From f1c1390029f2d6bc5d2253034f6498d159fca65f Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Fri, 30 Jun 2023 16:12:52 +0000 Subject: [PATCH 001/133] Template update for nf-core/tools version 2.9 --- .github/CONTRIBUTING.md | 1 - .github/ISSUE_TEMPLATE/bug_report.yml | 2 +- .github/workflows/awsfulltest.yml | 11 +- .github/workflows/awstest.yml | 10 +- .github/workflows/ci.yml | 2 +- .gitpod.yml | 5 + CHANGELOG.md | 2 +- CITATIONS.md | 6 + README.md | 6 +- assets/methods_description_template.yml | 12 +- assets/multiqc_config.yml | 4 +- assets/nf-core-crisprseq_logo_light.png | Bin 11593 -> 74216 bytes assets/slackreport.json | 2 +- conf/test_full.config | 2 - docs/usage.md | 6 +- lib/NfcoreSchema.groovy | 530 ------------------------ lib/NfcoreTemplate.groovy | 2 +- lib/WorkflowCrisprseq.groovy | 45 +- lib/WorkflowMain.groovy | 37 -- main.nf | 16 + nextflow.config | 54 ++- nextflow_schema.json | 36 +- workflows/crisprseq.nf | 25 +- 23 files changed, 177 insertions(+), 639 deletions(-) delete mode 100755 lib/NfcoreSchema.groovy diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index 3893df65..607441a1 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -116,4 +116,3 @@ To get started: Devcontainer specs: - [DevContainer config](.devcontainer/devcontainer.json) -- [Dockerfile](.devcontainer/Dockerfile) diff --git a/.github/ISSUE_TEMPLATE/bug_report.yml b/.github/ISSUE_TEMPLATE/bug_report.yml index a6a2106b..5be250d7 100644 --- a/.github/ISSUE_TEMPLATE/bug_report.yml +++ b/.github/ISSUE_TEMPLATE/bug_report.yml @@ -42,7 +42,7 @@ body: attributes: label: System information description: | - * Nextflow version _(eg. 22.10.1)_ + * Nextflow version _(eg. 23.04.0)_ * Hardware _(eg. HPC, Desktop, Cloud)_ * Executor _(eg. slurm, local, awsbatch)_ * Container engine: _(e.g. Docker, Singularity, Conda, Podman, Shifter, Charliecloud, or Apptainer)_ diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml index 0cae140d..92ebecb0 100644 --- a/.github/workflows/awsfulltest.yml +++ b/.github/workflows/awsfulltest.yml @@ -14,7 +14,7 @@ jobs: runs-on: ubuntu-latest steps: - name: Launch workflow via tower - uses: seqeralabs/action-tower-launch@v1 + uses: seqeralabs/action-tower-launch@v2 # TODO nf-core: You can customise AWS full pipeline tests as required # Add full size test data (but still relatively small datasets for few samples) # on the `test_full.config` test runs with only one set of parameters @@ -22,13 +22,18 @@ jobs: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} + revision: ${{ github.sha }} workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/crisprseq/work-${{ github.sha }} parameters: | { + "hook_url": "${{ secrets.MEGATESTS_ALERTS_SLACK_HOOK_URL }}", "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/crisprseq/results-${{ github.sha }}" } - profiles: test_full,aws_tower + profiles: test_full + - uses: actions/upload-artifact@v3 with: name: Tower debug log file - path: tower_action_*.log + path: | + tower_action_*.log + tower_action_*.json diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 6d64d2ba..0d650d8e 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -12,18 +12,22 @@ jobs: steps: # Launch workflow using Tower CLI tool action - name: Launch workflow via tower - uses: seqeralabs/action-tower-launch@v1 + uses: seqeralabs/action-tower-launch@v2 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} + revision: ${{ github.sha }} workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/crisprseq/work-${{ github.sha }} parameters: | { "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/crisprseq/results-test-${{ github.sha }}" } - profiles: test,aws_tower + profiles: test + - uses: actions/upload-artifact@v3 with: name: Tower debug log file - path: tower_action_*.log + path: | + tower_action_*.log + tower_action_*.json diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index c6c9113a..277385ff 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -24,7 +24,7 @@ jobs: strategy: matrix: NXF_VER: - - "22.10.1" + - "23.04.0" - "latest-everything" steps: - name: Check out pipeline code diff --git a/.gitpod.yml b/.gitpod.yml index 85d95ecc..25488dcc 100644 --- a/.gitpod.yml +++ b/.gitpod.yml @@ -1,4 +1,9 @@ image: nfcore/gitpod:latest +tasks: + - name: Update Nextflow and setup pre-commit + command: | + pre-commit install --install-hooks + nextflow self-update vscode: extensions: # based on nf-core.nf-core-extensionpack diff --git a/CHANGELOG.md b/CHANGELOG.md index 1d00cceb..6f507b61 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ 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 - [date] +## v2.0.0 - [date] Initial release of nf-core/crisprseq, created with the [nf-core](https://nf-co.re/) template. diff --git a/CITATIONS.md b/CITATIONS.md index 7093ef8f..287bcf20 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -12,7 +12,10 @@ - [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) + > Andrews, S. (2010). FastQC: A Quality Control Tool for High Throughput Sequence Data [Online]. Available online https://www.bioinformatics.babraham.ac.uk/projects/fastqc/. + - [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/) + > Ewels P, Magnusson M, Lundin S, Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016 Oct 1;32(19):3047-8. doi: 10.1093/bioinformatics/btw354. Epub 2016 Jun 16. PubMed PMID: 27312411; PubMed Central PMCID: PMC5039924. ## Software packaging/containerisation tools @@ -31,5 +34,8 @@ - [Docker](https://dl.acm.org/doi/10.5555/2600239.2600241) + > Merkel, D. (2014). Docker: lightweight linux containers for consistent development and deployment. Linux Journal, 2014(239), 2. doi: 10.5555/2600239.2600241. + - [Singularity](https://pubmed.ncbi.nlm.nih.gov/28494014/) + > Kurtzer GM, Sochat V, Bauer MW. Singularity: Scientific containers for mobility of compute. PLoS One. 2017 May 11;12(5):e0177459. doi: 10.1371/journal.pone.0177459. eCollection 2017. PubMed PMID: 28494014; PubMed Central PMCID: PMC5426675. diff --git a/README.md b/README.md index 9f069719..23db4ee3 100644 --- a/README.md +++ b/README.md @@ -2,7 +2,7 @@ [![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) -[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A522.10.1-23aa62.svg)](https://www.nextflow.io/) +[![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A523.04.0-23aa62.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/) @@ -66,11 +66,11 @@ nextflow run nf-core/crisprseq \ > provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; > see [docs](https://nf-co.re/usage/configuration#custom-configuration-files). -For more details, please refer to the [usage documentation](https://nf-co.re/crisprseq/usage) and the [parameter documentation](https://nf-co.re/crisprseq/parameters). +For more details and further functionality, please refer to the [usage documentation](https://nf-co.re/crisprseq/usage) and the [parameter documentation](https://nf-co.re/crisprseq/parameters). ## Pipeline output -To see the the results of a test run with a full size dataset refer to the [results](https://nf-co.re/crisprseq/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/crisprseq/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/crisprseq/output). diff --git a/assets/methods_description_template.yml b/assets/methods_description_template.yml index c10071b3..e2693af3 100644 --- a/assets/methods_description_template.yml +++ b/assets/methods_description_template.yml @@ -3,17 +3,21 @@ description: "Suggested text and references to use when describing pipeline usag section_name: "nf-core/crisprseq Methods Description" section_href: "https://github.com/nf-core/crisprseq" plot_type: "html" -## TODO nf-core: Update the HTML below to your prefered methods description, e.g. add publication citation for this pipeline +## 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/crisprseq v${workflow.manifest.version} ${doi_text} of the nf-core collection of workflows (Ewels et al., 2020).

+

Data was processed using nf-core/crisprseq 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}

References

Notes:
diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 4a692180..5f5e0401 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -1,7 +1,7 @@ report_comment: > - This report has been generated by the nf-core/crisprseq + This report has been generated by the nf-core/crisprseq analysis pipeline. 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100644 --- a/conf/test_full.config +++ b/conf/test_full.config @@ -10,8 +10,6 @@ ---------------------------------------------------------------------------------------- */ -cleanup = true - params { config_profile_name = 'Full test profile' config_profile_description = 'Full test dataset to check pipeline function' diff --git a/docs/usage.md b/docs/usage.md index cf8951d1..1a8c67bf 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -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/crisprseq --input samplesheet.csv --outdir --genome GRCh37 -profile docker +nextflow run nf-core/crisprseq --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. @@ -76,7 +76,8 @@ If you wish to repeatedly use the same parameters for multiple runs, rather than Pipeline settings can be provided in a `yaml` or `json` file via `-params-file `. > ⚠️ Do not use `-c ` to specify parameters as this will result in errors. Custom config files specified with `-c` must only be used for [tuning process resource specifications](https://nf-co.re/docs/usage/configuration#tuning-workflow-resources), other infrastructural tweaks (such as output directories), or module arguments (args). -> The above pipeline run specified with a params file in yaml format: + +The above pipeline run specified with a params file in yaml format: ```bash nextflow run nf-core/crisprseq -profile docker -params-file params.yaml @@ -88,7 +89,6 @@ with `params.yaml` containing: input: './samplesheet.csv' outdir: './results/' genome: 'GRCh37' -input: 'data' <...> ``` diff --git a/lib/NfcoreSchema.groovy b/lib/NfcoreSchema.groovy deleted file mode 100755 index 9b34804d..00000000 --- a/lib/NfcoreSchema.groovy +++ /dev/null @@ -1,530 +0,0 @@ -// -// This file holds several functions used to perform JSON parameter validation, help and summary rendering for the nf-core pipeline template. -// - -import nextflow.Nextflow -import org.everit.json.schema.Schema -import org.everit.json.schema.loader.SchemaLoader -import org.everit.json.schema.ValidationException -import org.json.JSONObject -import org.json.JSONTokener -import org.json.JSONArray -import groovy.json.JsonSlurper -import groovy.json.JsonBuilder - -class NfcoreSchema { - - // - // Resolve Schema path relative to main workflow directory - // - public static String getSchemaPath(workflow, schema_filename='nextflow_schema.json') { - return "${workflow.projectDir}/${schema_filename}" - } - - // - // Function to loop over all parameters defined in schema and check - // whether the given parameters adhere to the specifications - // - /* groovylint-disable-next-line UnusedPrivateMethodParameter */ - public static void validateParameters(workflow, params, log, schema_filename='nextflow_schema.json') { - def has_error = false - //~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~// - // Check for nextflow core params and unexpected params - def json = new File(getSchemaPath(workflow, schema_filename=schema_filename)).text - def Map schemaParams = (Map) new JsonSlurper().parseText(json).get('definitions') - def nf_params = [ - // Options for base `nextflow` command - 'bg', - 'c', - 'C', - 'config', - 'd', - 'D', - 'dockerize', - 'h', - 'log', - 'q', - 'quiet', - 'syslog', - 'v', - - // Options for `nextflow run` command - 'ansi', - 'ansi-log', - 'bg', - 'bucket-dir', - 'c', - 'cache', - 'config', - 'dsl2', - 'dump-channels', - 'dump-hashes', - 'E', - 'entry', - 'latest', - 'lib', - 'main-script', - 'N', - 'name', - 'offline', - 'params-file', - 'pi', - 'plugins', - 'poll-interval', - 'pool-size', - 'profile', - 'ps', - 'qs', - 'queue-size', - 'r', - 'resume', - 'revision', - 'stdin', - 'stub', - 'stub-run', - 'test', - 'w', - 'with-apptainer', - 'with-charliecloud', - 'with-conda', - 'with-dag', - 'with-docker', - 'with-mpi', - 'with-notification', - 'with-podman', - 'with-report', - 'with-singularity', - 'with-timeline', - 'with-tower', - 'with-trace', - 'with-weblog', - 'without-docker', - 'without-podman', - 'work-dir' - ] - def unexpectedParams = [] - - // Collect expected parameters from the schema - def expectedParams = [] - def enums = [:] - for (group in schemaParams) { - for (p in group.value['properties']) { - expectedParams.push(p.key) - if (group.value['properties'][p.key].containsKey('enum')) { - enums[p.key] = group.value['properties'][p.key]['enum'] - } - } - } - - for (specifiedParam in params.keySet()) { - // nextflow params - if (nf_params.contains(specifiedParam)) { - log.error "ERROR: You used a core Nextflow option with two hyphens: '--${specifiedParam}'. Please resubmit with '-${specifiedParam}'" - has_error = true - } - // unexpected params - def params_ignore = params.schema_ignore_params.split(',') + 'schema_ignore_params' - def expectedParamsLowerCase = expectedParams.collect{ it.replace("-", "").toLowerCase() } - def specifiedParamLowerCase = specifiedParam.replace("-", "").toLowerCase() - def isCamelCaseBug = (specifiedParam.contains("-") && !expectedParams.contains(specifiedParam) && expectedParamsLowerCase.contains(specifiedParamLowerCase)) - if (!expectedParams.contains(specifiedParam) && !params_ignore.contains(specifiedParam) && !isCamelCaseBug) { - // Temporarily remove camelCase/camel-case params #1035 - def unexpectedParamsLowerCase = unexpectedParams.collect{ it.replace("-", "").toLowerCase()} - if (!unexpectedParamsLowerCase.contains(specifiedParamLowerCase)){ - unexpectedParams.push(specifiedParam) - } - } - } - - //~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~// - // Validate parameters against the schema - InputStream input_stream = new File(getSchemaPath(workflow, schema_filename=schema_filename)).newInputStream() - JSONObject raw_schema = new JSONObject(new JSONTokener(input_stream)) - - // Remove anything that's in params.schema_ignore_params - raw_schema = removeIgnoredParams(raw_schema, params) - - Schema schema = SchemaLoader.load(raw_schema) - - // Clean the parameters - def cleanedParams = cleanParameters(params) - - // Convert to JSONObject - def jsonParams = new JsonBuilder(cleanedParams) - JSONObject params_json = new JSONObject(jsonParams.toString()) - - // Validate - try { - schema.validate(params_json) - } catch (ValidationException e) { - println '' - log.error 'ERROR: Validation of pipeline parameters failed!' - JSONObject exceptionJSON = e.toJSON() - printExceptions(exceptionJSON, params_json, log, enums) - println '' - has_error = true - } - - // Check for unexpected parameters - if (unexpectedParams.size() > 0) { - Map colors = NfcoreTemplate.logColours(params.monochrome_logs) - println '' - def warn_msg = 'Found unexpected parameters:' - for (unexpectedParam in unexpectedParams) { - warn_msg = warn_msg + "\n* --${unexpectedParam}: ${params[unexpectedParam].toString()}" - } - log.warn warn_msg - log.info "- ${colors.dim}Ignore this warning: params.schema_ignore_params = \"${unexpectedParams.join(',')}\" ${colors.reset}" - println '' - } - - if (has_error) { - Nextflow.error('Exiting!') - } - } - - // - // Beautify parameters for --help - // - public static String paramsHelp(workflow, params, command, schema_filename='nextflow_schema.json') { - Map colors = NfcoreTemplate.logColours(params.monochrome_logs) - Integer num_hidden = 0 - String output = '' - output += 'Typical pipeline command:\n\n' - output += " ${colors.cyan}${command}${colors.reset}\n\n" - Map params_map = paramsLoad(getSchemaPath(workflow, schema_filename=schema_filename)) - Integer max_chars = paramsMaxChars(params_map) + 1 - Integer desc_indent = max_chars + 14 - Integer dec_linewidth = 160 - desc_indent - for (group in params_map.keySet()) { - Integer num_params = 0 - String group_output = colors.underlined + colors.bold + group + colors.reset + '\n' - def group_params = params_map.get(group) // This gets the parameters of that particular group - for (param in group_params.keySet()) { - if (group_params.get(param).hidden && !params.show_hidden_params) { - num_hidden += 1 - continue; - } - def type = '[' + group_params.get(param).type + ']' - def description = group_params.get(param).description - def defaultValue = group_params.get(param).default != null ? " [default: " + group_params.get(param).default.toString() + "]" : '' - def description_default = description + colors.dim + defaultValue + colors.reset - // Wrap long description texts - // Loosely based on https://dzone.com/articles/groovy-plain-text-word-wrap - if (description_default.length() > dec_linewidth){ - List olines = [] - String oline = "" // " " * indent - description_default.split(" ").each() { wrd -> - if ((oline.size() + wrd.size()) <= dec_linewidth) { - oline += wrd + " " - } else { - olines += oline - oline = wrd + " " - } - } - olines += oline - description_default = olines.join("\n" + " " * desc_indent) - } - group_output += " --" + param.padRight(max_chars) + colors.dim + type.padRight(10) + colors.reset + description_default + '\n' - num_params += 1 - } - group_output += '\n' - if (num_params > 0){ - output += group_output - } - } - if (num_hidden > 0){ - output += colors.dim + "!! Hiding $num_hidden params, use --show_hidden_params to show them !!\n" + colors.reset - } - output += NfcoreTemplate.dashedLine(params.monochrome_logs) - return output - } - - // - // Groovy Map summarising parameters/workflow options used by the pipeline - // - public static LinkedHashMap paramsSummaryMap(workflow, params, schema_filename='nextflow_schema.json') { - // Get a selection of core Nextflow workflow options - def Map workflow_summary = [:] - if (workflow.revision) { - workflow_summary['revision'] = workflow.revision - } - workflow_summary['runName'] = workflow.runName - if (workflow.containerEngine) { - workflow_summary['containerEngine'] = workflow.containerEngine - } - if (workflow.container) { - workflow_summary['container'] = workflow.container - } - workflow_summary['launchDir'] = workflow.launchDir - workflow_summary['workDir'] = workflow.workDir - workflow_summary['projectDir'] = workflow.projectDir - workflow_summary['userName'] = workflow.userName - workflow_summary['profile'] = workflow.profile - workflow_summary['configFiles'] = workflow.configFiles.join(', ') - - // Get pipeline parameters defined in JSON Schema - def Map params_summary = [:] - def params_map = paramsLoad(getSchemaPath(workflow, schema_filename=schema_filename)) - for (group in params_map.keySet()) { - def sub_params = new LinkedHashMap() - def group_params = params_map.get(group) // This gets the parameters of that particular group - for (param in group_params.keySet()) { - if (params.containsKey(param)) { - def params_value = params.get(param) - def schema_value = group_params.get(param).default - def param_type = group_params.get(param).type - if (schema_value != null) { - if (param_type == 'string') { - if (schema_value.contains('$projectDir') || schema_value.contains('${projectDir}')) { - def sub_string = schema_value.replace('\$projectDir', '') - sub_string = sub_string.replace('\${projectDir}', '') - if (params_value.contains(sub_string)) { - schema_value = params_value - } - } - if (schema_value.contains('$params.outdir') || schema_value.contains('${params.outdir}')) { - def sub_string = schema_value.replace('\$params.outdir', '') - sub_string = sub_string.replace('\${params.outdir}', '') - if ("${params.outdir}${sub_string}" == params_value) { - schema_value = params_value - } - } - } - } - - // We have a default in the schema, and this isn't it - if (schema_value != null && params_value != schema_value) { - sub_params.put(param, params_value) - } - // No default in the schema, and this isn't empty - else if (schema_value == null && params_value != "" && params_value != null && params_value != false) { - sub_params.put(param, params_value) - } - } - } - params_summary.put(group, sub_params) - } - return [ 'Core Nextflow options' : workflow_summary ] << params_summary - } - - // - // Beautify parameters for summary and return as string - // - public static String paramsSummaryLog(workflow, params) { - Map colors = NfcoreTemplate.logColours(params.monochrome_logs) - String output = '' - def params_map = paramsSummaryMap(workflow, params) - def max_chars = paramsMaxChars(params_map) - for (group in params_map.keySet()) { - def group_params = params_map.get(group) // This gets the parameters of that particular group - if (group_params) { - output += colors.bold + group + colors.reset + '\n' - for (param in group_params.keySet()) { - output += " " + colors.blue + param.padRight(max_chars) + ": " + colors.green + group_params.get(param) + colors.reset + '\n' - } - output += '\n' - } - } - output += "!! Only displaying parameters that differ from the pipeline defaults !!\n" - output += NfcoreTemplate.dashedLine(params.monochrome_logs) - return output - } - - // - // Loop over nested exceptions and print the causingException - // - private static void printExceptions(ex_json, params_json, log, enums, limit=5) { - def causingExceptions = ex_json['causingExceptions'] - if (causingExceptions.length() == 0) { - def m = ex_json['message'] =~ /required key \[([^\]]+)\] not found/ - // Missing required param - if (m.matches()) { - log.error "* Missing required parameter: --${m[0][1]}" - } - // Other base-level error - else if (ex_json['pointerToViolation'] == '#') { - log.error "* ${ex_json['message']}" - } - // Error with specific param - else { - def param = ex_json['pointerToViolation'] - ~/^#\// - def param_val = params_json[param].toString() - if (enums.containsKey(param)) { - def error_msg = "* --${param}: '${param_val}' is not a valid choice (Available choices" - if (enums[param].size() > limit) { - log.error "${error_msg} (${limit} of ${enums[param].size()}): ${enums[param][0..limit-1].join(', ')}, ... )" - } else { - log.error "${error_msg}: ${enums[param].join(', ')})" - } - } else { - log.error "* --${param}: ${ex_json['message']} (${param_val})" - } - } - } - for (ex in causingExceptions) { - printExceptions(ex, params_json, log, enums) - } - } - - // - // Remove an element from a JSONArray - // - private static JSONArray removeElement(json_array, element) { - def list = [] - int len = json_array.length() - for (int i=0;i - if(raw_schema.keySet().contains('definitions')){ - raw_schema.definitions.each { definition -> - for (key in definition.keySet()){ - if (definition[key].get("properties").keySet().contains(ignore_param)){ - // Remove the param to ignore - definition[key].get("properties").remove(ignore_param) - // If the param was required, change this - if (definition[key].has("required")) { - def cleaned_required = removeElement(definition[key].required, ignore_param) - definition[key].put("required", cleaned_required) - } - } - } - } - } - if(raw_schema.keySet().contains('properties') && raw_schema.get('properties').keySet().contains(ignore_param)) { - raw_schema.get("properties").remove(ignore_param) - } - if(raw_schema.keySet().contains('required') && raw_schema.required.contains(ignore_param)) { - def cleaned_required = removeElement(raw_schema.required, ignore_param) - raw_schema.put("required", cleaned_required) - } - } - return raw_schema - } - - // - // Clean and check parameters relative to Nextflow native classes - // - private static Map cleanParameters(params) { - def new_params = params.getClass().newInstance(params) - for (p in params) { - // remove anything evaluating to false - if (!p['value']) { - new_params.remove(p.key) - } - // Cast MemoryUnit to String - if (p['value'].getClass() == nextflow.util.MemoryUnit) { - new_params.replace(p.key, p['value'].toString()) - } - // Cast Duration to String - if (p['value'].getClass() == nextflow.util.Duration) { - new_params.replace(p.key, p['value'].toString().replaceFirst(/d(?!\S)/, "day")) - } - // Cast LinkedHashMap to String - if (p['value'].getClass() == LinkedHashMap) { - new_params.replace(p.key, p['value'].toString()) - } - } - return new_params - } - - // - // This function tries to read a JSON params file - // - private static LinkedHashMap paramsLoad(String json_schema) { - def params_map = new LinkedHashMap() - try { - params_map = paramsRead(json_schema) - } catch (Exception e) { - println "Could not read parameters settings from JSON. $e" - params_map = new LinkedHashMap() - } - return params_map - } - - // - // Method to actually read in JSON file using Groovy. - // Group (as Key), values are all parameters - // - Parameter1 as Key, Description as Value - // - Parameter2 as Key, Description as Value - // .... - // Group - // - - private static LinkedHashMap paramsRead(String json_schema) throws Exception { - def json = new File(json_schema).text - def Map schema_definitions = (Map) new JsonSlurper().parseText(json).get('definitions') - def Map schema_properties = (Map) new JsonSlurper().parseText(json).get('properties') - /* Tree looks like this in nf-core schema - * definitions <- this is what the first get('definitions') gets us - group 1 - title - description - properties - parameter 1 - type - description - parameter 2 - type - description - group 2 - title - description - properties - parameter 1 - type - description - * properties <- parameters can also be ungrouped, outside of definitions - parameter 1 - type - description - */ - - // Grouped params - def params_map = new LinkedHashMap() - schema_definitions.each { key, val -> - def Map group = schema_definitions."$key".properties // Gets the property object of the group - def title = schema_definitions."$key".title - def sub_params = new LinkedHashMap() - group.each { innerkey, value -> - sub_params.put(innerkey, value) - } - params_map.put(title, sub_params) - } - - // Ungrouped params - def ungrouped_params = new LinkedHashMap() - schema_properties.each { innerkey, value -> - ungrouped_params.put(innerkey, value) - } - params_map.put("Other parameters", ungrouped_params) - - return params_map - } - - // - // Get maximum number of characters across all parameter names - // - private static Integer paramsMaxChars(params_map) { - Integer max_chars = 0 - for (group in params_map.keySet()) { - def group_params = params_map.get(group) // This gets the parameters of that particular group - for (param in group_params.keySet()) { - if (param.size() > max_chars) { - max_chars = param.size() - } - } - } - return max_chars - } -} diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy index 25a0a74a..408951ae 100755 --- a/lib/NfcoreTemplate.groovy +++ b/lib/NfcoreTemplate.groovy @@ -128,7 +128,7 @@ class NfcoreTemplate { def email_html = html_template.toString() // Render the sendmail template - def max_multiqc_email_size = params.max_multiqc_email_size as nextflow.util.MemoryUnit + def max_multiqc_email_size = (params.containsKey('max_multiqc_email_size') ? params.max_multiqc_email_size : 0) as nextflow.util.MemoryUnit def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, projectDir: "$projectDir", mqcFile: mqc_report, mqcMaxSize: max_multiqc_email_size.toBytes() ] def sf = new File("$projectDir/assets/sendmail_template.txt") def sendmail_template = engine.createTemplate(sf).make(smail_fields) diff --git a/lib/WorkflowCrisprseq.groovy b/lib/WorkflowCrisprseq.groovy index 243f5da0..dd252d59 100755 --- a/lib/WorkflowCrisprseq.groovy +++ b/lib/WorkflowCrisprseq.groovy @@ -11,6 +11,7 @@ class WorkflowCrisprseq { // Check and validate parameters // public static void initialise(params, log) { + genomeExistsError(params, log) @@ -46,15 +47,57 @@ class WorkflowCrisprseq { return yaml_file_text } - public static String methodsDescriptionText(run_workflow, mqc_methods_yaml) { + // + // Generate methods description for MultiQC + // + + public static String toolCitationText(params) { + + // TODO Optionally add in-text citation tools to this list. + // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "Tool (Foo et al. 2023)" : "", + // Uncomment function in methodsDescriptionText to render in MultiQC report + def citation_text = [ + "Tools used in the workflow included:", + "FastQC (Andrews 2010),", + "MultiQC (Ewels et al. 2016)", + "." + ].join(' ').trim() + + return citation_text + } + + public static String toolBibliographyText(params) { + + // TODO Optionally add bibliographic entries to this list. + // Can use ternary operators to dynamically construct based conditions, e.g. params["run_xyz"] ? "

  • Author (2023) Pub name, Journal, DOI
    • " : "", + // Uncomment function in methodsDescriptionText to render in MultiQC report + def reference_text = [ + "
  • Andrews S, (2010) FastQC, URL: https://www.bioinformatics.babraham.ac.uk/projects/fastqc/).
