` | Read aligner *(in development)* | More options and advanced configuration: [see vignette](link). For further information or help, don't hesitate to get in touch on the [Slack `#deepmutscan` channel](https://nfcore.slack.com/channels/deepmutscan) (you can join with [this invite](https://nf-co.re/join/slack)). @@ -117,11 +119,14 @@ After execution, the pipeline creates the following directory structure: ``` results/ -├── plots/ # PDF visualizations: coverage, variant heatmaps, etc. -├── intermediate_files/ # Raw alignments, filtered variant tables, QC reports -├── final_files/ # Fitness and error tables from downstream tools -├── timeline.html # Runtime timeline -└── report.html # Summary report incl. resource and CO₂ usage +├── fastqc/ # Individual HTML reports for specified fastq files, raw sequencing QC +├── fitness/ # Merged variant count tables, fitness and error estimates, replicate correlations and heatmaps +├── intermediate_files/ # Raw alignments, raw and pre-filtered variant count tables, QC reports +├── library_QC/ # Sample-specific PDF visualizations: position-wise sequencing coverage, count heatmaps, etc. +├── multiqc/ # Shared HTML reports for all fastq files, raw sequencing QC +├── pipelineinfo/ # Nextflow helper files for timeline and summary report generation +├── timeline.html # Nextflow timeline for all tasks +└── report.html # Nextflow summary report incl. detailed CPU and memory usage per for all tasks ``` --- From 636d84f4bb33cb14a691ad87a16f53f04648ce29 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 6 Nov 2025 12:47:27 +0100 Subject: [PATCH 286/317] Update pipeline_steps.md --- docs/pipeline_steps.md | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/docs/pipeline_steps.md b/docs/pipeline_steps.md index 83c18b5..670a7dc 100644 --- a/docs/pipeline_steps.md +++ b/docs/pipeline_steps.md @@ -49,11 +49,11 @@ Future versions may offer additional options depending on sequencing type and er ------------------------------------------------------------------------ -## 4. Mutation Counting +## 4. Variant Counting -Aligned, non-wildtype consensus reads are screened for exact, base-level mismatches. nf-core/deepmutscan uses a custom Python implementation to count the occurrences of all high-quality single, double, triple, and higher-order nucleotide changes between each read and the reference ORF. +Aligned, non-wildtype consensus reads are screened for exact, base-level mismatches. nf-core/deepmutscan currently uses the popular [**GATK AnalyzeSaturationMutagenesis**](https://gatk.broadinstitute.org/hc/en-us/articles/360037594771-AnalyzeSaturationMutagenesis-BETA) function to count occurrences of all single, double, triple, and higher-order nucleotide changes between each read and the reference ORF. -Unlike the popular [**GATK AnalyzeSaturationMutagenesis**](https://gatk.broadinstitute.org/hc/en-us/articles/360037594771-AnalyzeSaturationMutagenesis-BETA) function, nf-core/deepmutscan allows users to specify a minimum base quality cutoff for mutations to be included in the final count table (default: Q30). +We are currently working on the nf-core/deepmutscan implementation of a much lighter, alternative Python implementation for mutation counting. In this script, users will be allowed to specify a minimum base quality cutoff for mutations to be included in the final count table (default: Q30) – an option not available in GATK. ------------------------------------------------------------------------ From 3217b2fe1da95ea55026fcc2f3be07a3bfb0c15a Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 6 Nov 2025 12:48:50 +0100 Subject: [PATCH 287/317] Update samplesheet.csv --- assets/samplesheet.csv | 13 ++++--------- 1 file changed, 4 insertions(+), 9 deletions(-) diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv index 7477367..b176284 100644 --- a/assets/samplesheet.csv +++ b/assets/samplesheet.csv @@ -1,10 +1,5 @@ sample,type,replicate,file1,file2 -a_1,input,1,/path1/forward.fastq,/path1/reverse.fastq -a_1,input,2,/path2/forward.fastq,/path2/reverse.fastq -a_1,output,1,/path3/blabla_se.bam, -a_3,input,1,/path4/forward.fastq,/path4/reverse.fastq -a_3,input,2,/path5/forward.fastq,/path5/reverse.fastq -a_3,output,1,/path6/forward.fastq,/path6/reverse.fastq -a_3,output,2,/path7/forward.fastq,/path7/reverse.fastq -b_1,quality,1,/path8/single_end.fastq,/path8/reverse.fastq -b_1,quality,2,/path9/forward.fastq,/path9/reverse.fastq +a_1,input,1,/path1/forward1.fastq.gz,/path1/reverse1.fastq.gz +a_1,input,2,/path2/forward2.fastq.gz,/path2/reverse2.fastq.gz +a_3,output,1,/path4/forward3.fastq.gz,/path4/reverse3.fastq.gz +a_3,output,2,/path5/forward4.fastq.gz,/path5/reverse4.fastq.gz From dfb4e013e5538a3b18a81da194162e0a8532e6bd Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 6 Nov 2025 12:49:19 +0100 Subject: [PATCH 288/317] Update samplesheet.csv --- assets/samplesheet.csv | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv index b176284..a658721 100644 --- a/assets/samplesheet.csv +++ b/assets/samplesheet.csv @@ -1,5 +1,5 @@ sample,type,replicate,file1,file2 -a_1,input,1,/path1/forward1.fastq.gz,/path1/reverse1.fastq.gz -a_1,input,2,/path2/forward2.fastq.gz,/path2/reverse2.fastq.gz -a_3,output,1,/path4/forward3.fastq.gz,/path4/reverse3.fastq.gz -a_3,output,2,/path5/forward4.fastq.gz,/path5/reverse4.fastq.gz +a_1,input,1,/reads/forward1.fastq.gz,/reads/reverse1.fastq.gz +a_1,input,2,/reads/forward2.fastq.gz,/reads/reverse2.fastq.gz +a_1,output,1,/reads/forward3.fastq.gz,/reads/reverse3.fastq.gz +a_1,output,2,/reads/forward4.fastq.gz,/reads/reverse4.fastq.gz From eac51b0ae135831ce817162817d259d098f485d9 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 6 Nov 2025 12:49:36 +0100 Subject: [PATCH 289/317] Update samplesheet.csv --- assets/samplesheet.csv | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/assets/samplesheet.csv b/assets/samplesheet.csv index a658721..5b5d503 100644 --- a/assets/samplesheet.csv +++ b/assets/samplesheet.csv @@ -1,5 +1,5 @@ sample,type,replicate,file1,file2 -a_1,input,1,/reads/forward1.fastq.gz,/reads/reverse1.fastq.gz -a_1,input,2,/reads/forward2.fastq.gz,/reads/reverse2.fastq.gz -a_1,output,1,/reads/forward3.fastq.gz,/reads/reverse3.fastq.gz -a_1,output,2,/reads/forward4.fastq.gz,/reads/reverse4.fastq.gz +ORF1,input,1,/reads/forward1.fastq.gz,/reads/reverse1.fastq.gz +ORF1,input,2,/reads/forward2.fastq.gz,/reads/reverse2.fastq.gz +ORF1,output,1,/reads/forward3.fastq.gz,/reads/reverse3.fastq.gz +ORF1,output,2,/reads/forward4.fastq.gz,/reads/reverse4.fastq.gz From 61a60c5ba07504c7b6a815af6e40d8959f78a2f9 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Thu, 6 Nov 2025 19:59:56 +0100 Subject: [PATCH 290/317] Update README.md --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 63e41eb..cea205a 100644 --- a/README.md +++ b/README.md @@ -68,7 +68,7 @@ To execute **nf-core/deepmutscan**, run the basic command: nextflow run nf-core/deepmutscan \ -profile singularity,local \ --input ./input.csv \ - --reading-frame 1-300 \ + --reading_frame 1-300 \ --fasta ./ref.fa \ --mutagenesis NNK-NNS \ --run_seqdepth false \ From cf02d892f9c799c97705372407b6235b9f4fd283 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Fri, 7 Nov 2025 14:58:53 +0100 Subject: [PATCH 291/317] Update README.md --- README.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/README.md b/README.md index cea205a..a041439 100644 --- a/README.md +++ b/README.md @@ -70,7 +70,7 @@ nextflow run nf-core/deepmutscan \ --input ./input.csv \ --reading_frame 1-300 \ --fasta ./ref.fa \ - --mutagenesis NNK-NNS \ + --mutagenesis max_diff_to_wt \ --run_seqdepth false \ --fitness true \ --outdir ./results @@ -92,7 +92,7 @@ nextflow run nf-core/deepmutscan \ | `--run_seqdepth` | `false` | Estimate sequencing saturation by rarefaction | | `--fitness` | `false` | Default fitness inference module | | `--dimsum` | `false` | Optional fitness inference module *(AMD/x86_64 systems only)* | -| `--mutagenesis` | `NNK-NNS` | Deep mutational scanning strategy used *(in development)* | +| `--mutagenesis` | `max_diff_to_wt` | Deep mutational scanning strategy used *(in development)* | | `--error-estimation` | `input` | Error model used to correct 1nt counts *(in development)* | | `--read-align` | `bwa-mem` | Read aligner *(in development)* | From 9b7ec458c65249e3af9249a5d7deae6348c67893 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Fri, 7 Nov 2025 14:59:25 +0100 Subject: [PATCH 292/317] Update README.md --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index a041439..817996e 100644 --- a/README.md +++ b/README.md @@ -93,7 +93,7 @@ nextflow run nf-core/deepmutscan \ | `--fitness` | `false` | Default fitness inference module | | `--dimsum` | `false` | Optional fitness inference module *(AMD/x86_64 systems only)* | | `--mutagenesis` | `max_diff_to_wt` | Deep mutational scanning strategy used *(in development)* | -| `--error-estimation` | `input` | Error model used to correct 1nt counts *(in development)* | +| `--error-estimation` | `wt_sequencing` | Error model used to correct 1nt counts *(in development)* | | `--read-align` | `bwa-mem` | Read aligner *(in development)* | More options and advanced configuration: [see vignette](link). For further information or help, don't hesitate to get in touch on the [Slack `#deepmutscan` channel](https://nfcore.slack.com/channels/deepmutscan) (you can join with [this invite](https://nf-co.re/join/slack)). From 1ad0fd94caca150b952b853f5b009cb6767095f8 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Sun, 9 Nov 2025 15:40:33 +0100 Subject: [PATCH 293/317] automatic changes --- .github/CONTRIBUTING.md | 2 +- nextflow.config | 8 ++++++++ nextflow_schema.json | 3 +-- 3 files changed, 10 insertions(+), 3 deletions(-) diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index 00805c7..12dd4a0 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -10,7 +10,7 @@ However, don't be put off by this template - other more general issues and sugge Contributions to the code are even more welcome ;) > [!NOTE] -> If you need help using or modifying nf-core/deepmutscan then the best place to ask is on the nf-core Slack [#deepmutscan](https://nfcore.slack.com/channels/deepmutscan) channel ([join our Slack here](https://nf-co.re/join/slack)) +> If you need help using or modifying nf-core/deepmutscan then the best place to ask is on the nf-core Slack [#deepmutscan](https://nfcore.slack.com/channels/deepmutscan) channel ([join our Slack here](https://nf-co.re/join/slack)). ## Contribution workflow diff --git a/nextflow.config b/nextflow.config index 67771ec..c9a675f 100644 --- a/nextflow.config +++ b/nextflow.config @@ -25,6 +25,14 @@ params { max_multiqc_email_size = '25.MB' multiqc_methods_description = null + min_counts = 3 + mutagenesis_type = 'nnk' + fitness = false + dimsum = false + run_seqdepth = false + reading_frame = null + custom_codon_library = null + // Boilerplate options outdir = null publish_dir_mode = 'copy' diff --git a/nextflow_schema.json b/nextflow_schema.json index 9a52fb4..f67e925 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -44,8 +44,7 @@ "reading_frame": { "type": "string", "description": "Start and stop codon positions in the format 'start-stop', e.g., '352-1383'.", - "pattern": "^\\d+-\\d+$", - "default": "352-1383" + "pattern": "^\\d+-\\d+$" }, "min_counts": { "type": "integer", From e114b450e0bca07f75dc9980de5d65e604278095 Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Sun, 9 Nov 2025 16:16:03 +0100 Subject: [PATCH 294/317] further linting fixes --- nextflow.config | 2 +- nextflow_schema.json | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/nextflow.config b/nextflow.config index c9a675f..7984a5d 100644 --- a/nextflow.config +++ b/nextflow.config @@ -31,7 +31,7 @@ params { dimsum = false run_seqdepth = false reading_frame = null - custom_codon_library = null + custom_codon_library = '/NULL' // Boilerplate options outdir = null diff --git a/nextflow_schema.json b/nextflow_schema.json index f67e925..f1f0dd5 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -10,7 +10,7 @@ "type": "object", "fa_icon": "fas fa-terminal", "description": "Define where the pipeline should find input data and save output data.", - "required": ["input", "outdir"], + "required": ["input", "outdir", "reading_frame"], "properties": { "input": { "type": "string", From 32b3e9cf196cdc4aa95312f411aa511fcf93c4fe Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Sun, 9 Nov 2025 16:18:32 +0100 Subject: [PATCH 295/317] edited Required flags --- nextflow_schema.