    • ", + "
  • Ewels, P., Magnusson, M., Lundin, S., & Käller, M. (2016). MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics , 32(19), 3047–3048. doi: /10.1093/bioinformatics/btw354
    • " + ].join(' ').trim() + + return reference_text + } + + public static String methodsDescriptionText(run_workflow, mqc_methods_yaml, params) { // Convert to a named map so can be used as with familar NXF ${workflow} variable syntax in the MultiQC YML file def meta = [:] meta.workflow = run_workflow.toMap() meta["manifest_map"] = run_workflow.manifest.toMap() + // Pipeline DOI meta["doi_text"] = meta.manifest_map.doi ? "(doi:     ${meta.manifest_map.doi})" : "" meta["nodoi_text"] = meta.manifest_map.doi ? "": "
  • If available, make sure to update the text to include the Zenodo DOI of version of the pipeline used.
    • " + // Tool references + meta["tool_citations"] = "" + meta["tool_bibliography"] = "" + + // TODO Only uncomment below if logic in toolCitationText/toolBibliographyText has been filled! + //meta["tool_citations"] = toolCitationText(params).replaceAll(", \\.", ".").replaceAll("\\. \\.", ".").replaceAll(", \\.", ".") + //meta["tool_bibliography"] = toolBibliographyText(params) + + def methods_text = mqc_methods_yaml.text def engine = new SimpleTemplateEngine() diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy index 72fbde90..a19a8241 100755 --- a/lib/WorkflowMain.groovy +++ b/lib/WorkflowMain.groovy @@ -20,40 +20,11 @@ class WorkflowMain { " https://github.com/${workflow.manifest.name}/blob/master/CITATIONS.md" } - // - // Generate help string - // - public static String help(workflow, params) { - def command = "nextflow run ${workflow.manifest.name} --input samplesheet.csv --genome GRCh37 -profile docker" - def help_string = '' - help_string += NfcoreTemplate.logo(workflow, params.monochrome_logs) - help_string += NfcoreSchema.paramsHelp(workflow, params, command) - help_string += '\n' + citation(workflow) + '\n' - help_string += NfcoreTemplate.dashedLine(params.monochrome_logs) - return help_string - } - - // - // Generate parameter summary log string - // - public static String paramsSummaryLog(workflow, params) { - def summary_log = '' - summary_log += NfcoreTemplate.logo(workflow, params.monochrome_logs) - summary_log += NfcoreSchema.paramsSummaryLog(workflow, params) - summary_log += '\n' + citation(workflow) + '\n' - summary_log += NfcoreTemplate.dashedLine(params.monochrome_logs) - return summary_log - } // // Validate parameters and print summary to screen // public static void initialise(workflow, params, log) { - // Print help to screen if required - if (params.help) { - log.info help(workflow, params) - System.exit(0) - } // Print workflow version and exit on --version if (params.version) { @@ -62,14 +33,6 @@ class WorkflowMain { System.exit(0) } - // Print parameter summary log to screen - log.info paramsSummaryLog(workflow, params) - - // Validate workflow parameters via the JSON schema - if (params.validate_params) { - NfcoreSchema.validateParameters(workflow, params, log) - } - // Check that a -profile or Nextflow config has been provided to run the pipeline NfcoreTemplate.checkConfigProvided(workflow, log) diff --git a/main.nf b/main.nf index 2071b49a..c5573a01 100644 --- a/main.nf +++ b/main.nf @@ -25,6 +25,22 @@ params.fasta = WorkflowMain.getGenomeAttribute(params, 'fasta') ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ +include { validateParameters; paramsHelp } from 'plugin/nf-validation' + +// Print help message if needed +if (params.help) { + def logo = NfcoreTemplate.logo(workflow, params.monochrome_logs) + def citation = '\n' + WorkflowMain.citation(workflow) + '\n' + def String command = "nextflow run ${workflow.manifest.name} --input samplesheet.csv --genome GRCh37 -profile docker" + log.info logo + paramsHelp(command) + citation + NfcoreTemplate.dashedLine(params.monochrome_logs) + System.exit(0) +} + +// Validate input parameters +if (params.validate_params) { + validateParameters() +} + WorkflowMain.initialise(workflow, params, log) /* diff --git a/nextflow.config b/nextflow.config index ae9ed185..c7eaca03 100644 --- a/nextflow.config +++ b/nextflow.config @@ -12,12 +12,12 @@ params { // TODO nf-core: Specify your pipeline's command line flags // Input options input = null - - // References genome = null igenomes_base = 's3://ngi-igenomes/igenomes' igenomes_ignore = false + + // MultiQC options multiqc_config = null multiqc_title = null @@ -27,7 +27,6 @@ params { // Boilerplate options outdir = null - tracedir = "${params.outdir}/pipeline_info" publish_dir_mode = 'copy' email = null email_on_fail = null @@ -36,19 +35,15 @@ params { hook_url = null help = false version = false - validate_params = true - show_hidden_params = false - schema_ignore_params = 'genomes' - // Config options + config_profile_name = null + config_profile_description = null custom_config_version = 'master' custom_config_base = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}" - config_profile_description = null config_profile_contact = null config_profile_url = null - config_profile_name = null - + // Max resource options // Defaults only, expecting to be overwritten @@ -56,6 +51,13 @@ params { max_cpus = 16 max_time = '240.h' + // Schema validation default options + validationFailUnrecognisedParams = false + validationLenientMode = false + validationSchemaIgnoreParams = 'genomes' + validationShowHiddenParams = false + validate_params = true + } // Load base.config by default for all pipelines @@ -75,13 +77,11 @@ try { // } catch (Exception e) { // System.err.println("WARNING: Could not load nf-core/config/crisprseq profiles: ${params.custom_config_base}/pipeline/crisprseq.config") // } - - profiles { debug { dumpHashes = true process.beforeScript = 'echo $HOSTNAME' - cleanup = false + cleanup = false } conda { conda.enabled = true @@ -104,7 +104,6 @@ profiles { } docker { docker.enabled = true - docker.registry = 'quay.io' docker.userEmulation = true conda.enabled = false singularity.enabled = false @@ -128,7 +127,6 @@ profiles { } podman { podman.enabled = true - podman.registry = 'quay.io' conda.enabled = false docker.enabled = false singularity.enabled = false @@ -172,6 +170,18 @@ profiles { test_full { includeConfig 'conf/test_full.config' } } +// Set default registry for Apptainer, Docker, Podman and Singularity independent of -profile +// Will not be used unless Apptainer / Docker / Podman / Singularity are enabled +// Set to your registry if you have a mirror of containers +apptainer.registry = 'quay.io' +docker.registry = 'quay.io' +podman.registry = 'quay.io' +singularity.registry = 'quay.io' + +// Nextflow plugins +plugins { + id 'nf-validation' // Validation of pipeline parameters and creation of an input channel from a sample sheet +} // Load igenomes.config if required if (!params.igenomes_ignore) { @@ -179,8 +189,6 @@ if (!params.igenomes_ignore) { } else { params.genomes = [:] } - - // Export these variables to prevent local Python/R libraries from conflicting with those in the container // The JULIA depot path has been adjusted to a fixed path `/usr/local/share/julia` that needs to be used for packages in the container. // See https://apeltzer.github.io/post/03-julia-lang-nextflow/ for details on that. Once we have a common agreement on where to keep Julia packages, this is adjustable. @@ -198,19 +206,19 @@ process.shell = ['/bin/bash', '-euo', 'pipefail'] def trace_timestamp = new java.util.Date().format( 'yyyy-MM-dd_HH-mm-ss') timeline { enabled = true - file = "${params.tracedir}/execution_timeline_${trace_timestamp}.html" + file = "${params.outdir}/pipeline_info/execution_timeline_${trace_timestamp}.html" } report { enabled = true - file = "${params.tracedir}/execution_report_${trace_timestamp}.html" + file = "${params.outdir}/pipeline_info/execution_report_${trace_timestamp}.html" } trace { enabled = true - file = "${params.tracedir}/execution_trace_${trace_timestamp}.txt" + file = "${params.outdir}/pipeline_info/execution_trace_${trace_timestamp}.txt" } dag { enabled = true - file = "${params.tracedir}/pipeline_dag_${trace_timestamp}.html" + file = "${params.outdir}/pipeline_info/pipeline_dag_${trace_timestamp}.html" } manifest { @@ -219,8 +227,8 @@ manifest { homePage = 'https://github.com/nf-core/crisprseq' description = """Pipeline for the analysis of crispr data""" mainScript = 'main.nf' - nextflowVersion = '!>=22.10.1' - version = '1.0' + nextflowVersion = '!>=23.04.0' + version = '2.0.0' doi = '' } diff --git a/nextflow_schema.json b/nextflow_schema.json index 0699e01c..815e1887 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -15,9 +15,9 @@ "input": { "type": "string", "format": "file-path", + "exists": true, "mimetype": "text/csv", "pattern": "^\\S+\\.csv$", - "schema": "assets/schema_input.json", "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/crisprseq/usage#samplesheet-input).", "fa_icon": "fas fa-file-csv" @@ -57,6 +57,7 @@ "fasta": { "type": "string", "format": "file-path", + "exists": true, "mimetype": "text/plain", "pattern": "^\\S+\\.fn?a(sta)?(\\.gz)?$", "description": "Path to FASTA genome file.", @@ -157,7 +158,7 @@ "description": "Maximum amount of time that can be requested for any single job.", "default": "240.h", "fa_icon": "far fa-clock", - "pattern": "^(\\d+\\.?\\s*(s|m|h|day)\\s*)+$", + "pattern": "^(\\d+\\.?\\s*(s|m|h|d|day)\\s*)+$", "hidden": true, "help_text": "Use to set an upper-limit for the time requirement for each process. Should be a string in the format integer-unit e.g. `--max_time '2.h'`" } @@ -174,12 +175,14 @@ "type": "boolean", "description": "Display help text.", "fa_icon": "fas fa-question-circle", + "default": false, "hidden": true }, "version": { "type": "boolean", "description": "Display version and exit.", "fa_icon": "fas fa-question-circle", + "default": false, "hidden": true }, "publish_dir_mode": { @@ -203,6 +206,7 @@ "type": "boolean", "description": "Send plain-text email instead of HTML.", "fa_icon": "fas fa-remove-format", + "default": false, "hidden": true }, "max_multiqc_email_size": { @@ -217,6 +221,7 @@ "type": "boolean", "description": "Do not use coloured log outputs.", "fa_icon": "fas fa-palette", + "default": false, "hidden": true }, "hook_url": { @@ -228,6 +233,7 @@ }, "multiqc_config": { "type": "string", + "format": "file-path", "description": "Custom config file to supply to MultiQC.", "fa_icon": "fas fa-cog", "hidden": true @@ -243,13 +249,6 @@ "description": "Custom MultiQC yaml file containing HTML including a methods description.", "fa_icon": "fas fa-cog" }, - "tracedir": { - "type": "string", - "description": "Directory to keep pipeline Nextflow logs and reports.", - "default": "${params.outdir}/pipeline_info", - "fa_icon": "fas fa-cogs", - "hidden": true - }, "validate_params": { "type": "boolean", "description": "Boolean whether to validate parameters against the schema at runtime", @@ -257,12 +256,29 @@ "fa_icon": "fas fa-check-square", "hidden": true }, - "show_hidden_params": { + "validationShowHiddenParams": { "type": "boolean", "fa_icon": "far fa-eye-slash", "description": "Show all params when using `--help`", + "default": false, "hidden": true, "help_text": "By default, parameters set as _hidden_ in the schema are not shown on the command line when a user runs with `--help`. Specifying this option will tell the pipeline to show all parameters." + }, + "validationFailUnrecognisedParams": { + "type": "boolean", + "fa_icon": "far fa-check-circle", + "description": "Validation of parameters fails when an unrecognised parameter is found.", + "default": false, + "hidden": true, + "help_text": "By default, when an unrecognised parameter is found, it returns a warinig." + }, + "validationLenientMode": { + "type": "boolean", + "fa_icon": "far fa-check-circle", + "description": "Validation of parameters in lenient more.", + "default": false, + "hidden": true, + "help_text": "Allows string values that are parseable as numbers or booleans. For further information see [JSONSchema docs](https://github.com/everit-org/json-schema#lenient-mode)." } } } diff --git a/workflows/crisprseq.nf b/workflows/crisprseq.nf index 91a0baf8..d608ede8 100644 --- a/workflows/crisprseq.nf +++ b/workflows/crisprseq.nf @@ -1,21 +1,19 @@ /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - VALIDATE INPUTS + PRINT PARAMS SUMMARY ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -def summary_params = NfcoreSchema.paramsSummaryMap(workflow, params) +include { paramsSummaryLog; paramsSummaryMap } from 'plugin/nf-validation' -// Validate input parameters -WorkflowCrisprseq.initialise(params, log) +def logo = NfcoreTemplate.logo(workflow, params.monochrome_logs) +def citation = '\n' + WorkflowMain.citation(workflow) + '\n' +def summary_params = paramsSummaryMap(workflow) -// TODO nf-core: Add all file path parameters for the pipeline to the list below -// Check input path parameters to see if they exist -def checkPathParamList = [ params.input, params.multiqc_config, params.fasta ] -for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } } +// Print parameter summary log to screen +log.info logo + paramsSummaryLog(workflow) + citation -// Check mandatory parameters -if (params.input) { ch_input = file(params.input) } else { exit 1, 'Input samplesheet not specified!' } +WorkflowCrisprseq.initialise(params, log) /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -69,9 +67,12 @@ workflow CRISPRSEQ { // SUBWORKFLOW: Read in samplesheet, validate and stage input files // INPUT_CHECK ( - ch_input + file(params.input) ) ch_versions = ch_versions.mix(INPUT_CHECK.out.versions) + // TODO: OPTIONAL, you can use nf-validation plugin to create an input channel from the samplesheet with Channel.fromSamplesheet("input") + // See the documentation https://nextflow-io.github.io/nf-validation/samplesheets/fromSamplesheet/ + // ! There is currently no tooling to help you write a sample sheet schema // // MODULE: Run FastQC @@ -91,7 +92,7 @@ workflow CRISPRSEQ { workflow_summary = WorkflowCrisprseq.paramsSummaryMultiqc(workflow, summary_params) ch_workflow_summary = Channel.value(workflow_summary) - methods_description = WorkflowCrisprseq.methodsDescriptionText(workflow, ch_multiqc_custom_methods_description) + methods_description = WorkflowCrisprseq.methodsDescriptionText(workflow, ch_multiqc_custom_methods_description, params) ch_methods_description = Channel.value(methods_description) ch_multiqc_files = Channel.empty() From c006f62a4631ba0c5794f3ee3e80019c22cd3398 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 5 Jul 2023 10:26:26 +0200 Subject: [PATCH 002/133] Bump dev version --- CHANGELOG.md | 8 ++++++++ README.md | 4 ++-- lib/WorkflowMain.groovy | 2 +- nextflow.config | 8 ++++---- 4 files changed, 15 insertions(+), 7 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 1854e1fa..d1ddf4f6 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,6 +3,14 @@ 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). +## [v2.1.0dev] + +### Added + +### Fixed + +### Deprecated + ## [v2.0.0 - Paprika Lovelace](https://github.com/nf-core/crisprseq/releases/tag/2.0.0) - [05.07.2023] ### Added diff --git a/README.md b/README.md index 046675c7..41ce5924 100644 --- a/README.md +++ b/README.md @@ -1,6 +1,6 @@ # ![nf-core/crisprseq](docs/images/nf-core-crisprseq_logo_light.png#gh-light-mode-only) ![nf-core/crisprseq](docs/images/nf-core-crisprseq_logo_dark.png#gh-dark-mode-only) -[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.7598497-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.7598497) +[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.8116072-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.8116072) [![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A523.04.0-23aa62.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/) @@ -126,7 +126,7 @@ For further information or help, don't hesitate to get in touch on the [Slack `# ## Citations -If you use nf-core/crisprseq for your analysis, please cite it using the following doi: [10.5281/zenodo.7598497](https://doi.org/10.5281/zenodo.7598497) +If you use nf-core/crisprseq for your analysis, please cite it using the following doi: [10.5281/zenodo.8116072](https://doi.org/10.5281/zenodo.8116072) An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file. diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy index 05c17b7f..7fa1e0e1 100755 --- a/lib/WorkflowMain.groovy +++ b/lib/WorkflowMain.groovy @@ -12,7 +12,7 @@ class WorkflowMain { public static String citation(workflow) { return "If you use ${workflow.manifest.name} for your analysis please cite:\n\n" + "* The pipeline\n" + - " https://doi.org/10.5281/zenodo.7598497\n\n" + + " https://doi.org/10.5281/zenodo.8116072\n\n" + "* The nf-core framework\n" + " https://doi.org/10.1038/s41587-020-0439-x\n\n" + "* Software dependencies\n" + diff --git a/nextflow.config b/nextflow.config index 05bda0dd..410e8c7c 100644 --- a/nextflow.config +++ b/nextflow.config @@ -247,13 +247,13 @@ dag { manifest { name = 'nf-core/crisprseq' - author = """mirpedrol""" + author = """Júlia Mir Pedrol, Laurence Kuhlburger""" homePage = 'https://github.com/nf-core/crisprseq' - description = """Pipeline for the analysis of crispr data""" + description = """Pipeline for the analysis of CRISPR data""" mainScript = 'main.nf' nextflowVersion = '!>=23.04.0' - version = '2.0.0' - doi = 'https://doi.org/10.5281/zenodo.7598497' + version = '2.1.0dev' + doi = 'https://doi.org/10.5281/zenodo.8116072' } // Load modules.config for DSL2 module specific options From f1dc329df1b80a272db4ae557721a921d96f713a Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 10 Jul 2023 11:38:16 +0200 Subject: [PATCH 003/133] fix linting from template --- LICENSE | 2 +- assets/email_template.html | 2 +- assets/multiqc_config.yml | 2 +- nextflow_schema.json | 2 +- 4 files changed, 4 insertions(+), 4 deletions(-) diff --git a/LICENSE b/LICENSE index 41427d4d..4de6f708 100644 --- a/LICENSE +++ b/LICENSE @@ -1,6 +1,6 @@ MIT License -Copyright (c) mirpedrol +Copyright (c) Júlia Mir Pedrol, Laurence Kuhlburger 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/assets/email_template.html b/assets/email_template.html index 4a90e11b..ec162f42 100644 --- a/assets/email_template.html +++ b/assets/email_template.html @@ -4,7 +4,7 @@ - + nf-core/crisprseq Pipeline Report diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 5f5e0401..67439316 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -1,5 +1,5 @@ report_comment: > - This report has been generated by the nf-core/crisprseq + This report has been generated by the nf-core/crisprseq analysis pipeline. For information about how to interpret these results, please see the documentation. report_section_order: diff --git a/nextflow_schema.json b/nextflow_schema.json index 5d81138d..ffef1b81 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -2,7 +2,7 @@ "$schema": "http://json-schema.org/draft-07/schema", "$id": "https://raw.githubusercontent.com/nf-core/crisprseq/master/nextflow_schema.json", "title": "nf-core/crisprseq pipeline parameters", - "description": "Pipeline for the analysis of crispr data", + "description": "Pipeline for the analysis of CRISPR data", "type": "object", "definitions": { "input_output_options": { From 8297a8129555c17a58d105c55c9ee875d026ddd4 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 10 Jul 2023 16:10:17 +0200 Subject: [PATCH 004/133] fix duplicated format in json schema --- nextflow_schema.json | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index ffef1b81..96005eae 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -201,8 +201,7 @@ "mimetype": "text/plain", "pattern": "^\\S+\\.fn?a(sta)?(\\.gz)?