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index f1f0dd5..ccd9a85 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -10,7 +10,7 @@ "type": "object", "fa_icon": "fas fa-terminal", "description": "Define where the pipeline should find input data and save output data.", - "required": ["input", "outdir", "reading_frame"], + "required": ["input", "outdir", "fasta", "reading_frame"], "properties": { "input": { "type": "string", From f2c605f20e29a71daef375ee9023fb9fef8b9b2c Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Sun, 9 Nov 2025 16:23:42 +0100 Subject: [PATCH 296/317] linting in main.nf --- main.nf | 3 --- 1 file changed, 3 deletions(-) diff --git a/main.nf b/main.nf index bcee0db..5f339be 100644 --- a/main.nf +++ b/main.nf @@ -26,9 +26,6 @@ include { getGenomeAttribute } from './subworkflows/local/utils_nfcore_deep ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -// TODO nf-core: Remove this line if you don't need a FASTA file -// This is an example of how to use getGenomeAttribute() to fetch parameters -// from igenomes.config using `--genome` params.fasta = getGenomeAttribute('fasta') /* From c2c99d656f27204e016cfd40e13505119081871c Mon Sep 17 00:00:00 2001 From: BenjaminWehnert1008 Date: Sun, 9 Nov 2025 22:36:18 +0100 Subject: [PATCH 297/317] linting --- .../local/bamprocessing/bamfilteringdms.nf | 41 ------------------- modules/local/bamprocessing/premerge.nf | 16 -------- 2 files changed, 57 deletions(-) diff --git a/modules/local/bamprocessing/bamfilteringdms.nf b/modules/local/bamprocessing/bamfilteringdms.nf index 16603b7..6cef905 100644 --- a/modules/local/bamprocessing/bamfilteringdms.nf +++ b/modules/local/bamprocessing/bamfilteringdms.nf @@ -1,46 +1,18 @@ -// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) -// https://github.com/nf-core/modules/tree/master/modules/nf-core/ -// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: -// https://nf-co.re/join -// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. -// All other parameters MUST be provided using the "task.ext" directive, see here: -// https://www.nextflow.io/docs/latest/process.html#ext -// where "task.ext" is a string. -// Any parameters that need to be evaluated in the context of a particular sample -// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. -// TODO nf-core: Software that can be piped together SHOULD be added to separate module files -// unless there is a run-time, storage advantage in implementing in this way -// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: -// bwa mem | samtools view -B -T ref.fasta -// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty -// list (`[]`) instead of a file can be used to work around this issue. process BAMFILTER_DMS { tag "$meta.id" label 'process_single' - // TODO nf-core: List required Conda package(s). - // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). - // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. - // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. conda "${moduleDir}/environment.yml" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? 