$", "description": "Path to the reference FASTA file. Will override reference sequences provided by an input sample sheet.", - "fa_icon": "far fa-file-alt", - "format": "file-path" + "fa_icon": "far fa-file-alt" }, "igenomes_base": { "type": "string", From bca2bdf58bf3fd33cc20fdced79fc3afcc4e3da0 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 13 Jul 2023 09:53:30 +0200 Subject: [PATCH 005/133] validate params through the schema --- nextflow_schema.json | 15 ++++++++------- workflows/crisprseq_screening.nf | 18 ++++++++---------- 2 files changed, 16 insertions(+), 17 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 96005eae..a5be54ca 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -151,18 +151,26 @@ "properties": { "mle_design_matrix": { "type": "string", + "format": "file-path", + "exists": true, "description": "Design matrix used for MAGeCK MLE to call essential genes under multiple conditions while considering sgRNA knockout efficiency" }, "rra_contrasts": { "type": "string", + "format": "file-path", + "exists": true, "description": "Comma-separated file with the conditions to be compared. The first one will be the reference (control)" }, "count_table": { "type": "string", + "format": "file-path", + "exists": true, "description": "Please provide your count table if the mageck test should be skipped." }, "library": { "type": "string", + "format": "file-path", + "exists": true, "fa_icon": "far fa-address-book", "description": "sgRNA sequences and targeting genes" }, @@ -314,14 +322,12 @@ "type": "boolean", "description": "Display help text.", "fa_icon": "fas fa-question-circle", - "default": false, "hidden": true }, "version": { "type": "boolean", "description": "Display version and exit.", "fa_icon": "fas fa-question-circle", - "default": false, "hidden": true }, "publish_dir_mode": { @@ -345,7 +351,6 @@ "type": "boolean", "description": "Send plain-text email instead of HTML.", "fa_icon": "fas fa-remove-format", - "default": false, "hidden": true }, "max_multiqc_email_size": { @@ -360,7 +365,6 @@ "type": "boolean", "description": "Do not use coloured log outputs.", "fa_icon": "fas fa-palette", - "default": false, "hidden": true }, "hook_url": { @@ -399,7 +403,6 @@ "type": "boolean", "fa_icon": "far fa-eye-slash", "description": "Show all params when using `--help`", - "default": false, "hidden": true, "help_text": "By default, parameters set as _hidden_ in the schema are not shown on the command line when a user runs with `--help`. Specifying this option will tell the pipeline to show all parameters." }, @@ -414,7 +417,6 @@ "type": "boolean", "fa_icon": "far fa-check-circle", "description": "Validation of parameters fails when an unrecognised parameter is found.", - "default": false, "hidden": true, "help_text": "By default, when an unrecognised parameter is found, it returns a warinig." }, @@ -422,7 +424,6 @@ "type": "boolean", "fa_icon": "far fa-check-circle", "description": "Validation of parameters in lenient more.", - "default": false, "hidden": true, "help_text": "Allows string values that are parseable as numbers or booleans. For further information see [JSONSchema docs](https://github.com/everit-org/json-schema#lenient-mode)." } diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index ec989f40..b9ed6237 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -15,16 +15,14 @@ log.info logo + paramsSummaryLog(workflow) + citation WorkflowCrisprseq.initialise(params, log) -// Check mandatory parameters -//if (!params.count_table) { ch_input = file(params.input) } else { error('Input samplesheet not specified!') } -//if (params.library) { ch_library = file(params.library) } -//if (params.crisprcleanr) { ch_crisprcleanr= Channel.value(params.crisprcleanr) } - -//if(params.mle_design_matrix) { -// Channel.fromPath(params.mle_design_matrix,checkIfExists: true) -// .set { ch_design } -//} -// TODO check with the schema +// Set screening parameters and channels +if (params.library) { ch_library = file(params.library) } +if (params.crisprcleanr) { ch_crisprcleanr = Channel.value(params.crisprcleanr) } + +if(params.mle_design_matrix) { + Channel.fromPath(params.mle_design_matrix) + .set { ch_design } +} /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ From f29d9469a683ea250434ce868d39a8882d8c648d Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 13 Jul 2023 09:54:52 +0200 Subject: [PATCH 006/133] update changelog --- CHANGELOG.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index d1ddf4f6..02d741b5 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -7,6 +7,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Added +- Template update v2.9 ([#52](https://github.com/nf-core/crisprseq/pull/52)) + ### Fixed ### Deprecated From e998e17f618614e17acbf664d5d1dc5dada98527 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 13 Jul 2023 10:15:14 +0200 Subject: [PATCH 007/133] fix link in multipqc_config.yml --- assets/multiqc_config.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 67439316..73d174a7 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -1,7 +1,7 @@ report_comment: > This report has been generated by the nf-core/crisprseq analysis pipeline. For information about how to interpret these results, please see the - documentation. + documentation. report_section_order: "nf-core-crisprseq-methods-description": order: -1000 From c73dbfb287ed56c1eba9b83d706014936f358ddb Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Fri, 14 Jul 2023 08:51:08 +0200 Subject: [PATCH 008/133] Update nextflow_schema.json --- nextflow_schema.json | 6 ++++-- 1 file changed, 4 insertions(+), 2 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index a5be54ca..36d4cd2a 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -164,15 +164,17 @@ "count_table": { "type": "string", "format": "file-path", + "mimetype": "text/tsv", "exists": true, "description": "Please provide your count table if the mageck test should be skipped." }, "library": { "type": "string", "format": "file-path", + "mimetype": "text/tsv", "exists": true, "fa_icon": "far fa-address-book", - "description": "sgRNA sequences and targeting genes" + "description": "sgRNA and targetting genes, tab separated" }, "crisprcleanr": { "type": "string", @@ -185,7 +187,7 @@ }, "min_targeted_genes": { "type": "number", - "description": "Minimal number of different genes targeted by sgRNAs in a biased segment in order for the corresponding counts to be corrected", + "description": "Minimal number of different genes targeted by sgRNAs in a biased segment in order for the corresponding counts to be corrected for CRISPRcleanR", "default": 3 } } From 28665e083fc4a5a5b20ba97eefdb22cc76d74e1a Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C3=BAlia=20Mir=20Pedrol?= Date: Fri, 14 Jul 2023 08:54:12 +0200 Subject: [PATCH 009/133] Apply suggestions from code review --- nextflow_schema.json | 2 ++ 1 file changed, 2 insertions(+) diff --git a/nextflow_schema.json b/nextflow_schema.json index 36d4cd2a..48f8656c 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -164,6 +164,7 @@ "count_table": { "type": "string", "format": "file-path", + "pattern": "^\\S+\\.tsv$", "mimetype": "text/tsv", "exists": true, "description": "Please provide your count table if the mageck test should be skipped." @@ -171,6 +172,7 @@ "library": { "type": "string", "format": "file-path", + "pattern": "^\\S+\\.csv$", "mimetype": "text/tsv", "exists": true, "fa_icon": "far fa-address-book", From ef705d7c1fdd44565dc6f89b8434bdf893d64d86 Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C3=BAlia=20Mir=20Pedrol?= Date: Fri, 14 Jul 2023 09:51:43 +0200 Subject: [PATCH 010/133] Apply suggestions from code review Co-authored-by: LaurenceKuhl --- nextflow_schema.json | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 48f8656c..ce0956c1 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -164,7 +164,7 @@ "count_table": { "type": "string", "format": "file-path", - "pattern": "^\\S+\\.tsv$", + "pattern": "^\\S+\\.(tsv|txt)$", "mimetype": "text/tsv", "exists": true, "description": "Please provide your count table if the mageck test should be skipped." @@ -172,7 +172,7 @@ "library": { "type": "string", "format": "file-path", - "pattern": "^\\S+\\.csv$", + "pattern": "^\\S+\\.(tsv|txt)$", "mimetype": "text/tsv", "exists": true, "fa_icon": "far fa-address-book", From 8f6af2d596b8fffdb94a67c05077d9ce618888d0 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Fri, 14 Jul 2023 11:00:01 +0200 Subject: [PATCH 011/133] Extra files from mageck count --- modules/nf-core/mageck/count/mageck-count.diff | 9 ++++++--- modules/nf-core/mageck/count/main.nf | 3 +++ 2 files changed, 9 insertions(+), 3 deletions(-) diff --git a/modules/nf-core/mageck/count/mageck-count.diff b/modules/nf-core/mageck/count/mageck-count.diff index 50ee78b1..48cd0c4f 100644 --- a/modules/nf-core/mageck/count/mageck-count.diff +++ b/modules/nf-core/mageck/count/mageck-count.diff @@ -1,24 +1,27 @@ Changes in module 'nf-core/mageck/count' --- modules/nf-core/mageck/count/main.nf +++ modules/nf-core/mageck/count/main.nf -@@ -12,8 +12,10 @@ +@@ -12,8 +12,13 @@ path(library) output: - tuple val(meta), path("*count*.txt"), emit: count + tuple val(meta), path("*count.txt"), emit: count tuple val(meta), path("*.count_normalized.txt"), emit: norm ++ tuple val(meta), path("*.countsummary.txt"), emit: summary ++ tuple val(meta), path("*.count_normalized.txt"), emit: normalized ++ tuple val(meta), path("*.count_report.Rmd"), emit: report + tuple val(meta), path("*.log"), emit: logs + path "versions.yml" , emit: versions when: -@@ -22,7 +24,7 @@ +@@ -22,7 +27,7 @@ script: def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" - def input_file = ("$inputfile".endsWith(".fastq.gz")) ? "--fastq ${inputfile}" : "-k ${inputfile}" -+ def input_file = ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz")) ? "--fastq ${inputfile}" : "-k ${inputfile}" ++ def input_file = ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz")) ? "--fastq ${inputfile}" : "-k ${inputfile}" def sample_label = ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz")) ? "--sample-label ${meta.id}" : '' """ diff --git a/modules/nf-core/mageck/count/main.nf b/modules/nf-core/mageck/count/main.nf index 30fbd8fe..91ea3fe2 100644 --- a/modules/nf-core/mageck/count/main.nf +++ b/modules/nf-core/mageck/count/main.nf @@ -14,6 +14,9 @@ process MAGECK_COUNT { output: tuple val(meta), path("*count.txt"), emit: count tuple val(meta), path("*.count_normalized.txt"), emit: norm + tuple val(meta), path("*.countsummary.txt"), emit: summary + tuple val(meta), path("*.count_normalized.txt"), emit: normalized + tuple val(meta), path("*.count_report.Rmd"), emit: report tuple val(meta), path("*.log"), emit: logs path "versions.yml" , emit: versions From 13181f093fe4b45b90c324f484c857d5e593424d Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Fri, 14 Jul 2023 14:17:02 +0200 Subject: [PATCH 012/133] fix bug in rra --- modules/nf-core/mageck/count/mageck-count.diff | 5 ++--- modules/nf-core/mageck/count/main.nf | 1 - modules/nf-core/mageck/test/mageck-test.diff | 12 +++--------- modules/nf-core/mageck/test/main.nf | 1 - 4 files changed, 5 insertions(+), 14 deletions(-) diff --git a/modules/nf-core/mageck/count/mageck-count.diff b/modules/nf-core/mageck/count/mageck-count.diff index 48cd0c4f..87f8e83c 100644 --- a/modules/nf-core/mageck/count/mageck-count.diff +++ b/modules/nf-core/mageck/count/mageck-count.diff @@ -1,7 +1,7 @@ Changes in module 'nf-core/mageck/count' --- modules/nf-core/mageck/count/main.nf +++ modules/nf-core/mageck/count/main.nf -@@ -12,8 +12,13 @@ +@@ -12,8 +12,12 @@ path(library) output: @@ -10,13 +10,12 @@ Changes in module 'nf-core/mageck/count' tuple val(meta), path("*.count_normalized.txt"), emit: norm + tuple val(meta), path("*.countsummary.txt"), emit: summary + tuple val(meta), path("*.count_normalized.txt"), emit: normalized -+ tuple val(meta), path("*.count_report.Rmd"), emit: report + tuple val(meta), path("*.log"), emit: logs + path "versions.yml" , emit: versions when: -@@ -22,7 +27,7 @@ +@@ -22,7 +26,7 @@ script: def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" diff --git a/modules/nf-core/mageck/count/main.nf b/modules/nf-core/mageck/count/main.nf index 91ea3fe2..01691029 100644 --- a/modules/nf-core/mageck/count/main.nf +++ b/modules/nf-core/mageck/count/main.nf @@ -16,7 +16,6 @@ process MAGECK_COUNT { tuple val(meta), path("*.count_normalized.txt"), emit: norm tuple val(meta), path("*.countsummary.txt"), emit: summary tuple val(meta), path("*.count_normalized.txt"), emit: normalized - tuple val(meta), path("*.count_report.Rmd"), emit: report tuple val(meta), path("*.log"), emit: logs path "versions.yml" , emit: versions diff --git a/modules/nf-core/mageck/test/mageck-test.diff b/modules/nf-core/mageck/test/mageck-test.diff index ee8558c3..9da13d3b 100644 --- a/modules/nf-core/mageck/test/mageck-test.diff +++ b/modules/nf-core/mageck/test/mageck-test.diff @@ -8,17 +8,16 @@ Changes in module 'nf-core/mageck/test' label 'process_medium' conda "bioconda::mageck=0.5.9" -@@ -14,6 +14,9 @@ +@@ -14,6 +14,8 @@ tuple val(meta), path("*.gene_summary.txt") , emit: gene_summary tuple val(meta), path("*.sgrna_summary.txt") , emit: sgrna_summary tuple val(meta), path("*.R") , emit: r_script -+ tuple val(meta), path("*.Rmd") , emit: r_report + tuple val(meta), path("*.Rnw") , emit: r_summary + tuple val(meta), path("*.log") , emit: logs path "versions.yml" , emit: versions when: -@@ -21,14 +24,18 @@ +@@ -21,14 +23,18 @@ script: def args = task.ext.args ?: '' @@ -38,10 +37,5 @@ Changes in module 'nf-core/mageck/test' cat <<-END_VERSIONS > versions.yml -@@ -36,4 +43,4 @@ - mageck: \$(mageck -v) - END_VERSIONS - """ --} -+} + ************************************************************ diff --git a/modules/nf-core/mageck/test/main.nf b/modules/nf-core/mageck/test/main.nf index 420edcc1..78d5f58c 100644 --- a/modules/nf-core/mageck/test/main.nf +++ b/modules/nf-core/mageck/test/main.nf @@ -14,7 +14,6 @@ process MAGECK_TEST { tuple val(meta), path("*.gene_summary.txt") , emit: gene_summary tuple val(meta), path("*.sgrna_summary.txt") , emit: sgrna_summary tuple val(meta), path("*.R") , emit: r_script - tuple val(meta), path("*.Rmd") , emit: r_report tuple val(meta), path("*.Rnw") , emit: r_summary tuple val(meta), path("*.log") , emit: logs path "versions.yml" , emit: versions From 2226e704d8e3a9f6c6b8c3c3795bf070bf677331 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 18 Jul 2023 15:24:29 +0200 Subject: [PATCH 013/133] add fromSamplesheet() in targete dpipeline --- assets/schema_input.json | 31 +++++++++++++-- conf/modules.config | 8 ---- lib/WorkflowCrisprseq.groovy | 27 +++++++++++++ lib/WorkflowMain.groovy | 5 --- nextflow_schema.json | 6 +++ workflows/crisprseq_screening.nf | 9 ++--- workflows/crisprseq_targeted.nf | 66 +++++++++++++++++--------------- 7 files changed, 101 insertions(+), 51 deletions(-) diff --git a/assets/schema_input.json b/assets/schema_input.json index 7327787f..a66228ed 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -10,25 +10,50 @@ "sample": { "type": "string", "pattern": "^\\S+$", - "errorMessage": "Sample name must be provided and cannot contain spaces" + "errorMessage": "Sample name must be provided and cannot contain spaces", + "meta": ["id"] }, "fastq_1": { "type": "string", + "format": "file-path", + "exists": true, "pattern": "^\\S+\\.f(ast)?q\\.gz$", "errorMessage": "FastQ file for reads 1 must be provided, cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'" }, "fastq_2": { + "type": "string", + "format": "file-path", + "exists": true, "errorMessage": "FastQ file for reads 2 cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'", "anyOf": [ { - "type": "string", "pattern": "^\\S+\\.f(ast)?q\\.gz$" }, { - "type": "string", "maxLength": 0 } ] + }, + "condition": { + "type": "string", + "pattern": "^\\S+$", + "enum": ["control", "treatment"], + "errorMessage": "Condition name must be one of 'control' or 'treatment'" + }, + "reference": { + "type": "string", + "pattern": "^[ACTGN]+$", + "errorMessage": "Reference sequence must be a valid DNA sequence" + }, + "protospacer": { + "type": "string", + "pattern": "^[ACTGN]+$", + "errorMessage": "Protospacer sequence must be a valid DNA sequence" + }, + "template": { + "type": "string", + "pattern": "^[ACTGN]+$", + "errorMessage": "Template sequence must be a valid DNA sequence" } }, "required": ["sample", "fastq_1"] diff --git a/conf/modules.config b/conf/modules.config index 84504a31..43cdc492 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -18,14 +18,6 @@ process { saveAs: { filename -> filename.equals('versions.yml') ? null : filename } ] - withName: SAMPLESHEET_CHECK { - publishDir = [ - path: { "${params.outdir}/pipeline_info" }, - mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } - ] - } - withName: SEQ_TO_FILE_REF { publishDir = [ path: { "${params.outdir}/preprocessing/sequences" }, diff --git a/lib/WorkflowCrisprseq.groovy b/lib/WorkflowCrisprseq.groovy index 85035344..289319ff 100755 --- a/lib/WorkflowCrisprseq.groovy +++ b/lib/WorkflowCrisprseq.groovy @@ -15,6 +15,33 @@ class WorkflowCrisprseq { genomeExistsError(params, log) } + // + // Function to validate channels from input samplesheet + // + public static ArrayList validateInput(input) { + def (metas, fastqs) = input[1..2] + + // Check that multiple runs of the same sample are of the same datatype i.e. single-end / paired-end + def endedness_ok = metas.collect{ it.single_end }.unique().size == 1 + if (!endedness_ok) { + Nextflow.error("Please check input samplesheet -> Multiple runs of a sample must be of the same datatype i.e. single-end or paired-end: ${metas[0].id}") + } + + // Check that multiple runs of the same sample contain a reference or not + def reference_ok = metas.collect{ it.self_reference }.unique().size == 1 + if (!reference_ok) { + Nextflow.error("Please check input samplesheet -> Multiple runs of a sample must all contain a reference or not: ${metas[0].id}") + } + + // Check that multiple runs of the same sample contain a template or not + def template_ok = metas.collect{ it.template }.unique().size == 1 + if (!template_ok) { + Nextflow.error("Please check input samplesheet -> Multiple runs of a sample must all contain a template or not: ${metas[0].id}") + } + + return [ metas[0], fastqs ] + } + // // Get workflow summary for MultiQC // diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy index 8f725734..36db450c 100755 --- a/lib/WorkflowMain.groovy +++ b/lib/WorkflowMain.groovy @@ -42,11 +42,6 @@ class WorkflowMain { // Check AWS batch settings NfcoreTemplate.awsBatch(workflow, params) - - // Check input has been provided - if (!params.input) { - Nextflow.error("Please provide an input samplesheet to the pipeline e.g. '--input samplesheet.csv'") - } } // // Get attribute from genome config file e.g. fasta diff --git a/nextflow_schema.json b/nextflow_schema.json index ce0956c1..c75c5a2d 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -16,6 +16,7 @@ "type": "string", "format": "file-path", "exists": true, + "schema": "assets/schema_input.json", "mimetype": "text/csv", "pattern": "^\\S+\\.csv$", "description": "Path to comma-separated file containing information about the samples in the experiment.", @@ -381,18 +382,23 @@ "multiqc_config": { "type": "string", "format": "file-path", + "exists": true, "description": "Custom config file to supply to MultiQC.", "fa_icon": "fas fa-cog", "hidden": true }, "multiqc_logo": { "type": "string", + "format": "file-path", + "exists": true, "description": "Custom logo file to supply to MultiQC. File name must also be set in the MultiQC config file", "fa_icon": "fas fa-image", "hidden": true }, "multiqc_methods_description": { "type": "string", + "format": "file-path", + "exists": true, "description": "Custom MultiQC yaml file containing HTML including a methods description.", "fa_icon": "fas fa-cog" }, diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index b9ed6237..ff6fcada 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -4,7 +4,7 @@ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -include { paramsSummaryLog; paramsSummaryMap } from 'plugin/nf-validation' +include { paramsSummaryLog; paramsSummaryMap; fromSamplesheet } from 'plugin/nf-validation' def logo = NfcoreTemplate.logo(workflow, params.monochrome_logs) def citation = '\n' + WorkflowMain.citation(workflow) + '\n' @@ -31,9 +31,9 @@ if(params.mle_design_matrix) { */ ch_multiqc_config = Channel.fromPath("$projectDir/assets/multiqc_config.yml", checkIfExists: true) -ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath( params.multiqc_config, checkIfExists: true ) : Channel.empty() -ch_multiqc_logo = params.multiqc_logo ? Channel.fromPath( params.multiqc_logo, checkIfExists: true ) : Channel.empty() -ch_multiqc_custom_methods_description = params.multiqc_methods_description ? file(params.multiqc_methods_description, checkIfExists: true) : file("$projectDir/assets/methods_description_template.yml", checkIfExists: true) +ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath( params.multiqc_config ) : Channel.empty() +ch_multiqc_logo = params.multiqc_logo ? Channel.fromPath( params.multiqc_logo ) : Channel.empty() +ch_multiqc_custom_methods_description = params.multiqc_methods_description ? file(params.multiqc_methods_description) : file("$projectDir/assets/methods_description_template.yml", checkIfExists: true) /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -44,7 +44,6 @@ ch_multiqc_custom_methods_description = params.multiqc_methods_description ? fil // // SUBWORKFLOW: Consisting of a mix of local and nf-core/modules // -include { INPUT_CHECK_SCREENING } from '../subworkflows/local/input_check_screening' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index ff14ff28..8f4d33f7 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -4,7 +4,7 @@ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ -include { paramsSummaryLog; paramsSummaryMap } from 'plugin/nf-validation' +include { paramsSummaryLog; paramsSummaryMap; fromSamplesheet } from 'plugin/nf-validation' def logo = NfcoreTemplate.logo(workflow, params.monochrome_logs) def citation = '\n' + WorkflowMain.citation(workflow) + '\n' @@ -21,9 +21,9 @@ WorkflowCrisprseq.initialise(params, log) */ ch_multiqc_config = Channel.fromPath("$projectDir/assets/multiqc_config.yml", checkIfExists: true) -ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath( params.multiqc_config, checkIfExists: true ) : Channel.empty() -ch_multiqc_logo = params.multiqc_logo ? Channel.fromPath( params.multiqc_logo, checkIfExists: true ) : Channel.empty() -ch_multiqc_custom_methods_description = params.multiqc_methods_description ? file(params.multiqc_methods_description, checkIfExists: true) : file("$projectDir/assets/methods_description_template.yml", checkIfExists: true) +ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath( params.multiqc_config ) : Channel.empty() +ch_multiqc_logo = params.multiqc_logo ? Channel.fromPath( params.multiqc_logo ) : Channel.empty() +ch_multiqc_custom_methods_description = params.multiqc_methods_description ? file(params.multiqc_methods_description) : file("$projectDir/assets/methods_description_template.yml", checkIfExists: true) /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -34,7 +34,6 @@ ch_multiqc_custom_methods_description = params.multiqc_methods_description ? fil // // SUBWORKFLOW: Consisting of a mix of local and nf-core/modules // -include { INPUT_CHECK } from '../subworkflows/local/input_check' // // MODULE @@ -138,36 +137,43 @@ workflow CRISPRSEQ_TARGETED { ch_versions = Channel.empty() // - // SUBWORKFLOW: Read in samplesheet, validate and stage input files + // Create input channel from input file provided through params.input // - INPUT_CHECK ( - file(params.input) - ) + Channel.fromSamplesheet("input") + .multiMap { + meta, fastq_1, fastq_2, condition, reference, protospacer, template -> + reads: [ meta.id, meta + [ single_end:false, self_reference:reference?false:true, template:template?true:false ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] + reference: [meta + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], reference] + protospacer: [meta + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], protospacer] + template: [meta + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], template] + } + .set { ch_input } + + ch_input .reads + .groupTuple() .map { - meta, fastq -> - [ meta - meta.subMap('id') + [id: meta.id.split('_')[0..-2].join('_')], fastq ] + WorkflowCrisprseq.validateInput(it) } - .groupTuple(by: [0]) // Separate samples by the ones containing all reads in one file or the ones with many files to be concatenated .branch { - meta, fastq -> - single : fastq.size() == 1 - return [ meta, fastq.flatten() ] - multiple: fastq.size() > 1 - return [ meta, fastq.flatten() ] + meta, fastqs -> + single : fastqs.size() == 1 + return [ meta, fastqs.flatten() ] + multiple: fastqs.size() > 1 + return [ meta, fastqs.flatten() ] } .set { ch_fastq } - ch_versions = ch_versions.mix(INPUT_CHECK.out.versions) + // // MODULE: Add reference sequences to file // SEQ_TO_FILE_REF ( - INPUT_CHECK.out.reference + ch_input.reference .map { meta, fastq -> - [ meta - meta.subMap('id') + [id: meta.id.split('_')[0..-2].join('_')], fastq ] + [ meta , fastq ] }, "reference" ) @@ -177,10 +183,10 @@ workflow CRISPRSEQ_TARGETED { // MODULE: Add template sequences to file // SEQ_TO_FILE_TEMPL ( - INPUT_CHECK.out.template + ch_input.template .map { meta, fastq -> - [ meta - meta.subMap('id') + [id: meta.id.split('_')[0..-2].join('_')], fastq ] + [ meta , fastq ] }, "template" ) @@ -190,10 +196,10 @@ workflow CRISPRSEQ_TARGETED { // to channel: [ meta, reference, protospacer] if (!params.reference_fasta && !params.protospacer) { SEQ_TO_FILE_REF.out.file - .join(INPUT_CHECK.out.protospacer + .join(ch_input.protospacer .map { meta, fastq -> - [ meta - meta.subMap('id') + [id: meta.id.split('_')[0..-2].join('_')], fastq ] + [ meta , fastq ] }, by: 0) .set{ reference_protospacer } @@ -206,20 +212,20 @@ workflow CRISPRSEQ_TARGETED { } else if (!params.protospacer) { // If a reference was provided through a fasta file or igenomes instead of the samplesheet ch_reference = Channel.fromPath(params.reference_fasta) - INPUT_CHECK.out.protospacer + ch_input.protospacer .combine(ch_reference) .map{ meta, protospacer, reference -> - [ meta - meta.subMap('id') + [id: meta.id.split('_')[0..-2].join('_')], reference, protospacer ] + [ meta , reference, protospacer ] } .set{ reference_protospacer } } else { ch_reference = Channel.fromPath(params.reference_fasta) ch_protospacer = Channel.of(params.protospacer) - INPUT_CHECK.out.reads + ch_input.reads .combine(ch_reference) .combine(ch_protospacer) .map{ meta, reads, reference, protospacer -> - [meta - meta.subMap('id') + [id: meta.id.split('_')[0..-2].join('_')], reference, protospacer] + [meta , reference, protospacer] } .set{ reference_protospacer } } @@ -681,10 +687,10 @@ workflow CRISPRSEQ_TARGETED { ch_mapped_bam .join(SAMTOOLS_INDEX.out.bai) .join(ORIENT_REFERENCE.out.reference) - .join(INPUT_CHECK.out.protospacer + .join(ch_input.protospacer .map { meta, fastq -> - [ meta - meta.subMap('id') + [id: meta.id.split('_')[0..-2].join('_')], fastq ] + [ meta , fastq ] } ) .join(SEQ_TO_FILE_TEMPL.out.file, remainder: true) From cbfd31683285e3ec20ef07946e3ca47d3e7be3d8 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 18 Jul 2023 15:30:30 +0200 Subject: [PATCH 014/133] delete input check scripts --- bin/check_samplesheet.py | 259 ----------------------------- modules/local/samplesheet_check.nf | 31 ---- subworkflows/local/input_check.nf | 130 --------------- workflows/crisprseq_targeted.nf | 2 +- 4 files changed, 1 insertion(+), 421 deletions(-) delete mode 100755 bin/check_samplesheet.py delete mode 100644 modules/local/samplesheet_check.nf delete mode 100644 subworkflows/local/input_check.nf diff --git a/bin/check_samplesheet.py b/bin/check_samplesheet.py deleted file mode 100755 index 4a758fe0..00000000 --- a/bin/check_samplesheet.py +++ /dev/null @@ -1,259 +0,0 @@ -#!/usr/bin/env python - - -"""Provide a command line tool to validate and transform tabular samplesheets.""" - - -import argparse -import csv -import logging -import sys -from collections import Counter -from pathlib import Path - -logger = logging.getLogger() - - -class RowChecker: - """ - Define a service that can validate and transform each given row. - - Attributes: - modified (list): A list of dicts, where each dict corresponds to a previously - validated and transformed row. The order of rows is maintained. - - """ - - VALID_FORMATS = ( - ".fq.gz", - ".fastq.gz", - ) - - def __init__( - self, - sample_col="sample", - first_col="fastq_1", - second_col="fastq_2", - single_col="single_end", - **kwargs, - ): - """ - Initialize the row checker with the expected column names. - - Args: - sample_col (str): The name of the column that contains the sample name - (default "sample"). - first_col (str): The name of the column that contains the first (or only) - FASTQ file path (default "fastq_1"). - second_col (str): The name of the column that contains the second (if any) - FASTQ file path (default "fastq_2"). - single_col (str): The name of the new column that will be inserted and - records whether the sample contains single- or paired-end sequencing - reads (default "single_end"). - - """ - super().__init__(**kwargs) - self._sample_col = sample_col - self._first_col = first_col - self._second_col = second_col - self._single_col = single_col - self._seen = set() - self.modified = [] - - def validate_and_transform(self, row): - """ - Perform all validations on the given row and insert the read pairing status. - - Args: - row (dict): A mapping from column headers (keys) to elements of that row - (values). - - """ - self._validate_sample(row) - self._validate_first(row) - self._validate_second(row) - self._validate_pair(row) - self._seen.add((row[self._sample_col], row[self._first_col])) - self.modified.append(row) - - def _validate_sample(self, row): - """Assert that the sample name exists and convert spaces to underscores.""" - if len(row[self._sample_col]) <= 0: - raise AssertionError("Sample input is required.") - # Sanitize samples slightly. - row[self._sample_col] = row[self._sample_col].replace(" ", "_") - - def _validate_first(self, row): - """Assert that the first FASTQ entry is non-empty and has the right format.""" - if len(row[self._first_col]) <= 0: - raise AssertionError("At least the first FASTQ file is required.") - self._validate_fastq_format(row[self._first_col]) - - def _validate_second(self, row): - """Assert that the second FASTQ entry has the right format if it exists.""" - if len(row[self._second_col]) > 0: - self._validate_fastq_format(row[self._second_col]) - - def _validate_pair(self, row): - """Assert that read pairs have the same file extension. Report pair status.""" - if row[self._first_col] and row[self._second_col]: - row[self._single_col] = False - first_col_suffix = Path(row[self._first_col]).suffixes[-2:] - second_col_suffix = Path(row[self._second_col]).suffixes[-2:] - if first_col_suffix != second_col_suffix: - raise AssertionError("FASTQ pairs must have the same file extensions.") - else: - row[self._single_col] = True - - def _validate_fastq_format(self, filename): - """Assert that a given filename has one of the expected FASTQ extensions.""" - if not any(filename.endswith(extension) for extension in self.VALID_FORMATS): - raise AssertionError( - f"The FASTQ file has an unrecognized extension: {filename}\n" - f"It should be one of: {', '.join(self.VALID_FORMATS)}" - ) - - def validate_unique_samples(self): - """ - Assert that the combination of sample name and FASTQ filename is unique. - - In addition to the validation, also rename all samples to have a suffix of _T{n}, where n is the - number of times the same sample exist, but with different FASTQ files, e.g., multiple runs per experiment. - - """ - if len(self._seen) != len(self.modified): - raise AssertionError("The pair of sample name and FASTQ must be unique.") - seen = Counter() - for row in self.modified: - sample = row[self._sample_col] - seen[sample] += 1 - row[self._sample_col] = f"{sample}_T{seen[sample]}" - - -def read_head(handle, num_lines=10): - """Read the specified number of lines from the current position in the file.""" - lines = [] - for idx, line in enumerate(handle): - if idx == num_lines: - break - lines.append(line) - return "".join(lines) - - -def sniff_format(handle): - """ - Detect the tabular format. - - Args: - handle (text file): A handle to a `text file`_ object. The read position is - expected to be at the beginning (index 0). - - Returns: - csv.Dialect: The detected tabular format. - - .. _text file: - https://docs.python.org/3/glossary.html#term-text-file - - """ - peek = read_head(handle) - handle.seek(0) - sniffer = csv.Sniffer() - dialect = sniffer.sniff(peek) - return dialect - - -def check_samplesheet(file_in, file_out): - """ - Check that the tabular samplesheet has the structure expected by nf-core pipelines. - - Validate the general shape of the table, expected columns, and each row. Also add - an additional column which records whether one or two FASTQ reads were found. - - Args: - file_in (pathlib.Path): The given tabular samplesheet. The format can be either - CSV, TSV, or any other format automatically recognized by ``csv.Sniffer``. - file_out (pathlib.Path): Where the validated and transformed samplesheet should - be created; always in CSV format. - - Example: - This function checks that the samplesheet follows the following structure, - see also the `viral recon samplesheet`_:: - - sample,fastq_1,fastq_2 - SAMPLE_PE,SAMPLE_PE_RUN1_1.fastq.gz,SAMPLE_PE_RUN1_2.fastq.gz - SAMPLE_PE,SAMPLE_PE_RUN2_1.fastq.gz,SAMPLE_PE_RUN2_2.fastq.gz - SAMPLE_SE,SAMPLE_SE_RUN1_1.fastq.gz, - - .. _viral recon samplesheet: - https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/samplesheet/samplesheet_test_illumina_amplicon.csv - - """ - required_columns = {"sample", "fastq_1", "fastq_2"} - # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`. - with file_in.open(newline="") as in_handle: - reader = csv.DictReader(in_handle, dialect=sniff_format(in_handle)) - # Validate the existence of the expected header columns. - if not required_columns.issubset(reader.fieldnames): - req_cols = ", ".join(required_columns) - logger.critical(f"The sample sheet **must** contain these column headers: {req_cols}.") - sys.exit(1) - # Validate each row. - checker = RowChecker() - for i, row in enumerate(reader): - try: - checker.validate_and_transform(row) - except AssertionError as error: - logger.critical(f"{str(error)} On line {i + 2}.") - sys.exit(1) - checker.validate_unique_samples() - header = list(reader.fieldnames) - header.insert(1, "single_end") - # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`. - with file_out.open(mode="w", newline="") as out_handle: - writer = csv.DictWriter(out_handle, header, delimiter=",") - writer.writeheader() - for row in checker.modified: - writer.writerow(row) - - -def parse_args(argv=None): - """Define and immediately parse command line arguments.""" - parser = argparse.ArgumentParser( - description="Validate and transform a tabular samplesheet.", - epilog="Example: python check_samplesheet.py samplesheet.csv samplesheet.valid.csv", - ) - parser.add_argument( - "file_in", - metavar="FILE_IN", - type=Path, - help="Tabular input samplesheet in CSV or TSV format.", - ) - parser.add_argument( - "file_out", - metavar="FILE_OUT", - type=Path, - help="Transformed output samplesheet in CSV format.", - ) - parser.add_argument( - "-l", - "--log-level", - help="The desired log level (default WARNING).", - choices=("CRITICAL", "ERROR", "WARNING", "INFO", "DEBUG"), - default="WARNING", - ) - return parser.parse_args(argv) - - -def main(argv=None): - """Coordinate argument parsing and program execution.""" - args = parse_args(argv) - logging.basicConfig(level=args.log_level, format="[%(levelname)s] %(message)s") - if not args.file_in.is_file(): - logger.error(f"The given input file {args.file_in} was not found!") - sys.exit(2) - args.file_out.parent.mkdir(parents=True, exist_ok=True) - check_samplesheet(args.file_in, args.file_out) - - -if __name__ == "__main__": - sys.exit(main()) diff --git a/modules/local/samplesheet_check.nf b/modules/local/samplesheet_check.nf deleted file mode 100644 index ec0e11b2..00000000 --- a/modules/local/samplesheet_check.nf +++ /dev/null @@ -1,31 +0,0 @@ -process SAMPLESHEET_CHECK { - tag "$samplesheet" - label 'process_single' - - conda "conda-forge::python=3.8.3" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/python:3.8.3' : - 'biocontainers/python:3.8.3' }" - - input: - path samplesheet - - output: - path '*.csv' , emit: csv - path "versions.yml", emit: versions - - when: - task.ext.when == null || task.ext.when - - script: // This script is bundled with the pipeline, in nf-core/crisprseq/bin/ - """ - check_samplesheet.py \\ - $samplesheet \\ - samplesheet.valid.csv - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - python: \$(python --version | sed 's/Python //g') - END_VERSIONS - """ -} diff --git a/subworkflows/local/input_check.nf b/subworkflows/local/input_check.nf deleted file mode 100644 index c9aafbde..00000000 --- a/subworkflows/local/input_check.nf +++ /dev/null @@ -1,130 +0,0 @@ -// -// Check input samplesheet and get read channels -// - -include { SAMPLESHEET_CHECK } from '../../modules/local/samplesheet_check' - -workflow INPUT_CHECK { - take: - samplesheet // file: /path/to/samplesheet.csv - - main: - SAMPLESHEET_CHECK ( samplesheet ) - .csv - .splitCsv ( header:true, sep:',' ) - .multiMap { - reads: create_fastq_channel(it) - reference: create_reference_channel(it) - template: create_template_channel(it) - protospacer: create_protospacer_channel(it) - } - .set { inputs } - - emit: - reads = inputs.reads // channel: [ val(meta), [ reads ] ] - reference = inputs.reference // channel: [ val(meta), reference ] - template = inputs.template // channel: [ val(meta), template ] - protospacer = inputs.protospacer // channel: [ val(meta), protospacer ] - versions = SAMPLESHEET_CHECK.out.versions // channel: [ versions.yml ] - -} - -// Function to generate meta map -def create_meta(LinkedHashMap row) { - // Create meta map with sample ID - def meta = [:] - meta.id = row.sample - - // Add single end boolean - if (row.fastq_2 && file(row.fastq_2).exists()) { - meta.single_end = false - } else { - meta.single_end = true - } - - // Add self reference boolean - if (row.reference.length() <= 0) { - meta.self_reference = true - } else { - meta.self_reference = false - } - - // Add template boolean - if (!row.template) { - meta.template = false - } else if (!file(row.template).exists()) { - meta.template = true - } else { - meta.template = true - } - - return meta -} - -// Function to get list of [ meta, [ fastq_1, fastq_2 ] ] -def create_fastq_channel(LinkedHashMap row) { - // create meta map - def meta = create_meta(row) - - // add path(s) of the fastq file(s) to the meta map - def fastq_meta = [] - if (!file(row.fastq_1).exists()) { - error("ERROR: Please check input samplesheet -> Read 1 FastQ file does not exist!\n${row.fastq_1}") - } - if (!row.fastq_2 || !file(row.fastq_2).exists()) { - fastq_meta = [ meta, [ file(row.fastq_1) ] ] - } else { - fastq_meta = [ meta, [ file(row.fastq_1), file(row.fastq_2) ] ] - } - return fastq_meta -} - -// Function to get a list of [ meta, reference ] -def create_reference_channel(LinkedHashMap row) { - // create meta map - def meta = create_meta(row) - - // add reference sequence to meta - def reference_meta = [] - if (row.reference.length() <= 0) { - reference_meta = [ meta, null ] - } else { - reference_meta = [ meta, row.reference ] - } - - return reference_meta -} - -// Function to get a list of [ meta, template ] -def create_template_channel(LinkedHashMap row) { - // create meta map - def meta = create_meta(row) - - // add template sequence/path to meta - def template_meta = [] - if (!row.template) { - template_meta = [ meta, null ] - } else if (!file(row.template).exists()) { - template_meta = [ meta, row.template ] - } else { - template_meta = [ meta, file(row.template) ] - } - - return template_meta -} - -// Function to get a list of [ meta, protospacer ] -def create_protospacer_channel(LinkedHashMap row) { - // create meta map - def meta = create_meta(row) - - // add protospacer sequence to meta - def protospacer_meta = [] - if (row.protospacer.length() <= 0 && !params.protospacer) { - error("ERROR: Please check input samplesheet -> Protospacer sequence is not provided!\n") - } else { - protospacer_meta = [ meta, row.protospacer ] - } - - return protospacer_meta -} diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 8f4d33f7..0229c4b9 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -141,7 +141,7 @@ workflow CRISPRSEQ_TARGETED { // Channel.fromSamplesheet("input") .multiMap { - meta, fastq_1, fastq_2, condition, reference, protospacer, template -> + meta, fastq_1, fastq_2, x, reference, protospacer, template -> reads: [ meta.id, meta + [ single_end:false, self_reference:reference?false:true, template:template?true:false ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] reference: [meta + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], reference] protospacer: [meta + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], protospacer] From 7e4a20569603a97be756adea2eea8a716afda6f4 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 18 Jul 2023 15:39:41 +0200 Subject: [PATCH 015/133] add fromSamplesheet to screening --- assets/schema_input.json | 3 +- bin/check_samplesheet_screening.py | 253 ------------------- modules/local/samplesheet_check_screening.nf | 28 -- subworkflows/local/input_check_screening.nf | 44 ---- workflows/crisprseq_screening.nf | 20 +- workflows/crisprseq_targeted.nf | 12 +- 6 files changed, 21 insertions(+), 339 deletions(-) delete mode 100755 bin/check_samplesheet_screening.py delete mode 100644 modules/local/samplesheet_check_screening.nf delete mode 100644 subworkflows/local/input_check_screening.nf diff --git a/assets/schema_input.json b/assets/schema_input.json index a66228ed..ea6d0647 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -38,7 +38,8 @@ "type": "string", "pattern": "^\\S+$", "enum": ["control", "treatment"], - "errorMessage": "Condition name must be one of 'control' or 'treatment'" + "errorMessage": "Condition name must be one of 'control' or 'treatment'", + "meta": ["condition"] }, "reference": { "type": "string", diff --git a/bin/check_samplesheet_screening.py b/bin/check_samplesheet_screening.py deleted file mode 100755 index c069109d..00000000 --- a/bin/check_samplesheet_screening.py +++ /dev/null @@ -1,253 +0,0 @@ -#!