'https://depot.galaxyproject.org/singularity/samtools:1.21--h96c455f_1': 'biocontainers/samtools:1.21--h96c455f_1' }" input: - // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" - // MUST be provided as an input via a Groovy Map called "meta". - // This information may not be required in some instances e.g. indexing reference genome files: - // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf - // TODO nf-core: Where applicable please provide/convert compressed files as input/output - // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. tuple val(meta), path(bam) output: - // TODO nf-core: Named file extensions MUST be emitted for ALL output channels tuple val(meta), path("*.bam"), emit: bam - // TODO nf-core: List additional required output channels/values here path "versions.yml" , emit: versions when: @@ -49,15 +21,6 @@ process BAMFILTER_DMS { script: def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: Where possible, a command MUST be provided to obtain the version number of the software e.g. 1.10 - // If the software is unable to output a version number on the command-line then it can be manually specified - // e.g. https://github.com/nf-core/modules/blob/master/modules/nf-core/homer/annotatepeaks/main.nf - // Each software used MUST provide the software name and version number in the YAML version file (versions.yml) - // TODO nf-core: It MUST be possible to pass additional parameters to the tool as a command-line string via the "task.ext.args" directive - // TODO nf-core: If the tool supports multi-threading then you MUST provide the appropriate parameter - // using the Nextflow "task" variable e.g. "--threads $task.cpus" - // TODO nf-core: Please replace the example samtools command below with your module's command - // TODO nf-core: Please indent the command appropriately (4 spaces!!) to help with readability ;) """ samtools view -h -F 4 -F 256 -q 30 $bam | \ samtools view -h | \ @@ -75,10 +38,6 @@ process BAMFILTER_DMS { stub: def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" - // TODO nf-core: A stub section should mimic the execution of the original module as best as possible - // Have a look at the following examples: - // Simple example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bcftools/annotate/main.nf#L47-L63 - // Complex example: https://github.com/nf-core/modules/blob/818474a292b4860ae8ff88e149fbcda68814114d/modules/nf-core/bedtools/split/main.nf#L38-L54 """ touch ${prefix}.bam diff --git a/modules/local/bamprocessing/premerge.nf b/modules/local/bamprocessing/premerge.nf index 8f8b44a..af7e16f 100644 --- a/modules/local/bamprocessing/premerge.nf +++ b/modules/local/bamprocessing/premerge.nf @@ -1,19 +1,3 @@ -// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) -// https://github.com/nf-core/modules/tree/master/modules/nf-core/ -// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: -// https://nf-co.re/join -// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. -// All other parameters MUST be provided using the "task.ext" directive, see here: -// https://www.nextflow.io/docs/latest/process.html#ext -// where "task.ext" is a string. -// Any parameters that need to be evaluated in the context of a particular sample -// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. -// TODO nf-core: Software that can be piped together SHOULD be added to separate module files -// unless there is a run-time, storage advantage in implementing in this way -// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: -// bwa mem | samtools view -B -T ref.fasta -// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty -// list (`[]`) instead of a file can be used to work around this issue. process PREMERGE { tag "$meta.id" From 853ae9c2dfdc6ce1e597a3d3255136b3f4639a99 Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 10 Nov 2025 09:58:55 +0100 Subject: [PATCH 298/317] Create README.md --- testdata/README.md | 1 + 1 file changed, 1 insertion(+) create mode 100644 testdata/README.md diff --git a/testdata/README.md b/testdata/README.md new file mode 100644 index 0000000..219ecf1 --- /dev/null +++ b/testdata/README.md @@ -0,0 +1 @@ +4 x 50k sequencing reads (small pilot test data set) From 538c7584908089c7998193165ac2785fe21186ba Mon Sep 17 00:00:00 2001 From: Maximilian Stammnitz Date: Mon, 10 Nov 2025 09:59:33 +0100 Subject: [PATCH 299/317] Added test data directly to the repo --- testdata/input1_50k_1.fastq.gz | Bin 0 -> 1593369 bytes testdata/input1_50k_2.fastq.gz | Bin 0 -> 1666233 bytes testdata/input2_50k_1.fastq.gz | Bin 0 -> 1640235 bytes 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