/usr/bin/env python - - -"""Provide a command line tool to validate and transform tabular samplesheets.""" - - -import argparse -import csv -import logging -import sys -from collections import Counter -from pathlib import Path - -logger = logging.getLogger() - - -class RowChecker: - """ - Define a service that can validate and transform each given row. - - Attributes: - modified (list): A list of dicts, where each dict corresponds to a previously - validated and transformed row. The order of rows is maintained. - - """ - - VALID_FORMATS = ( - ".fq.gz", - ".fastq.gz", - ) - - def __init__( - self, - sample_col="sample", - first_col="fastq_1", - second_col="fastq_2", - condition="control", - **kwargs, - ): - """ - Initialize the row checker with the expected column names. - - Args: - sample_col (str): The name of the column that contains the sample name - (default "sample"). - first_col (str): The name of the column that contains the first (or only) - FASTQ file path (default "fastq_1"). - second_col (str): The name of the column that contains the second (if any) - FASTQ file path (default "fastq_2"). - condition (str): The name of the condition of the sample. - - """ - super().__init__(**kwargs) - self._sample_col = sample_col - self._first_col = first_col - self._second_col = second_col - self._condition = condition - self._seen = set() - self.modified = [] - - def validate_and_transform(self, row): - """ - Perform all validations on the given row and insert the read pairing status. - - Args: - row (dict): A mapping from column headers (keys) to elements of that row - (values). - - """ - self._validate_sample(row) - self._validate_first(row) - self._validate_second(row) - self._validate_pair(row) - self._seen.add((row[self._sample_col], row[self._first_col])) - self.modified.append(row) - - def _validate_sample(self, row): - """Assert that the sample name exists and convert spaces to underscores.""" - if len(row[self._sample_col]) <= 0: - raise AssertionError("Sample input is required.") - # Sanitize samples slightly. - row[self._sample_col] = row[self._sample_col].replace(" ", "_") - - def _validate_first(self, row): - """Assert that the first FASTQ entry is non-empty and has the right format.""" - if len(row[self._first_col]) <= 0: - raise AssertionError("At least the first FASTQ file is required.") - self._validate_fastq_format(row[self._first_col]) - - def _validate_second(self, row): - """Assert that the second FASTQ entry has the right format if it exists.""" - if len(row[self._second_col]) > 0: - self._validate_fastq_format(row[self._second_col]) - - def _validate_pair(self, row): - """Assert that read pairs have the same file extension. Report pair status.""" - if row[self._first_col] and row[self._second_col]: - first_col_suffix = Path(row[self._first_col]).suffixes[-2:] - second_col_suffix = Path(row[self._second_col]).suffixes[-2:] - if first_col_suffix != second_col_suffix: - raise AssertionError("FASTQ pairs must have the same file extensions.") - - def _validate_fastq_format(self, filename): - """Assert that a given filename has one of the expected FASTQ extensions.""" - if not any(filename.endswith(extension) for extension in self.VALID_FORMATS): - raise AssertionError( - f"The FASTQ file has an unrecognized extension: {filename}\n" - f"It should be one of: {', '.join(self.VALID_FORMATS)}" - ) - - def validate_unique_samples(self): - """ - Assert that the combination of sample name and FASTQ filename is unique. - - In addition to the validation, also rename all samples to have a suffix of _T{n}, where n is the - number of times the same sample exist, but with different FASTQ files, e.g., multiple runs per experiment. - - """ - if len(self._seen) != len(self.modified): - raise AssertionError("The pair of sample name and FASTQ must be unique.") - seen = Counter() - for row in self.modified: - sample = row[self._sample_col] - seen[sample] += 1 - row[self._sample_col] = f"{sample}_T{seen[sample]}" - - -def read_head(handle, num_lines=10): - """Read the specified number of lines from the current position in the file.""" - lines = [] - for idx, line in enumerate(handle): - if idx == num_lines: - break - lines.append(line) - return "".join(lines) - - -def sniff_format(handle): - """ - Detect the tabular format. - - Args: - handle (text file): A handle to a `text file`_ object. The read position is - expected to be at the beginning (index 0). - - Returns: - csv.Dialect: The detected tabular format. - - .. _text file: - https://docs.python.org/3/glossary.html#term-text-file - - """ - peek = read_head(handle) - handle.seek(0) - sniffer = csv.Sniffer() - dialect = sniffer.sniff(peek) - return dialect - - -def check_samplesheet(file_in, file_out): - """ - Check that the tabular samplesheet has the structure expected by nf-core pipelines. - - Validate the general shape of the table, expected columns, and each row. Also add - an additional column which records whether one or two FASTQ reads were found. - - Args: - file_in (pathlib.Path): The given tabular samplesheet. The format can be either - CSV, TSV, or any other format automatically recognized by ``csv.Sniffer``. - file_out (pathlib.Path): Where the validated and transformed samplesheet should - be created; always in CSV format. - - Example: - This function checks that the samplesheet follows the following structure, - see also the `viral recon samplesheet`_:: - - sample,fastq_1,fastq_2,condition - SAMPLE_PE,SAMPLE_PE_RUN1_1.fastq.gz,SAMPLE_PE_RUN1_2.fastq.gz,control - SAMPLE_SE,SAMPLE_SE_RUN1_1.fastq.gz,,tratment - - .. _viral recon samplesheet: - https://raw.githubusercontent.com/nf-core/test-datasets/viralrecon/samplesheet/samplesheet_test_illumina_amplicon.csv - - """ - required_columns = {"sample", "fastq_1", "fastq_2"} - # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`. - with file_in.open(newline="") as in_handle: - reader = csv.DictReader(in_handle, dialect=sniff_format(in_handle)) - # Validate the existence of the expected header columns. - if not required_columns.issubset(reader.fieldnames): - req_cols = ", ".join(required_columns) - logger.critical(f"The sample sheet **must** contain these column headers: {req_cols}.") - sys.exit(1) - # Validate each row. - checker = RowChecker() - for i, row in enumerate(reader): - try: - checker.validate_and_transform(row) - except AssertionError as error: - logger.critical(f"{str(error)} On line {i + 2}.") - sys.exit(1) - checker.validate_unique_samples() - header = list(reader.fieldnames) - header.insert(1, "single_end") - # See https://docs.python.org/3.9/library/csv.html#id3 to read up on `newline=""`. - with file_out.open(mode="w", newline="") as out_handle: - writer = csv.DictWriter(out_handle, header, delimiter=",") - writer.writeheader() - for row in checker.modified: - writer.writerow(row) - - -def parse_args(argv=None): - """Define and immediately parse command line arguments.""" - parser = argparse.ArgumentParser( - description="Validate and transform a tabular samplesheet.", - epilog="Example: python check_samplesheet.py samplesheet.csv samplesheet.valid.csv", - ) - parser.add_argument( - "file_in", - metavar="FILE_IN", - type=Path, - help="Tabular input samplesheet in CSV or TSV format.", - ) - parser.add_argument( - "file_out", - metavar="FILE_OUT", - type=Path, - help="Transformed output samplesheet in CSV format.", - ) - parser.add_argument( - "-l", - "--log-level", - help="The desired log level (default WARNING).", - choices=("CRITICAL", "ERROR", "WARNING", "INFO", "DEBUG"), - default="WARNING", - ) - return parser.parse_args(argv) - - -def main(argv=None): - """Coordinate argument parsing and program execution.""" - args = parse_args(argv) - logging.basicConfig(level=args.log_level, format="[%(levelname)s] %(message)s") - if not args.file_in.is_file(): - logger.error(f"The given input file {args.file_in} was not found!") - sys.exit(2) - args.file_out.parent.mkdir(parents=True, exist_ok=True) - check_samplesheet(args.file_in, args.file_out) - - -if __name__ == "__main__": - sys.exit(main()) diff --git a/modules/local/samplesheet_check_screening.nf b/modules/local/samplesheet_check_screening.nf deleted file mode 100644 index 69b887f6..00000000 --- a/modules/local/samplesheet_check_screening.nf +++ /dev/null @@ -1,28 +0,0 @@ -process SAMPLESHEET_CHECK_SCREENING { - tag "$samplesheet" - label 'process_single' - - conda "conda-forge::python=3.8.3" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/python:3.8.3' : - 'biocontainers/python:3.8.3' }" - - input: - path samplesheet - - output: - path '*.csv' , emit: csv - path "versions.yml", emit: versions - - script: // This script is bundled with the pipeline, in nf-core/test/bin/ - """ - check_samplesheet_screening.py \\ - $samplesheet \\ - samplesheet.valid.csv - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - python: \$(python --version | sed 's/Python //g') - END_VERSIONS - """ -} diff --git a/subworkflows/local/input_check_screening.nf b/subworkflows/local/input_check_screening.nf deleted file mode 100644 index 8b3ba471..00000000 --- a/subworkflows/local/input_check_screening.nf +++ /dev/null @@ -1,44 +0,0 @@ -// -// Check input samplesheet and get read channels -// - -include { SAMPLESHEET_CHECK_SCREENING } from '../../modules/local/samplesheet_check_screening' - -workflow INPUT_CHECK_SCREENING { - take: - samplesheet // file: /path/to/samplesheet.csv - - main: - SAMPLESHEET_CHECK_SCREENING ( samplesheet ) - .csv - .splitCsv ( header:true, sep:',' ) - .map { create_fastq_channel(it) } - .set { reads } - - emit: - reads // channel: [ val(meta), [ reads ] ] - versions = SAMPLESHEET_CHECK_SCREENING.out.versions // channel: [ versions.yml ] -} - -// Function to get list of [ meta, [ fastq_1, fastq_2 ] ] -def create_fastq_channel(LinkedHashMap row) { - // create meta map - def meta = [:] - meta.id = row.sample - meta.condition = row.condition - - // add path(s) of the fastq file(s) to the meta map - def fastq_meta = [] - if (!file(row.fastq_1).exists()) { - error("ERROR: Please check input samplesheet -> Read 1 FastQ file does not exist!\n${row.fastq_1}") - } - if (!row.fastq_2) { - fastq_meta = [ meta, [ file(row.fastq_1) ] ] - } else { - if (!file(row.fastq_2).exists()) { - error("ERROR: Please check input samplesheet -> Read 2 FastQ file does not exist!\n${row.fastq_2}") - } - fastq_meta = [ meta, [ file(row.fastq_1), file(row.fastq_2) ] ] - } - return fastq_meta -} diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index ff6fcada..fdff3857 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -76,22 +76,28 @@ workflow CRISPRSEQ_SCREENING { if(!params.count_table){ // - // SUBWORKFLOW: Read in samplesheet, validate and stage input files + // Create input channel from input file provided through params.input // - INPUT_CHECK_SCREENING ( - file(params.input) - ) - ch_versions = ch_versions.mix(INPUT_CHECK_SCREENING.out.versions) + Channel.fromSamplesheet("input") + .map{ meta, fastq_1, fastq_2, x, y, z -> + // x (reference), y (protospacer), and z (template) are part of the targeted workflows and we don't need them + if (!fastq_2) { + return [ meta, [ fastq_1 ] ] + } else { + return [ meta, [ fastq_1, fastq_2 ] ] + } + } + .set { ch_input } // // MODULE: Run FastQC // FASTQC ( - INPUT_CHECK_SCREENING.out.reads + ch_input ) ch_versions = ch_versions.mix(FASTQC.out.versions.first()) - INPUT_CHECK_SCREENING.out.reads + ch_input .map { meta, fastq -> [meta.condition, fastq] } diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 0229c4b9..49ab46c5 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -140,12 +140,12 @@ workflow CRISPRSEQ_TARGETED { // Create input channel from input file provided through params.input // Channel.fromSamplesheet("input") - .multiMap { - meta, fastq_1, fastq_2, x, reference, protospacer, template -> - reads: [ meta.id, meta + [ single_end:false, self_reference:reference?false:true, template:template?true:false ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] - reference: [meta + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], reference] - protospacer: [meta + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], protospacer] - template: [meta + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], template] + .multiMap { meta, fastq_1, fastq_2, reference, protospacer, template -> + // meta.condition is part of the screening pipeline and we need to remove it + reads: [ meta.id, meta - meta.subMap('condition') + [ single_end:false, self_reference:reference?false:true, template:template?true:false ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] + reference: [meta - meta.subMap('condition') + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], reference] + protospacer: [meta - meta.subMap('condition') + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], protospacer] + template: [meta - meta.subMap('condition') + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], template] } .set { ch_input } From 6cb5f6437663e3615606057f551e73ab32cc92d8 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 18 Jul 2023 15:42:32 +0200 Subject: [PATCH 016/133] update changelog --- CHANGELOG.md | 1 + workflows/crisprseq_targeted.nf | 2 +- 2 files changed, 2 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 02d741b5..90d0c5fd 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -8,6 +8,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Added - Template update v2.9 ([#52](https://github.com/nf-core/crisprseq/pull/52)) +- Use Channel.fromSamplesheet() from nf-validation to validate input sample sheets and create an input channel ([#54](https://github.com/nf-core/crisprseq/pull/54)) ### Fixed diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 49ab46c5..7aa25e77 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -141,7 +141,7 @@ workflow CRISPRSEQ_TARGETED { // Channel.fromSamplesheet("input") .multiMap { meta, fastq_1, fastq_2, reference, protospacer, template -> - // meta.condition is part of the screening pipeline and we need to remove it + // meta.condition is part of the screening workflow and we need to remove it reads: [ meta.id, meta - meta.subMap('condition') + [ single_end:false, self_reference:reference?false:true, template:template?true:false ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] reference: [meta - meta.subMap('condition') + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], reference] protospacer: [meta - meta.subMap('condition') + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], protospacer] From b24a00c60934d96aec99b189261b5ee2c8ff1ce9 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 18 Jul 2023 15:55:57 +0200 Subject: [PATCH 017/133] allow sequences in lower case --- CHANGELOG.md | 2 +- assets/schema_input.json | 6 +++--- 2 files changed, 4 insertions(+), 4 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 90d0c5fd..cc4ab20e 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -8,7 +8,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Added - Template update v2.9 ([#52](https://github.com/nf-core/crisprseq/pull/52)) -- Use Channel.fromSamplesheet() from nf-validation to validate input sample sheets and create an input channel ([#54](https://github.com/nf-core/crisprseq/pull/54)) +- Use `Channel.fromSamplesheet()` from `nf-validation` to validate input sample sheets and create an input channel ([#54](https://github.com/nf-core/crisprseq/pull/54)) ### Fixed diff --git a/assets/schema_input.json b/assets/schema_input.json index ea6d0647..13f50606 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -43,17 +43,17 @@ }, "reference": { "type": "string", - "pattern": "^[ACTGN]+$", + "pattern": "^[ACTGNactgn]+$", "errorMessage": "Reference sequence must be a valid DNA sequence" }, "protospacer": { "type": "string", - "pattern": "^[ACTGN]+$", + "pattern": "^[ACTGNactgn]+$", "errorMessage": "Protospacer sequence must be a valid DNA sequence" }, "template": { "type": "string", - "pattern": "^[ACTGN]+$", + "pattern": "^[ACTGNactgn]+$", "errorMessage": "Template sequence must be a valid DNA sequence" } }, From 808448ec50dfd7cc947ebce6fdda8ae5eb1f39c7 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Wed, 19 Jul 2023 14:25:06 +0200 Subject: [PATCH 018/133] replace seq_to_file module by channel operations --- modules/local/seq_to_file.nf | 33 --------------- workflows/crisprseq_targeted.nf | 75 ++++++++++++++++++--------------- 2 files changed, 41 insertions(+), 67 deletions(-) delete mode 100644 modules/local/seq_to_file.nf diff --git a/modules/local/seq_to_file.nf b/modules/local/seq_to_file.nf deleted file mode 100644 index 6dfc0022..00000000 --- a/modules/local/seq_to_file.nf +++ /dev/null @@ -1,33 +0,0 @@ -process SEQ_TO_FILE { - tag "$meta.id" - label 'process_single' - - conda "conda-forge::p7zip==16.02" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/p7zip:16.02' : - 'biocontainers/p7zip:16.02' }" - - input: - tuple val(meta), val(sequence) - val(type) - - output: - tuple val(meta), path('*.fasta') , emit: file - path "versions.yml" , emit: versions - - when: - sequence != null - - script: - def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" - """ - echo ">${meta.id}" > ${meta.id}_${type}.fasta - echo $sequence >> ${meta.id}_${type}.fasta - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - 7za: \$(echo \$(7za --help) | sed 's/.*p7zip Version //; s/(.*//') - END_VERSIONS - """ -} diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 7aa25e77..5b1f5a48 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -167,46 +167,59 @@ workflow CRISPRSEQ_TARGETED { // - // MODULE: Add reference sequences to file + // Add reference sequences to file // - SEQ_TO_FILE_REF ( - ch_input.reference - .map { - meta, fastq -> - [ meta , fastq ] - }, - "reference" + ch_input.reference + .tap{ meta_reference } + .collectFile() { meta, reference -> + [ "${meta.id}_reference.fasta", ">${meta.id}\n${item}\n" ] // Write each reference sequence to a file + } + .map{ new_file -> + [new_file.baseName.split("_reference")[0], new_file] // create a channel with the meta.id and the new file + } + .join(meta_reference + .map{ meta, reference -> + [meta.id, meta] // Join the channel by meta.id with the meta map + } ) - ch_versions = ch_versions.mix(SEQ_TO_FILE_REF.out.versions) + .map{ metaid, new_file, meta -> + [meta, new_file] // Obtain the final channel with meta map and the new file + } + .set{ ch_seq_reference } + // - // MODULE: Add template sequences to file + // Add template sequences to file // - SEQ_TO_FILE_TEMPL ( - ch_input.template - .map { - meta, fastq -> - [ meta , fastq ] - }, - "template" + ch_input.template + .tap{ meta_template } + .collectFile() { meta, template -> + [ "${meta.if}_template.fasta", ">${meta.id}\n${item}\n" ] // Write each template sequence to a file + } + .map{ new_file -> + [new_file.baseName.split("_template")[0], new_file] // create a channel with the meta.id and the new file + } + .join(meta_template + .map{ meta, template -> + [meta.id, meta] // Join the channel by meta.id with the meta map + } ) - ch_versions = ch_versions.mix(SEQ_TO_FILE_TEMPL.out.versions) + .map{ metaid, new_file, meta -> + [meta, new_file] // Obtain the final channel with meta map and the new file + } + .set{ ch_seq_template } + // Join channels with reference and protospacer // to channel: [ meta, reference, protospacer] if (!params.reference_fasta && !params.protospacer) { - SEQ_TO_FILE_REF.out.file - .join(ch_input.protospacer - .map { - meta, fastq -> - [ meta , fastq ] - }, - by: 0) + ch_seq_reference + .join(ch_input.protospacer) .set{ reference_protospacer } } else if (!params.reference_fasta) { // If a protospacer was provided through the --protospacer param instead of the samplesheet ch_protospacer = Channel.of(params.protospacer) - SEQ_TO_FILE_REF.out.file + ch_seq_reference .combine(ch_protospacer) .set{ reference_protospacer } } else if (!params.protospacer) { @@ -214,9 +227,6 @@ workflow CRISPRSEQ_TARGETED { ch_reference = Channel.fromPath(params.reference_fasta) ch_input.protospacer .combine(ch_reference) - .map{ meta, protospacer, reference -> - [ meta , reference, protospacer ] - } .set{ reference_protospacer } } else { ch_reference = Channel.fromPath(params.reference_fasta) @@ -224,9 +234,6 @@ workflow CRISPRSEQ_TARGETED { ch_input.reads .combine(ch_reference) .combine(ch_protospacer) - .map{ meta, reads, reference, protospacer -> - [meta , reference, protospacer] - } .set{ reference_protospacer } } @@ -656,7 +663,7 @@ workflow CRISPRSEQ_TARGETED { // TEMPLATE_REFERENCE ( ORIENT_REFERENCE.out.reference - .join(SEQ_TO_FILE_TEMPL.out.file) + .join(ch_seq_template) ) @@ -693,7 +700,7 @@ workflow CRISPRSEQ_TARGETED { [ meta , fastq ] } ) - .join(SEQ_TO_FILE_TEMPL.out.file, remainder: true) + .join(ch_seq_template, remainder: true) .join(ch_template_bam, remainder: true) .join(TEMPLATE_REFERENCE.out.fasta, remainder: true) .join(ALIGNMENT_SUMMARY.out.summary) From b053ff888421369ded41b73add68d9b6837908e5 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Wed, 19 Jul 2023 14:27:56 +0200 Subject: [PATCH 019/133] remove outdated module includes and comments --- workflows/crisprseq_targeted.nf | 8 +++----- 1 file changed, 3 insertions(+), 5 deletions(-) diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 5b1f5a48..56f1dfdf 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -40,8 +40,6 @@ ch_multiqc_custom_methods_description = params.multiqc_methods_description ? fil // include { FIND_ADAPTERS } from '../modules/local/find_adapters' include { EXTRACT_UMIS } from '../modules/local/extract_umis' -include { SEQ_TO_FILE as SEQ_TO_FILE_REF } from '../modules/local/seq_to_file' -include { SEQ_TO_FILE as SEQ_TO_FILE_TEMPL } from '../modules/local/seq_to_file' include { ORIENT_REFERENCE } from '../modules/local/orient_reference' include { CIGAR_PARSER } from '../modules/local/cigar_parser' include { MERGING_SUMMARY } from '../modules/local/merging_summary' @@ -447,11 +445,11 @@ workflow CRISPRSEQ_TARGETED { [ "${name}.fasta", fasta ] // >centroid_... -> sample_top.fasta } .map{ new_file -> - [new_file.baseName, new_file] // Substring is removing "_top" added by VSEARCH_SORT // [sample, sample_top.fasta] + [new_file.baseName, new_file] // [sample, sample_top.fasta] } .join(meta_channel_2 .map { meta, original_file -> - ["${original_file.baseName}", meta] // Substring is removing "_top" added by VSEARCH_SORT // [sample, [id:sample_id, ...]] + ["${original_file.baseName}", meta] // [sample, [id:sample_id, ...]] }) // [sample, sample_top.fasta, [id:sample_id, ...]] .map{ file_name, new_file, meta -> [meta + [cluster_id: file_name[0..-5]], new_file] // Add cluster ID to meta map // [[id:sample_id, ..., cluster_id:sample], sample_top.fasta] @@ -470,7 +468,7 @@ workflow CRISPRSEQ_TARGETED { [ "${name}_sequences.fasta", fasta ] // >... -> sample_sequences.fasta } .map{ new_file -> - [new_file.baseName[0..-11], new_file] // Substring is removing "_sequences" added by collectFile // [sample, sample_sequences.fasta] + [new_file.baseName[0..-11], new_file] // [sample, sample_sequences.fasta] } .join(meta_channel_3 .map { meta, original_file -> From f28c9fcd54397d0cad2cd1f68938c71fdab23f2b Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Wed, 19 Jul 2023 14:38:13 +0200 Subject: [PATCH 020/133] fix typos --- workflows/crisprseq_targeted.nf | 76 ++++++++++++++++----------------- 1 file changed, 38 insertions(+), 38 deletions(-) diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 56f1dfdf..02ad5f97 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -38,14 +38,14 @@ ch_multiqc_custom_methods_description = params.multiqc_methods_description ? fil // // MODULE // -include { FIND_ADAPTERS } from '../modules/local/find_adapters' -include { EXTRACT_UMIS } from '../modules/local/extract_umis' -include { ORIENT_REFERENCE } from '../modules/local/orient_reference' -include { CIGAR_PARSER } from '../modules/local/cigar_parser' -include { MERGING_SUMMARY } from '../modules/local/merging_summary' -include { CLUSTERING_SUMMARY } from '../modules/local/clustering_summary' -include { ALIGNMENT_SUMMARY } from '../modules/local/alignment_summary' -include { TEMPLATE_REFERENCE } from '../modules/local/template_reference' +include { FIND_ADAPTERS } from '../modules/local/find_adapters' +include { EXTRACT_UMIS } from '../modules/local/extract_umis' +include { ORIENT_REFERENCE } from '../modules/local/orient_reference' +include { CIGAR_PARSER } from '../modules/local/cigar_parser' +include { MERGING_SUMMARY } from '../modules/local/merging_summary' +include { CLUSTERING_SUMMARY } from '../modules/local/clustering_summary' +include { ALIGNMENT_SUMMARY } from '../modules/local/alignment_summary' +include { TEMPLATE_REFERENCE } from '../modules/local/template_reference' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -56,30 +56,30 @@ include { TEMPLATE_REFERENCE } from '../modules/loc // // MODULE: Installed directly from nf-core/modules // -include { FASTQC } from '../modules/nf-core/fastqc/main' -include { MULTIQC } from '../modules/nf-core/multiqc/main' -include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main' -include { PEAR } from '../modules/nf-core/pear/main' -include { CAT_FASTQ } from '../modules/nf-core/cat/fastq/main' -include { SEQTK_SEQ as SEQTK_SEQ_MASK } from '../modules/nf-core/seqtk/seq/main' -include { SEQTK_SEQ as SEQTK_SEQ_FATOFQ } from '../modules/nf-core/seqtk/seq/main' -include { VSEARCH_CLUSTER } from '../modules/nf-core/vsearch/cluster/main' -include { VSEARCH_SORT } from '../modules/nf-core/vsearch/sort/main' -include { RACON as RACON_1 } from '../modules/nf-core/racon/main' -include { RACON as RACON_2 } from '../modules/nf-core/racon/main' -include { BOWTIE2_ALIGN } from '../modules/nf-core/bowtie2/align/main' -include { BOWTIE2_BUILD } from '../modules/nf-core/bowtie2/build/main' -include { BWA_MEM } from '../modules/nf-core/bwa/mem/main' -include { BWA_INDEX } from '../modules/nf-core/bwa/index/main' -include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_ORIGINAL } from '../modules/nf-core/minimap2/align/main' -include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_UMI_1 } from '../modules/nf-core/minimap2/align/main' -include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_UMI_2 } from '../modules/nf-core/minimap2/align/main' -include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_TEMPLATE } from '../modules/nf-core/minimap2/align/main' -include { SAMTOOLS_FAIDX } from '../modules/nf-core/samtools/faidx/main' -include { MINIMAP2_INDEX } from '../modules/nf-core/minimap2/index/main' -include { MEDAKA } from '../modules/nf-core/medaka/main' -include { CUTADAPT } from '../modules/nf-core/cutadapt/main' -include { SAMTOOLS_INDEX } from '../modules/nf-core/samtools/index/main' +include { FASTQC } from '../modules/nf-core/fastqc/main' +include { MULTIQC } from '../modules/nf-core/multiqc/main' +include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main' +include { PEAR } from '../modules/nf-core/pear/main' +include { CAT_FASTQ } from '../modules/nf-core/cat/fastq/main' +include { SEQTK_SEQ as SEQTK_SEQ_MASK } from '../modules/nf-core/seqtk/seq/main' +include { SEQTK_SEQ as SEQTK_SEQ_FATOFQ } from '../modules/nf-core/seqtk/seq/main' +include { VSEARCH_CLUSTER } from '../modules/nf-core/vsearch/cluster/main' +include { VSEARCH_SORT } from '../modules/nf-core/vsearch/sort/main' +include { RACON as RACON_1 } from '../modules/nf-core/racon/main' +include { RACON as RACON_2 } from '../modules/nf-core/racon/main' +include { BOWTIE2_ALIGN } from '../modules/nf-core/bowtie2/align/main' +include { BOWTIE2_BUILD } from '../modules/nf-core/bowtie2/build/main' +include { BWA_MEM } from '../modules/nf-core/bwa/mem/main' +include { BWA_INDEX } from '../modules/nf-core/bwa/index/main' +include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_ORIGINAL } from '../modules/nf-core/minimap2/align/main' +include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_UMI_1 } from '../modules/nf-core/minimap2/align/main' +include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_UMI_2 } from '../modules/nf-core/minimap2/align/main' +include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_TEMPLATE } from '../modules/nf-core/minimap2/align/main' +include { SAMTOOLS_FAIDX } from '../modules/nf-core/samtools/faidx/main' +include { MINIMAP2_INDEX } from '../modules/nf-core/minimap2/index/main' +include { MEDAKA } from '../modules/nf-core/medaka/main' +include { CUTADAPT } from '../modules/nf-core/cutadapt/main' +include { SAMTOOLS_INDEX } from '../modules/nf-core/samtools/index/main' /* @@ -170,15 +170,15 @@ workflow CRISPRSEQ_TARGETED { ch_input.reference .tap{ meta_reference } .collectFile() { meta, reference -> - [ "${meta.id}_reference.fasta", ">${meta.id}\n${item}\n" ] // Write each reference sequence to a file + [ "${meta.id}_reference.fasta", ">${meta.id}\n${reference}\n" ] // Write each reference sequence to a file } .map{ new_file -> [new_file.baseName.split("_reference")[0], new_file] // create a channel with the meta.id and the new file } .join(meta_reference .map{ meta, reference -> - [meta.id, meta] // Join the channel by meta.id with the meta map - } + [meta.id, meta] // Join the channel by meta.id with the meta map + } ) .map{ metaid, new_file, meta -> [meta, new_file] // Obtain the final channel with meta map and the new file @@ -192,15 +192,15 @@ workflow CRISPRSEQ_TARGETED { ch_input.template .tap{ meta_template } .collectFile() { meta, template -> - [ "${meta.if}_template.fasta", ">${meta.id}\n${item}\n" ] // Write each template sequence to a file + [ "${meta.if}_template.fasta", ">${meta.id}\n${template}\n" ] // Write each template sequence to a file } .map{ new_file -> [new_file.baseName.split("_template")[0], new_file] // create a channel with the meta.id and the new file } .join(meta_template .map{ meta, template -> - [meta.id, meta] // Join the channel by meta.id with the meta map - } + [meta.id, meta] // Join the channel by meta.id with the meta map + } ) .map{ metaid, new_file, meta -> [meta, new_file] // Obtain the final channel with meta map and the new file From 237e7b8be4261034cb381c2308111d0f6655535d Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Wed, 19 Jul 2023 15:13:15 +0200 Subject: [PATCH 021/133] add suggestions from code review --- assets/schema_input.json | 9 +-------- workflows/crisprseq_targeted.nf | 8 ++++---- 2 files changed, 5 insertions(+), 12 deletions(-) diff --git a/assets/schema_input.json b/assets/schema_input.json index 13f50606..21c8c38a 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -25,14 +25,7 @@ "format": "file-path", "exists": true, "errorMessage": "FastQ file for reads 2 cannot contain spaces and must have extension '.fq.gz' or '.fastq.gz'", - "anyOf": [ - { - "pattern": "^\\S+\\.f(ast)?q\\.gz$" - }, - { - "maxLength": 0 - } - ] + "pattern": "^\\S+\\.f(ast)?q\\.gz$" }, "condition": { "type": "string", diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 02ad5f97..2e05a8f3 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -140,10 +140,10 @@ workflow CRISPRSEQ_TARGETED { Channel.fromSamplesheet("input") .multiMap { meta, fastq_1, fastq_2, reference, protospacer, template -> // meta.condition is part of the screening workflow and we need to remove it - reads: [ meta.id, meta - meta.subMap('condition') + [ single_end:false, self_reference:reference?false:true, template:template?true:false ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] - reference: [meta - meta.subMap('condition') + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], reference] - protospacer: [meta - meta.subMap('condition') + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], protospacer] - template: [meta - meta.subMap('condition') + [ single_end:true, self_reference:reference?false:true, template:template?true:false ], template] + reads: [ meta.id, meta - meta.subMap('condition') + [ single_end:fastq_2?false:true, self_reference:reference?false:true, template:template?true:false ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] + reference: [meta - meta.subMap('condition') + [ single_end:fastq_2?false:true, self_reference:reference?false:true, template:template?true:false ], reference] + protospacer: [meta - meta.subMap('condition') + [ single_end:fastq_2?false:true, self_reference:reference?false:true, template:template?true:false ], protospacer] + template: [meta - meta.subMap('condition') + [ single_end:fastq_2?false:true, self_reference:reference?false:true, template:template?true:false ], template] } .set { ch_input } From 3a506e65c1c3afd9f0c60e1cb42755d7b22c9c1f Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C3=BAlia=20Mir=20Pedrol?= Date: Wed, 19 Jul 2023 14:53:08 +0000 Subject: [PATCH 022/133] fix typo and filter out empty sequences --- conf/modules.config | 16 ---------------- workflows/crisprseq_targeted.nf | 4 +++- 2 files changed, 3 insertions(+), 17 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 43cdc492..75765bcd 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -18,22 +18,6 @@ process { saveAs: { filename -> filename.equals('versions.yml') ? null : filename } ] - withName: SEQ_TO_FILE_REF { - publishDir = [ - path: { "${params.outdir}/preprocessing/sequences" }, - mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } - ] - } - - withName: SEQ_TO_FILE_TEMPL { - publishDir = [ - path: { "${params.outdir}/preprocessing/sequences" }, - mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } - ] - } - withName: ORIENT_REFERENCE { ext.prefix = { params.reference_fasta ? "${reference.baseName}" : "${meta.id}_reference" } publishDir = [ diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 2e05a8f3..53c28942 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -169,6 +169,7 @@ workflow CRISPRSEQ_TARGETED { // ch_input.reference .tap{ meta_reference } + .filter{ meta, sequence -> sequence instanceof String } .collectFile() { meta, reference -> [ "${meta.id}_reference.fasta", ">${meta.id}\n${reference}\n" ] // Write each reference sequence to a file } @@ -191,8 +192,9 @@ workflow CRISPRSEQ_TARGETED { // ch_input.template .tap{ meta_template } + .filter{ meta, sequence -> sequence instanceof String } .collectFile() { meta, template -> - [ "${meta.if}_template.fasta", ">${meta.id}\n${template}\n" ] // Write each template sequence to a file + [ "${meta.id}_template.fasta", ">${meta.id}\n${template}\n" ] // Write each template sequence to a file } .map{ new_file -> [new_file.baseName.split("_template")[0], new_file] // create a channel with the meta.id and the new file From a748381fe3364ac487aced42e8f1068510343b7f Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C3=BAlia=20Mir=20Pedrol?= Date: Thu, 20 Jul 2023 08:51:53 +0200 Subject: [PATCH 023/133] Update CHANGELOG.md --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index cc4ab20e..e5d26623 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -8,7 +8,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Added - Template update v2.9 ([#52](https://github.com/nf-core/crisprseq/pull/52)) -- Use `Channel.fromSamplesheet()` from `nf-validation` to validate input sample sheets and create an input channel ([#54](https://github.com/nf-core/crisprseq/pull/54)) +- Use `Channel.fromSamplesheet()` from `nf-validation` to validate input sample sheets and create an input channel ([#58](https://github.com/nf-core/crisprseq/pull/58)) ### Fixed From b66f7e55c5badd38c17ed6371445d2e0cce62e7e Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Fri, 21 Jul 2023 08:55:47 +0200 Subject: [PATCH 024/133] Add bagel fold change --- bin/BAGEL.py | 1104 ++++++++++++++++++++++++++++++ modules/local/bagel2_fc.nf | 57 ++ workflows/crisprseq_screening.nf | 18 +- 3 files changed, 1177 insertions(+), 2 deletions(-) create mode 100755 bin/BAGEL.py create mode 100644 modules/local/bagel2_fc.nf diff --git a/bin/BAGEL.py b/bin/BAGEL.py new file mode 100755 index 00000000..6077e5dc --- /dev/null +++ b/bin/BAGEL.py @@ -0,0 +1,1104 @@ +#!/usr/bin/env python + +import click +import numpy as np +import pandas as pd +import scipy.stats as stats +from sklearn.linear_model import LinearRegression +import sys +import time + +VERSION = 2.0 + +BUILD = 115 + + + +''' +Update history + +Build 115 +1. Add single pass without resampling + +Build 114 +1. Add option for normalizing rep counts + +Build 113 +1. Fixed bugs + +Build 112 +1. Add sgRNA filtering options +2. Implemented 'Click' library. Thanks to John McGonigle + + +Build 111 +1. Add an option to equalize # of sgRNA per gene + +Build 110 +1. Enable multi control for fold-change calculation +2. Now user can input column names +3. Fix Threshold function + +''' + + +class OptionRequiredIf(click.Option): + + def full_process_value(self, ctx, value): + value = super(OptionRequiredIf, self).full_process_value(ctx, value) + + if value is None and ctx.params['filter_multi_target'] is True: + msg = 'Error! Multi-target-filtering selected and not align-info provided.\n' \ + 'Please indicate align-info file.' + raise click.MissingParameter(ctx=ctx, param=self, message=msg) + return value + + +# --------------------------------- +# BAGEL: Bayesian Analysis of Gene EssentaLity +# (c) Traver Hart , Eiru Kim 2017. + +# Acknowledgements: John McGonigle +# modified 10/2019 +# Free to modify and redistribute with attribution +# --------------------------------- + +# ------------------------------------ +# constants + + + +CONTEXT_SETTINGS = dict(help_option_names=['-h', '--help']) + + +def round_to_hundredth(x): + return np.around(x * 100) / 100.0 + + +def func_linear(x, a, b): + return (a * x) + b + + +class Training: + def __init__(self, X, n=None, cvnum=10): + + if n == None: + self._n = len(X) + self._cvnum = cvnum + self._bid = int(self._n / cvnum) + self._bucket = np.arange(len(X)) + self._X = X + self._step = 0 + + + def cross_validation(self): + if self._bid < 1: # bid check + print("The number of genes is too small! n<" + str(self._cvnum)) + sys.exit(1) + drawing = list() + mask = np.array([True] * self._n) + for j in range(self._bid): + # drawing.append(delete(self._bucket, np.random.randrange(len(self._bucket)))) + select = np.random.randint(len(self._bucket)) + drawing.append(self._bucket[select]) + mask[self._bucket[select]] = False + self._bucket = np.delete(self._bucket, select) + if self._step < self._n % self._cvnum: # for distribute remain.. + select = np.random.randint(len(self._bucket)) + drawing.append(self._bucket[select]) + mask[self._bucket[select]] = False + self._bucket = np.delete(self._bucket, select) + self._step += 1 + X_resample = self._X[mask] + return X_resample, self._X[~mask] + + def get_cv_step(self): + return self._step + + def bootstrap_resample(self): + mask = np.array([False] * self._n) + resample_i = np.floor(np.random.rand(self._n) * len(self._X)).astype(int) + + mask[resample_i] = True + X_resample = self._X[mask] + return X_resample, self._X[~mask] + + def get_data(self, method=0): + if method == 0: + train, test = self.bootstrap_resample() + elif method == 1: + train, test = self.cross_validation() + else: + print('Method passed as a value that was neither 0 nor 1.') + sys.exit(1) + return train, test + + +def fibo_weighted_sum(listofscore): + value = p1 = p2 = 0.0 + c = 1.0 # current value + for v in listofscore: + value += v / c + p2 = p1 # go one step + p1 = c + c = p1 + p2 + return value + + +# Note the \b in the doc string below is prevent click from wrapping the lines on the terminal. +@click.group(context_settings=CONTEXT_SETTINGS) +def bagel(): + """ + -------------------------------------------------------------------- + BAGEL.py + -------------------------------------------------------------------- + A tool from the Bayesian Analysis of Gene EssentiaLity (BAGEL) suite. + + \b + Calculate fold changes from read count data: + + \b + BAGEL.py fc -i [read count file] -o [output label] -c [control column] + + Calculate Bayes Factors from foldchange data: + + \b + BAGEL.py bf -i [fold change] -o [output file] -e [essentials genes] -n [nonessentials genes] -c [columns] + + + Calculate precision-recall from Bayes Factors: + + \b + BAGEL.py pr -i [Bayes Factor file] -o [output file] -e [essentials genes] -n [nonessentials genes] + + + + To print the current build and version use: + + \b + BAGEL.py version + """ + + +@click.command(name='version') +def report_bagel_version(): + """ + Report the current build and version no. + """ + print( + 'Bayesian Analysis of Gene EssentiaLity (BAGEL) suite:\n' + 'Version: {VERSION}\n' + 'Build: {BUILD}'.format(VERSION=VERSION, BUILD=BUILD) + ) + + +@click.command(name='fc') +@click.option('-i', '--read-count-file', required=True, type=click.Path(exists=True)) +@click.option('-o', '--output-label', required=True) +@click.option('-c', '--control-columns', required=True) +@click.option('-m', '--min-reads', type=int, default=0) +@click.option('-Np', '--pseudo-count', type=int, default=5) +def calculate_fold_change(read_count_file, output_label, control_columns, min_reads, pseudo_count): + """ + \b + Calculate fold changes from read count data outputting a fold change column: + + \b + BAGEL.py fc -i [read count file] -o [output label] -c [control column] + + \b + Required options: + -i --read-count-file Tab-delimited file of reagents and fold changes. See documentation for format. + -o --output-label Label for all output files + -c --control-columns A comma-delimited list of columns of control (T0 or plasmid) columns. + Input can be either number or name. + \b + Other options: + --min-reads=N Discard gRNA with T0 counts < N (default 0) + -Np, --pseudo-count=N Add a pseudocount of N to every readcount (default 5) + -h, --help Show this help text + + + \b + Example: + BAGEL.py fc -i readcount_file.txt -o experiment_name -c 1 + + This command calculates fold change, and writes [output label].foldchange and [output label].normalized_reads + + """ + + # ---------------------------------------------------------------- # + # Import raw read data, normalize, filter for T0 min read counts # + # Output: [output label].foldchange # + # ---------------------------------------------------------------- # + reads = pd.read_csv(read_count_file, sep='\t', index_col=0) + + reads[reads.columns.values[0]].fillna('NO_GENE_NAME', inplace=True) + reads.fillna(0, inplace=True) + control_columns = control_columns.split(",") + # + # check if controls are given as numeric or name + # + + try: + try: + ctrl_columns = list(map(int, control_columns)) + ctrl_labels = reads.columns.values[ctrl_columns] + except ValueError: + ctrl_labels = control_columns + + ctrl_sum = reads[ctrl_labels].sum(axis=1) + reads.drop(ctrl_labels, axis=1, inplace=True) + ctrl_label_new = ';'.join(ctrl_labels) + reads[ctrl_label_new] = ctrl_sum + except: + print(reads[ctrl_labels].sum(axis=1)) + print("Invalid input controls") + sys.exit(1) + + numClones, numColumns = reads.shape + print("Controls: " + ", ".join(ctrl_labels)) + + # + # Add pseudo count + # + + reads.iloc[:, list(range(1, numColumns))] += pseudo_count + + # + # normalize each sample to a fixed total readcount + # + sumReads = reads.iloc[:, list(range(1, numColumns))].sum(0) + normed = pd.DataFrame(index=reads.index.values) + normed['GENE'] = reads.iloc[:, 0] # first column is gene name + normed = reads.iloc[:, list(range(1, numColumns))] / np.tile(sumReads, + [numClones, 1]) * 10000000 # normalize to 10M reads + + # + # filter for minimum readcount + # + f = np.where(reads[ctrl_label_new] >= min_reads)[0] + normed = normed.iloc[f, :] + + # + # calculate fold change + # + foldchange = pd.DataFrame(index=normed.index.values) + foldchange.index.name = 'REAGENT_ID' + foldchange['GENE'] = reads.iloc[f, 0] # dataframe 'normed' has no GENE column + for i in range(numColumns - 1): + foldchange[normed.columns.values[i]] = np.log2( + (normed.loc[:, normed.columns.values[i]]) / normed[ctrl_label_new] + ) + # + # we have calculated a foldchange for the control column. Drop it. + # + foldchange.drop(ctrl_label_new, axis=1, inplace=True) + + # + # write normed readcount file + # write foldchange file + # + + foldchange_filename = output_label + '.foldchange' + foldchange.to_csv(foldchange_filename, sep='\t', float_format='%4.3f') + + normedreads_filename = output_label + '.normed_readcount' + normed.to_csv(normedreads_filename, sep='\t', float_format='%3.2f') + + +@click.command(name='bf') +@click.option('-i', '--fold-change', required=True, type=click.Path(exists=True)) +@click.option('-o', '--output-file', required=True) +@click.option('-e', '--essential-genes', required=True, type=click.Path(exists=True)) +@click.option('-n', '--non-essential-genes', required=True, type=click.Path(exists=True)) +@click.option('-c', '--columns-to-test', required=True) +@click.option('-w', '--network-file', metavar='[network File]', default=None, type=click.Path(exists=True)) +@click.option('-m', '--filter-multi-target', is_flag=True) +@click.option('-m0', '--loci-without-mismatch', type=int, default=10) +@click.option('-m1', '--loci-with-mismatch', type=int, default=10) +@click.option('--align-info', metavar='--align-info [File]', default=None, + type=click.Path(exists=True), cls=OptionRequiredIf) +@click.option('-b', '--use-bootstrapping', is_flag=True) +@click.option('-NS', '--no-resampling', is_flag=True) +@click.option('-s', '--use-small-sample', is_flag=True) +@click.option('-N', '--no-of-cross-validations', type=int, default=10) +@click.option('-NB', '--bootstrap-iterations', type=int, default=1000) +@click.option('-r', '--sgrna-bayes-factors', is_flag=True) +@click.option('-f', '--equalise-sgrna-no', type=int) +@click.option('-s', '--seed', default=int(time.time() * 100000 % 100000), type=int) +@click.option('-t', '--run-test-mode', is_flag=True) +@click.option('-p', '--equalise-rep-no', type=int) +def calculate_bayes_factors( + fold_change, output_file, essential_genes, non_essential_genes, columns_to_test, network_file, align_info, + use_bootstrapping, no_resampling, use_small_sample, filter_multi_target, loci_without_mismatch, loci_with_mismatch, + bootstrap_iterations, no_of_cross_validations, sgrna_bayes_factors, equalise_sgrna_no, seed, run_test_mode, equalise_rep_no +): + """ + \b + Calculate Bayes Factors from an input fold change file: + + \b + BAGEL.py bf -i [fold change] -o [output file] -e [essentials genes] -n [nonessentials genes] -c [columns] + + + \b + Calculates a log2 Bayes Factor for each gene. Positive BFs indicate confidence that the gene is essential. + Output written to the [output file] contains: gene name, mean Bayes Factor across all iterations, std deviation of + BFs, and number of iterations in which the gene was part of the test set (and a BF was calculated[output file]. + + + \b + Required options: + -i --fold-change [fold change file] Tab-delimited file of reagents and fold changes + (see documentation for format). + -o, --output-file [output file] Output filename + -e, --essential-genes [reference essentials] File with list of training set of essential genes + -n, --non-essential-genes [reference nonessentials] File with list of training set of nonessential genes + -c [columns to test] comma-delimited list of columns in input file to + include in analyisis + + \b + Network options: + -w [network file] Enable Network boosting. Tab-delmited file of edges. [GeneA (\\t) GeneB]\n' + + \b + Multi-target guides filtering options: + -m, --filter-multi-target Enable filtering multi-targeting guide RNAs + --align-info [file] Input precalculated align-info file + -m0, --loci-without-mismatch Filtering guide RNAs without mismatch targeting over than [N] loci, default = 10 + -m1, --loci-with-mismatch Filtering guide RNAs with 1-bp mismatch targeting over than [N] loci, default = 10 + + \b + Other options: + -b, --bootstrapping Use bootstrapping instead of cross-validation (Slow) + -NS, --no-resampling Run BAGEL without resampling + -s, --small-sample Low-fat BAGEL, Only resampled training set (Bootstrapping, iteration = 100) + -r --sgrna-bayes-factors Calculate sgRNA-wise Bayes Factor + -f --equalise-sgrna-no Equalize the number of sgRNAs per gene to particular value [Number] + -p --equalise-rep-no Equalize the number of repicates to particular value [Number] + -N --no-of-cross-validations Number of sections for cross validation (default 10) + -NB --bootstraps-iterations Number of bootstrap iterations (default 1000) + -s, --seed=N Define random seed + -h, --help Show this help text + + \b + Example: + + \b + BAGEL.py bf -i fc_file.txt -o results.bf -e ess_training_set.txt -n noness_training_set.txt -c 1,2,3 + + """ + np.random.seed(seed) # set random seed + if network_file: + network_boost = True + else: + network_boost = False + + if sgrna_bayes_factors: + rna_level = True + else: + rna_level = False + + if network_file and sgrna_bayes_factors: + network_boost = False + + if equalise_sgrna_no: + flat_sgrna = True + else: + flat_sgrna = False + + if equalise_rep_no: + flat_rep = True + else: + flat_rep = False + + if use_small_sample: + train_method = 0 + bootstrap_iterations = 100 + + elif use_bootstrapping: + train_method = 0 + + else: + train_method = 1 + + genes = {} + fc = {} + gene2rna = {} + rna2gene = {} + + multi_targeting_sgrnas = dict() + multi_targeting_sgrnas_info = dict() + + if filter_multi_target: + + try: + aligninfo = pd.read_csv(align_info, header=None, index_col=0, sep="\t").fillna("") + for seqid in aligninfo.index: + perfectmatch = 0 + mismatch_1bp = 0 + perfectmatch_gene = 0 + mismatch_1bp_gene = 0 + if aligninfo[1][seqid] != "": + perfectmatch = len(aligninfo[1][seqid].split(",")) + if aligninfo[2][seqid] != "": + perfectmatch_gene = len(aligninfo[2][seqid].split(",")) + if aligninfo[3][seqid] != "": + mismatch_1bp = len(aligninfo[3][seqid].split(",")) + if aligninfo[4][seqid] != "": + mismatch_1bp_gene = len(aligninfo[4][seqid].split(",")) + if perfectmatch > loci_without_mismatch or mismatch_1bp > loci_with_mismatch: + multi_targeting_sgrnas[seqid] = True + elif perfectmatch > 1 or mismatch_1bp > 0: + multi_targeting_sgrnas_info[seqid] = ( + perfectmatch, mismatch_1bp, perfectmatch_gene, mismatch_1bp_gene + ) + + except: + print("Please check align-info file") + sys.exit(1) + + print("Total %d multi-targeting gRNAs are discarded" % len(multi_targeting_sgrnas)) + + # + # LOAD FOLDCHANGES + # + rnatagset = set() + with open(fold_change) as fin: + fieldname = fin.readline().rstrip().split('\t') + # + # DEFINE CONTROLS + # + columns = columns_to_test.split(',') + try: + try: + column_list = list(map(int, columns)) + column_labels = [fieldname[x + 1] for x in column_list] + except ValueError: + column_labels = columns + column_list = [x for x in range(len(fieldname) - 1) if + fieldname[x + 1] in column_labels] # +1 because of First column start 2 + print("Using column: " + ", ".join(column_labels)) + # print "Using column: " + ", ".join(map(str,column_list)) + + except: + print("Invalid columns") + sys.exit(1) + + for line in fin: + fields = line.rstrip().split('\t') + rnatag = fields[0] + if filter_multi_target is True: # multitargeting sgrna filtering + if rnatag in multi_targeting_sgrnas: + continue # skip multitargeting sgrna. + if rnatag in rnatagset: + print("Error! sgRNA tag duplicates") + sys.exit(1) + rnatagset.add(rnatag) + gsym = fields[1] + + genes[gsym] = 1 + if gsym not in gene2rna: + gene2rna[gsym] = [] + gene2rna[gsym].append(rnatag) + rna2gene[rnatag] = gsym + fc[rnatag] = {} + for i in column_list: + fc[rnatag][i] = float(fields[i + 1]) # per user docs, GENE is column 0, first data column is col 1. + + genes_array = np.array(list(genes.keys())) + gene_idx = np.arange(len(genes)) + print("Number of unique genes: " + str(len(genes))) + + # + # DEFINE REFERENCE SETS + # + coreEss = [] + + with open(essential_genes) as fin: + skip_header = fin.readline() + for line in fin: + coreEss.append(line.rstrip().split('\t')[0]) + coreEss = np.array(coreEss) + print("Number of reference essentials: " + str(len(coreEss))) + + nonEss = [] + with open(non_essential_genes) as fin: + skip_header = fin.readline() + for line in fin: + nonEss.append(line.rstrip().split('\t')[0]) + + nonEss = np.array(nonEss) + print("Number of reference nonessentials: " + str(len(nonEss))) + + # + # LOAD NETWORK + # + + if network_boost is True: + network = {} + edgecount = 0 + with open(network_file) as fin: + for line in fin: + linearray = line.rstrip().split('\t') # GeneA \t GeneB format + if linearray[0] in genes_array and linearray[1] in genes_array: + for i in [0, 1]: + if linearray[i] not in network: + network[linearray[i]] = {} + network[linearray[i]][linearray[-1 * (i - 1)]] = 1 # save edge information + edgecount += 1 + + print("Number of network edges: " + str(edgecount)) + + # + # INITIALIZE BFS + # + + # Define foldchange dynamic threshold. logarithm decay. + # Parameters are defined by regression (achilles data) 2**-7 was used in previous version. + + FC_THRESH = 2 ** (-1.1535 * np.log(len(np.intersect1d(genes_array, + nonEss)) + 13.324) + 0.7728) + bf = {} + boostedbf = {} + for g in genes_array: + for rnatag in gene2rna[g]: + bf[rnatag] = [] + + boostedbf[g] = [] # boosted bf at gene level + + # + # TRAINING + # + if use_small_sample: + # declare training class + # training_data = Training(setdiff1d(gene_idx,np.where(in1d(genes_array,coreEss))),cvnum=NUMCV) + # declare training class (only for Gold-standard gene set) + training_data = Training(np.where(np.in1d(genes_array, np.union1d(coreEss, nonEss)))[0], + cvnum=no_of_cross_validations) + # all non-goldstandards + all_non_gs = np.where(np.logical_not(np.in1d(genes_array, np.union1d(coreEss, nonEss))))[0] + else: + training_data = Training(gene_idx, cvnum=no_of_cross_validations) # declare training class + + if train_method == 0: + LOOPCOUNT = bootstrap_iterations + elif train_method == 1: + LOOPCOUNT = no_of_cross_validations # 10-folds + + if run_test_mode == True: + fp = open(output_file + ".traininfo", "w") + fp.write("#1: Loopcount\n#2: Training set\n#3: Testset\n") + # No resampling option + if no_resampling == True: + print("# Caution: Resampling is disabled") + LOOPCOUNT = 1 + + print("Iter TrainEss TrainNon TestSet") + sys.stdout.flush() + for loop in range(LOOPCOUNT): + currentbf = {} + printstr = "" + printstr += str(loop) + + # + # bootstrap resample (10-folds cross-validation) from gene list to get the training set + # test set for this iteration is everything not selected in bootstrap resampled (10-folds cross-validation) + # training set + # define essential and nonessential training sets: arrays of indexes + # + if no_resampling == True: + # no resampling + gene_train_idx = gene_idx + gene_test_idx = gene_idx + else: + # CV or bootstrapping + gene_train_idx, gene_test_idx = training_data.get_data(train_method) + if use_small_sample: + # test set is union of rest of training set (gold-standard) and the other genes (all of non-gold-standard) + gene_test_idx = np.union1d(gene_test_idx, all_non_gs) + + if run_test_mode: + fp.write( + "%d\n%s\n%s\n" % (loop, ",".join(genes_array[gene_train_idx]), ",".join(genes_array[gene_test_idx]))) + + train_ess = np.where(np.in1d(genes_array[gene_train_idx], coreEss))[0] + train_non = np.where(np.in1d(genes_array[gene_train_idx], nonEss))[0] + printstr += " " + str(len(train_ess)) + printstr += " " + str(len(train_non)) + printstr += " " + str(len(gene_test_idx)) + print(printstr) + sys.stdout.flush() + # + # define ess_train: vector of observed fold changes of essential genes in training set + # + ess_train_fc_list_of_lists = [fc[rnatag] for g in genes_array[gene_train_idx[train_ess]] for rnatag in + gene2rna[g]] + ess_train_fc_flat_list = [obs for sublist in ess_train_fc_list_of_lists for obs in list(sublist.values())] + # + # define non_train vector of observed fold changes of nonessential genes in training set + # + non_train_fc_list_of_lists = [fc[rnatag] for g in genes_array[gene_train_idx[train_non]] for rnatag in + gene2rna[g]] + non_train_fc_flat_list = [obs for sublist in non_train_fc_list_of_lists for obs in list(sublist.values())] + # + # calculate empirical fold change distributions for both + # + kess = stats.gaussian_kde(ess_train_fc_flat_list) + knon = stats.gaussian_kde(non_train_fc_flat_list) + # + # define empirical upper and lower bounds within which to calculate BF = f(fold change) + # + x = np.arange(-10, 2, 0.01) + nonfitx = knon.evaluate(x) + # define lower bound empirical fold change threshold: minimum FC np.where knon is above threshold + f = np.where(nonfitx > FC_THRESH) + xmin = round_to_hundredth(min(x[f])) + # define upper bound empirical fold change threshold: minimum value of log2(ess/non) + subx = np.arange(xmin, max(x[f]), 0.01) + logratio_sample = np.log2(kess.evaluate(subx) / knon.evaluate(subx)) + f = np.where(logratio_sample == logratio_sample.min()) + xmax = round_to_hundredth(subx[f]) + # + # round foldchanges to nearest 0.01 + # precalculate logratios and build lookup table (for speed) + # + logratio_lookup = {} + for i in np.arange(xmin, xmax + 0.01, 0.01): + logratio_lookup[np.around(i * 100)] = np.log2(kess.evaluate(i) / knon.evaluate(i)) + # + # calculate BFs from lookup table for withheld test set + # + + # liner interpolation + testx = list() + testy = list() + + for g in genes_array[gene_train_idx]: + for rnatag in gene2rna[g]: + for foldchange in list(fc[rnatag].values()): + if foldchange >= xmin and foldchange <= xmax: + testx.append(np.around(foldchange * 100) / 100) + testy.append(logratio_lookup[np.around(foldchange * 100)][0]) + try: + slope, intercept, r_value, p_value, std_err = stats.linregress(np.array(testx), np.array(testy)) + except: + print("Regression failed. Check quality of the screen") + sys.exit(1) + # + # BF calculation + # + + for g in genes_array[gene_test_idx]: + for rnatag in gene2rna[g]: + bayes_factor = [] + for rep in column_list: + bayes_factor.append(slope * fc[rnatag][rep] + intercept) + bf[rnatag].append(bayes_factor) + + if run_test_mode == True: + fp.close() + + num_obs = dict() + if rna_level is False: + bf_mean = dict() + bf_std = dict() + bf_norm = dict() # sgRNA number complement + if rna_level or filter_multi_target: + bf_mean_rna_rep = dict() + bf_std_rna_rep = dict() + # bf_norm_rna_rep = dict() + + for g in gene2rna: + num_obs[g] = len(bf[gene2rna[g][0]]) + if rna_level or filter_multi_target: + for rnatag in gene2rna[g]: + bf_mean_rna_rep[rnatag] = dict() + bf_std_rna_rep[rnatag] = dict() + t = list(zip(*bf[rnatag])) + for rep in range(len(column_list)): + bf_mean_rna_rep[rnatag][column_list[rep]] = np.mean(t[rep]) + bf_std_rna_rep[rnatag][column_list[rep]] = np.std(t[rep]) + + if rna_level == False: + sumofbf_list = list() + for i in range(num_obs[g]): + sumofbf = 0.0 + for rnatag in gene2rna[g]: + sumofbf += sum(bf[rnatag][i]) + sumofbf_list.append(sumofbf) # append each iter + bf_mean[g] = np.mean(sumofbf_list) + bf_std[g] = np.std(sumofbf_list) + + # + # BUILD MULTIPLE REGRESSION MODEL FOR MULTI TARGETING GUIDE RNAs + # + if filter_multi_target: + count = 0 + trainset = dict() + bf_multi_corrected_gene = dict() + bf_multi_corrected_rna = dict() + for gene in gene2rna: + # multi_targeting_sgrnas_info[seqid] = (perfectmatch, mismatch_1bp, perfectmatch_gene, mismatch_1bp_gene) + multitarget = list() + onlytarget = list() + for seqid in gene2rna[gene]: + if seqid not in aligninfo.index: + continue + if seqid in multi_targeting_sgrnas_info: + multitarget.append(seqid) + else: + onlytarget.append(seqid) + + if len(onlytarget) > 0: # comparsion between sgRNAs targeting one locus and multiple loci + if len(multitarget) > 0: + + bf_only = np.mean([sum(list(bf_mean_rna_rep[seqid].values())) for seqid in onlytarget]) + for seqid in onlytarget: + trainset[seqid] = [1, 0, 0] + + for seqid in multitarget: + if multi_targeting_sgrnas_info[seqid][2] > 1 or multi_targeting_sgrnas_info[seqid][ + 3] > 0: # train model using multi-targeting only targeting one protein coding gene + continue + + count += 1 + increment = sum(list(bf_mean_rna_rep[seqid].values())) - bf_only + + trainset[seqid] = [multi_targeting_sgrnas_info[seqid][0], multi_targeting_sgrnas_info[seqid][1], + increment] + + if count < 10: + print("Not enough train set for calculating multi-targeting effect.\n") + print("It may cause due to unmatched gRNA names between the foldchange file and the align info file.\n") + print("Filtering is not finished\n") + filter_multi_target = False + + else: + + trainset = pd.DataFrame().from_dict(trainset).T + X = trainset[[0, 1]] + y = trainset[2] + + regressor = LinearRegression() + regressor.fit(X, y) + coeff_df = pd.DataFrame(regressor.coef_, X.columns, columns=['Coefficient']) + for i in [0, 1]: + if coeff_df['Coefficient'][i] < 0: + print("Regression coefficient is below than zero. Substituted to zero\n") + coeff_df['Coefficient'][i] = 0.0 + print("Multiple effects from perfect matched loci = %.3f and 1bp mis-matched loci = %.3f" % ( + coeff_df['Coefficient'][0], coeff_df['Coefficient'][1])) + + if rna_level == False: + for g in gene2rna: + penalty = 0.0 + for seqid in gene2rna[g]: + if seqid in multi_targeting_sgrnas_info: + penalty += float(multi_targeting_sgrnas_info[seqid][0] - 1) * coeff_df['Coefficient'][ + 0] + float(multi_targeting_sgrnas_info[seqid][1]) * coeff_df['Coefficient'][1] + bf_multi_corrected_gene[g] = bf_mean[g] - penalty + else: + for g in gene2rna: + for seqid in gene2rna[g]: + if seqid in multi_targeting_sgrnas_info: + penalty = float(multi_targeting_sgrnas_info[seqid][0] - 1) * coeff_df['Coefficient'][ + 0] + float(multi_targeting_sgrnas_info[seqid][1]) * coeff_df['Coefficient'][1] + else: + penalty = 0.0 + bf_multi_corrected_rna[seqid] = sum(list(bf_mean_rna_rep[seqid].values())) - penalty + + # + # NORMALIZE sgRNA COUNT + # + if rna_level is False and flat_sgrna == True: + if filter_multi_target == True: + targetbf = bf_multi_corrected_gene + else: + targetbf = bf_mean + + for g in gene2rna: + multiple_factor = equalise_sgrna_no / float(len(gene2rna[g])) + bf_norm[g] = targetbf[g] * multiple_factor + + ''' + if bf_std[rnatag] == 0.0: + bf_norm[rnatag] = float('inf') + else: + bf_norm[g] = ( bf[rnatag] - bf_mean[rnatag] ) / bf_std[rnatag] + ''' + training_data = Training(gene_idx) # set training class reset + + # + # calculate network scores + # + + if network_boost == True and rna_level == False: # Network boost is only working for gene level + if run_test_mode == True: # TEST MODE + fp = open(output_file + ".netscore", "w") + print("\nNetwork score calculation start\n") + + networkscores = {} + for g in genes_array[gene_idx]: + if g in network: + templist = list() + for neighbor in network[g]: + if neighbor in bf_mean: + templist.append(bf_mean[neighbor]) + + templist.sort(reverse=True) + + networkscores[g] = fibo_weighted_sum(templist) + # + # start training + # + + for loop in range(LOOPCOUNT): + currentnbf = {} + printstr = "" + printstr += str(loop) + + # + # draw train, test sets + # + gene_train_idx, gene_test_idx = training_data.get_data(train_method) + # + # define essential and nonessential training sets: arrays of indexes + # + train_ess = np.where(np.in1d(genes_array[gene_train_idx], coreEss))[0] + train_non = np.where(np.in1d(genes_array[gene_train_idx], nonEss))[0] + printstr += " " + str(len(train_ess)) + printstr += " " + str(len(train_non)) + printstr += " " + str(len(gene_test_idx)) + + sys.stdout.flush() + # + # calculate Network BF for test set + # + ess_ns_list = [networkscores[x] for x in genes_array[gene_train_idx[train_ess]] if x in networkscores] + non_ns_list = [networkscores[x] for x in genes_array[gene_train_idx[train_non]] if x in networkscores] + + kess = stats.gaussian_kde(ess_ns_list) + knon = stats.gaussian_kde(non_ns_list) + # + # set x boundary for liner regression + # + testx = list() + testy = list() + xmin = float(np.inf) + xmax = float(-np.inf) + + for networkscore in np.arange(max(ess_ns_list), min(ess_ns_list), -0.01): + density_ess = kess.evaluate(networkscore)[0] + density_non = knon.evaluate(networkscore)[0] + if density_ess == 0.0 or density_non == 0.0: + continue + + if np.log2(density_ess / density_non) > -5 and networkscore < np.array(ess_ns_list).mean(): # reverse + xmin = min(xmin, networkscore) + + for networkscore in np.arange(min(non_ns_list), max(non_ns_list), 0.01): + density_ess = kess.evaluate(networkscore)[0] + density_non = knon.evaluate(networkscore)[0] + if density_ess == 0.0 or density_non == 0.0: + continue + if np.log2(density_ess / density_non) < 5 and networkscore > np.array(non_ns_list).mean(): # reverse + xmax = max(xmax, networkscore) + # + # liner regression + # + testx = list() + testy = list() + for g in genes_array[gene_train_idx]: + if g in networkscores: + if networkscores[g] >= xmin and networkscores[g] <= xmax: + testx.append(np.around(networkscores[g] * 100) / 100) + testy.append(np.log2(kess.evaluate(networkscores[g])[0] / knon.evaluate(networkscores[g])[0])) + + slope, intercept, r_value, p_value, std_err = stats.linregress(np.array(testx), np.array(testy)) + + for g in genes_array[gene_test_idx]: + if g in networkscores: + if run_test_mode == True: + fp.write("%s\t%f\t%f\n" % (g, networkscores[g], slope * networkscores[g] + intercept)) + nbf = slope * networkscores[g] + intercept + else: + nbf = 0.0 + + boostedbf[g].append(bf_mean[g] + nbf) + if flat_sgrna == True: + boostedbf[g].append(bf_norm[g] + nbf) + + if run_test_mode == True: + fp.close() + + + + + # + # print out results + # + + # Equalizing factor (Replicates) + if flat_rep==True: + eqf = equalise_rep_no/float(len(column_labels)) + else: + eqf = 1 + + # print out + with open(output_file, 'w') as fout: + + if rna_level == True: + fout.write('RNA\tGENE') + for i in range(len(column_list)): + fout.write('\t{0:s}'.format(column_labels[i])) + if train_method == 0: + fout.write('\t{0:s}'.format(column_labels[i] + "_STD")) + fout.write('\tBF') + if train_method == 0: + fout.write('\tNumObs') + fout.write('\n') + + for rnatag in sorted(bf.keys()): + # RNA tag + fout.write('{0:s}\t'.format(rnatag)) + # Gene + gene = rna2gene[rnatag] + fout.write('{0:s}\t'.format(gene)) + + # BF of replicates + for rep in column_list: + fout.write('{0:4.3f}\t'.format(bf_mean_rna_rep[rnatag][rep])) + if train_method == 0: + fout.write('{0:4.3f}\t'.format(bf_std_rna_rep[rnatag][rep])) + + # Sum BF of replicates + if filter_multi_target == True: + fout.write('{0:4.3f}'.format(float(bf_multi_corrected_rna[rnatag]) * eqf)) # eqf = equalizing factor for the number of replicates + else: + fout.write('{0:4.3f}'.format(float(sum(list(bf_mean_rna_rep[rnatag].values()))) * eqf)) + + # Num obs + if train_method == 0: + fout.write('\t{0:d}'.format(num_obs[gene])) + fout.write('\n') + else: + fout.write('GENE') + if network_boost == True: + fout.write('\tBoostedBF') + if train_method == 0: + fout.write('\tSTD_BoostedBF') + fout.write('\tBF') + if train_method == 0: + fout.write('\tSTD\tNumObs') + if flat_sgrna == True: + fout.write('\tNormBF') + fout.write('\n') + + for g in sorted(genes.keys()): + # Gene + fout.write('{0:s}'.format(g)) + if network_boost == True: + boostedbf_mean = np.mean(boostedbf[g]) + boostedbf_std = np.std(boostedbf[g]) + fout.write('\t{0:4.3f}'.format(float(boostedbf_mean) * eqf)) + if train_method == 0: + fout.write('\t{0:4.3f}'.format(float(boostedbf_std) * eqf)) + + # BF + if filter_multi_target == True: + fout.write('\t{0:4.3f}'.format(float(bf_multi_corrected_gene[g]) * eqf)) # eqf = equalizing factor for the number of replicates + else: + fout.write('\t{0:4.3f}'.format(float(bf_mean[g]) * eqf )) + # STD, Count + if train_method == 0: + fout.write('\t{0:4.3f}\t{1:d}'.format(float(bf_std[g]), num_obs[g])) + # Normalized BF + if flat_sgrna == True: + fout.write('\t{0:4.3f}'.format(float(bf_norm[g]))) + + fout.write('\n') + + +@click.command(name='pr') +@click.option('-i', '--bayes-factors', required=True, + type=click.Path(exists=True)) +@click.option('-o', '--output-file', required=True) +@click.option('-e', '--essential-genes', required=True, + type=click.Path(exists=True)) +@click.option('-n', '--non-essential-genes', required=True, type=click.Path(exists=True)) +@click.option('-k', '--use-column', default=None) +def calculate_precision_recall(bayes_factors, output_file, essential_genes, non_essential_genes, use_column): + """ + Calculate precision-recall from an input Bayes Factors file: + + \b + BAGEL.py pr -i [Bayes Factor file] -o [output file] -e [essentials genes] -n [nonessentials genes] + + \b + Required options: + -i, --bayes-factors [Bayes factors] BAGEL output file. + -o, --output-file [output file] Output filename + -e, --essential-genes [reference essentials] File with list of training set of essential genes + -n, --non-essential-genes [reference nonessentials] File with list of training set of nonessential genes + + \b + Other options: + -k [column name] Use other column (default \BF\) + + \b + Example: + BAGEL.py pr -i input.bf -o output.PR -e ref_essentials.txt -n ref_nonessentials.txt + + """ + # + # test for availability of all files + # + essentials = pd.read_csv(essential_genes, index_col=0, sep="\t") + nonessentials = pd.read_csv(non_essential_genes, index_col=0, sep="\t") + bf = pd.read_csv(bayes_factors, index_col=0, sep="\t") + + if use_column is not None: + bf_column = use_column + if bf_column not in bf.dtypes.index: + print("Error! the column name is not in the file") + sys.exit(1) + else: + bf_column = 'BF' + + bf.sort_values(by=bf_column, ascending=False, inplace=True) + + cumulative_tp = 0. + cumulative_fp = 0. + precision = 1. + recall = 0. + # note float formats + + ess = essentials.index.values + non = nonessentials.index.values + totNumEssentials = len([x for x in bf.index.values if x in ess]) + + with open(output_file, 'w') as fout: + + fout.write('Gene\t') + fout.write(bf_column) + fout.write('\tRecall\tPrecision\tFDR\n') + + for g in bf.index.values: + if (g in ess): + cumulative_tp += 1 + elif (g in non): + cumulative_fp += 1 + recall = cumulative_tp / totNumEssentials + if ((cumulative_tp > 0) | (cumulative_fp > 0)): + precision = cumulative_tp / (cumulative_tp + cumulative_fp) + fout.write('{0:s}\t{1:4.3f}\t{2:4.3f}\t{3:4.3f}\t{4:4.3f}\n'.format(g, bf.loc[g, bf_column], recall, precision, 1.0-precision)) + + +if __name__ == '__main__': + bagel.add_command(calculate_fold_change) + bagel.add_command(calculate_bayes_factors) + bagel.add_command(calculate_precision_recall) + bagel.add_command(report_bagel_version) + bagel() diff --git a/modules/local/bagel2_fc.nf b/modules/local/bagel2_fc.nf new file mode 100644 index 00000000..9fe4ccc9 --- /dev/null +++ b/modules/local/bagel2_fc.nf @@ -0,0 +1,57 @@ +// 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 BAGEL2_FC { + tag "${meta.treatment}_${meta.reference}" + 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 "python=3.11.4 pandas=2.0.3 numpy=1.25.1 scikit-learn=1.3.0 click=8.1.6" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/mulled-v2-1ec3f69e7819b1ab3e6f57d16594eb40ed7d6792:f94a27287a1921ce0dacd411d48acff738d3ca90-0': + 'biocontainers/mulled-v2-1ec3f69e7819b1ab3e6f57d16594eb40ed7d6792:f94a27287a1921ce0dacd411d48acff738d3ca90-0' }" + + input: + // TODO nf-core: Where applicable all sample-specific information e.g. "id", "single_end", "read_group" + // MUST be provided as an input via a Groovy Map called "meta". + // This information may not be required in some instances e.g. indexing reference genome files: + // https://github.com/nf-core/modules/blob/master/modules/nf-core/bwa/index/main.nf + // TODO nf-core: Where applicable please provide/convert compressed files as input/output + // e.g. "*.fastq.gz" and NOT "*.fastq", "*.bam" and NOT "*.sam" etc. + tuple val(meta), path(count_table) + + output: + tuple val(meta), path("*.foldchange"), emit: foldchange + tuple val(meta), path("*.normed_readcount"), emit: normed_counts + //path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + + """ + BAGEL.py fc -i $count_table -o ${meta.treatment}_${meta.reference} -c $meta.reference $args + + """ + +} diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index ed8f28aa..45280efa 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -14,7 +14,7 @@ def checkPathParamList = [ params.multiqc_config, params.reference_fasta, params for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } } // Check mandatory parameters -if (!params.count_table) { ch_input = file(params.input) } else { error('Input samplesheet not specified!') } +//if (!params.count_table) { ch_input = file(params.input) } else { error('Input samplesheet not specified!') } if (params.library) { ch_library = file(params.library) } if (params.crisprcleanr) { ch_crisprcleanr= Channel.value(params.crisprcleanr) } @@ -60,6 +60,8 @@ include { MAGECK_MLE } from '../modules/nf-core/mageck/mle/main include { MAGECK_TEST } from '../modules/nf-core/mageck/test/main' include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main' include { CRISPRCLEANR_NORMALIZE } from '../modules/nf-core/crisprcleanr/normalize/main' +include { BAGEL2_FC } from '../modules/local/bagel2_fc' + /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ RUN MAIN WORKFLOW @@ -138,7 +140,7 @@ workflow CRISPRSEQ_SCREENING { if(params.rra_contrasts) { Channel.fromPath(params.rra_contrasts) - .splitCsv(header:true, sep:',' ) + .splitCsv(header:true, sep:';' ) .set { ch_contrasts } counts = ch_contrasts.combine(ch_counts) @@ -147,6 +149,18 @@ workflow CRISPRSEQ_SCREENING { ) } + if(params.rra_contrasts) { + Channel.fromPath(params.rra_contrasts) + .splitCsv(header:true, sep:';' ) + .set { ch_bagel } + counts = ch_bagel.combine(ch_counts) + + BAGEL2_FC ( + counts + ) + } + + if(params.mle_design_matrix) { ch_mle = ch_counts.combine(ch_design) ch_mle.map { From ac2ca2c09651bcb3e5ffbee5fa0d4eb6349c7f01 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Thu, 27 Jul 2023 16:17:16 +0200 Subject: [PATCH 025/133] Trying to add BAGEL_BF --- assets/CEGv2.txt | 685 +++++++++++++++++++++++++++++++ assets/NEGv1.txt | 1 + assets/schema_input.json | 4 +- modules/local/bagel2_bf.nf | 33 ++ modules/local/bagel2_fc.nf | 27 -- nextflow.config | 2 + nextflow_schema.json | 23 +- workflows/crisprseq_screening.nf | 30 +- 8 files changed, 762 insertions(+), 43 deletions(-) create mode 100644 assets/CEGv2.txt create mode 100644 assets/NEGv1.txt create mode 100644 modules/local/bagel2_bf.nf diff --git a/assets/CEGv2.txt b/assets/CEGv2.txt new file mode 100644 index 00000000..fa3ce335 --- /dev/null +++ b/assets/CEGv2.txt @@ -0,0 +1,685 @@ +GENE HGNC_ID ENTREZ_ID +AARS HGNC:20 16 +ABCE1 HGNC:69 6059 +ABCF1 HGNC:70 23 +ACTB HGNC:132 60 +ACTL6A HGNC:24124 86 +ACTR10 HGNC:17372 55860 +ACTR2 HGNC:169 10097 +ADSL HGNC:291 158 +ADSS HGNC:292 159 +AHCY HGNC:343 191 +ALG1 HGNC:18294 56052 +ALG14 HGNC:28287 199857 +ALG2 HGNC:23159 85365 +ANAPC2 HGNC:19989 29882 +ANAPC4 HGNC:19990 29945 +ANAPC5 HGNC:15713 51433 +AQR HGNC:29513 9716 +ARCN1 HGNC:649 372 +ARIH1 HGNC:689 25820 +ARL2 HGNC:693 402 +ATP2A2 HGNC:812 488 +ATP5F1A HGNC:823 498 +ATP5F1B HGNC:830 506 +ATP5F1C HGNC:833 509 +ATP5F1D HGNC:837 513 +ATP5J2-PTCD1 NA +ATP5MG HGNC:14247 10632 +ATP5PO HGNC:850 539 +ATP6V0B HGNC:861 533 +ATP6V0C HGNC:855 527 +ATP6V1A HGNC:851 523 +ATP6V1D HGNC:13527 51382 +ATP6V1E1 HGNC:857 529 +ATR HGNC:882 545 +AURKB HGNC:11390 9212 +BANF1 HGNC:17397 8815 +BIRC5 HGNC:593 332 +BUB1B HGNC:1149 701 +BUB3 HGNC:1151 9184 +BUD31 HGNC:29629 8896 +BYSL HGNC:1157 705 +TWNK HGNC:1160 56652 +C1orf109 HGNC:26039 54955 +CFAP298 HGNC:1301 56683 +NEPRO HGNC:24496 25871 +SPOUT1 HGNC:26933 51490 +CCDC84 HGNC:30460 338657 +YJU2 HGNC:25518 55702 +CCNA2 HGNC:1578 890 +CCNH HGNC:1594 902 +CCNK HGNC:1596 8812 +CCT2 HGNC:1615 10576 +CCT3 HGNC:1616 7203 +CCT4 HGNC:1617 10575 +CCT5 HGNC:1618 22948 +CCT6A HGNC:1620 908 +CCT7 HGNC:1622 10574 +CCT8 HGNC:1623 10694 +CDC123 HGNC:16827 8872 +CDC16 HGNC:1720 8881 +CDC20 HGNC:1723 991 +CDC27 HGNC:1728 996 +CDC37 HGNC:1735 11140 +CDC5L HGNC:1743 988 +CDC73 HGNC:16783 79577 +CDK1 HGNC:1722 983 +CDK7 HGNC:1778 1022 +CDK9 HGNC:1780 1025 +CDT1 HGNC:24576 81620 +CEBPZ HGNC:24218 10153 +CENPA HGNC:1851 1058 +CENPC NA +CFL1 HGNC:1874 1072 +CHAF1A HGNC:1910 10036 +CHAF1B HGNC:1911 8208 +CHEK1 HGNC:1925 1111 +CHERP HGNC:16930 10523 +CHMP2A HGNC:30216 27243 +CHMP6 HGNC:25675 79643 +CIAO1 HGNC:14280 9391 +CINP HGNC:23789 51550 +UTP4 HGNC:1983 84916 +CKAP5 HGNC:28959 9793 +CLNS1A HGNC:2080 1207 +CLP1 HGNC:16999 10978 +CLTC HGNC:2092 1213 +CMPK1 HGNC:18170 51727 +CMTR1 NA +CNOT3 HGNC:7879 4849 +COA5 HGNC:33848 493753 +COPA HGNC:2230 1314 +COPB1 HGNC:2231 1315 +COPB2 HGNC:2232 9276 +COPS3 HGNC:2239 8533 +COPS6 HGNC:21749 10980 +COPZ1 HGNC:2243 22818 +COQ4 HGNC:19693 51117 +COX10 HGNC:2260 1352 +COX11 HGNC:2261 1353 +COX15 HGNC:2263 1355 +COX4I1 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+DNM2 HGNC:2974 1785 +DNMT1 HGNC:2976 1786 +DOLK HGNC:23406 22845 +DONSON HGNC:2993 29980 +DPAGT1 HGNC:2995 1798 +DTL HGNC:30288 51514 +DTYMK HGNC:3061 1841 +DYNC1I2 HGNC:2964 1781 +ECD HGNC:17029 11319 +EEF2 HGNC:3214 1938 +EFTUD2 HGNC:30858 9343 +EIF2B1 HGNC:3257 1967 +EIF2B3 HGNC:3259 8891 +EIF2B5 HGNC:3261 8893 +EIF2S1 HGNC:3265 1965 +EIF2S2 HGNC:3266 8894 +EIF2S3 HGNC:3267 1968 +EIF3A HGNC:3271 8661 +EIF3B HGNC:3280 8662 +EIF3C HGNC:3279 8663 +EIF3D HGNC:3278 8664 +EIF3G HGNC:3274 8666 +EIF3I HGNC:3272 8668 +EIF4A3 HGNC:18683 9775 +EIF5A HGNC:3300 1984 +EIF5B HGNC:30793 9669 +EIF6 HGNC:6159 3692 +ELAC2 HGNC:14198 60528 +ELL HGNC:23114 8178 +EPRS HGNC:3418 2058 +ERCC2 HGNC:3434 2068 +ERCC3 HGNC:3435 2071 +ERH HGNC:3447 2079 +EXOSC2 HGNC:17097 23404 +EXOSC3 HGNC:17944 51010 +EXOSC4 HGNC:18189 54512 +EXOSC6 HGNC:19055 118460 +EXOSC7 HGNC:28112 23016 +EXOSC8 HGNC:17035 11340 +CIAO2B HGNC:24261 51647 +FARS2 HGNC:21062 10667 +FARSA HGNC:3592 2193 +FARSB HGNC:17800 10056 +FAU HGNC:3597 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at master · hart-lab/bagel","locale":"en"} \ No newline at end of file diff --git a/assets/schema_input.json b/assets/schema_input.json index 21c8c38a..dfc3dc1c 100644 --- a/assets/schema_input.json +++ b/assets/schema_input.json @@ -1,5 +1,5 @@ { - "$schema": "http://json-schema.org/draft-07/schema", + "$schema": "http://json-schema.org/draft-07/schema", "$id": "https://raw.githubusercontent.com/nf-core/crisprseq/master/assets/schema_input.json", "title": "nf-core/crisprseq pipeline - params.input schema", "description": "Schema for the file provided with params.input", @@ -30,8 +30,6 @@ "condition": { "type": "string", "pattern": "^\\S+$", - "enum": ["control", "treatment"], - "errorMessage": "Condition name must be one of 'control' or 'treatment'", "meta": ["condition"] }, "reference": { diff --git a/modules/local/bagel2_bf.nf b/modules/local/bagel2_bf.nf new file mode 100644 index 00000000..5299915a --- /dev/null +++ b/modules/local/bagel2_bf.nf @@ -0,0 +1,33 @@ +process BAGEL2_BF { + tag "$meta.treatment" + label 'process_single' + + + conda "python=3.11.4 pandas=2.0.3 numpy=1.25.1 scikit-learn=1.3.0 click=8.1.6" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/mulled-v2-1ec3f69e7819b1ab3e6f57d16594eb40ed7d6792:f94a27287a1921ce0dacd411d48acff738d3ca90-0': + 'biocontainers/mulled-v2-1ec3f69e7819b1ab3e6f57d16594eb40ed7d6792:f94a27287a1921ce0dacd411d48acff738d3ca90-0' }" + + + input: + tuple val(meta), path(foldchange) + path(reference_essentials) + path(reference_nonessentials) + + output: + tuple val(meta), path("*.pr"), emit: pr + //path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.treatment}" + + """ + BAGEL.py bf -i $foldchange -o ${meta.treatment} $args -e $reference_essentials -n $reference_nonessentials -c ${meta.treatment} + + """ + +} diff --git a/modules/local/bagel2_fc.nf b/modules/local/bagel2_fc.nf index 9fe4ccc9..bc0c4f12 100644 --- a/modules/local/bagel2_fc.nf +++ b/modules/local/bagel2_fc.nf @@ -1,40 +1,13 @@ -// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) -// https://github.com/nf-core/modules/tree/master/modules/nf-core/ -// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: -// https://nf-co.re/join -// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. -// All other parameters MUST be provided using the "task.ext" directive, see here: -// https://www.nextflow.io/docs/latest/process.html#ext -// where "task.ext" is a string. -// Any parameters that need to be evaluated in the context of a particular sample -// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. -// TODO nf-core: Software that can be piped together SHOULD be added to separate module files -// unless there is a run-time, storage advantage in implementing in this way -// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: -// bwa mem | samtools view -B -T ref.fasta -// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty -// list (`[]`) instead of a file can be used to work around this issue. - process BAGEL2_FC { tag "${meta.treatment}_${meta.reference}" label 'process_single' - // TODO nf-core: List required Conda package(s). - // Software MUST be pinned to channel (i.e. "bioconda"), version (i.e. "1.10"). - // For Conda, the build (i.e. "h9402c20_2") must be EXCLUDED to support installation on different operating systems. - // TODO nf-core: See section in main README for further information regarding finding and adding container addresses to the section below. conda "python=3.11.4 pandas=2.0.3 numpy=1.25.1 scikit-learn=1.3.0 click=8.1.6" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? 'https://depot.galaxyproject.org/singularity/mulled-v2-1ec3f69e7819b1ab3e6f57d16594eb40ed7d6792:f94a27287a1921ce0dacd411d48acff738d3ca90-0': 'biocontainers/mulled-v2-1ec3f69e7819b1ab3e6f57d16594eb40ed7d6792:f94a27287a1921ce0dacd411d48acff738d3ca90-0' }" 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(count_table) output: diff --git a/nextflow.config b/nextflow.config index 4b01987c..bf675e5f 100644 --- a/nextflow.config +++ b/nextflow.config @@ -21,6 +21,8 @@ params { count_table = null min_reads = 30 min_targeted_genes = 3 + bagel_reference_essentials = 'https://raw.githubusercontent.com/hart-lab/bagel/master/CEGv2.txt' + bagel_reference_nonessentials = 'https://raw.githubusercontent.com/hart-lab/bagel/master/NEGv1.txt' // Pipeline steps overrepresented = false diff --git a/nextflow_schema.json b/nextflow_schema.json index c75c5a2d..c5774253 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -186,12 +186,22 @@ "min_reads": { "type": "number", "description": "a filter threshold value for sgRNAs, based on their average counts in the control sample", - "default": 30 + "default": 30.0 }, "min_targeted_genes": { "type": "number", "description": "Minimal number of different genes targeted by sgRNAs in a biased segment in order for the corresponding counts to be corrected for CRISPRcleanR", - "default": 3 + "default": 3.0 + }, + "bagel_reference_essentials": { + "type": "string", + "description": "Core essential gene set for BAGEL2", + "default": "https://github.com/hart-lab/bagel/blob/master/CEGv2.txt" + }, + "bagel_reference_nonessentials": { + "type": "string", + "description": "Non essential gene set for BAGEL2", + "default": "https://github.com/hart-lab/bagel/blob/master/NEGv1.txt" } } }, @@ -450,21 +460,12 @@ { "$ref": "#/definitions/umi_parameters" }, - { - "$ref": "#/definitions/targeted_pipeline_steps" - }, - { - "$ref": "#/definitions/umi_parameters" - }, { "$ref": "#/definitions/targeted_parameters" }, { "$ref": "#/definitions/vsearch_parameters" }, - { - "$ref": "#/definitions/vsearch_parameters" - }, { "$ref": "#/definitions/screening_parameters" }, diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index e0e9e478..129c1229 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -67,6 +67,7 @@ include { MAGECK_TEST } from '../modules/nf-core/mageck/test/mai include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main' include { CRISPRCLEANR_NORMALIZE } from '../modules/nf-core/crisprcleanr/normalize/main' include { BAGEL2_FC } from '../modules/local/bagel2_fc' +include { BAGEL2_BF } from '../modules/local/bagel2_bf' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -85,17 +86,19 @@ workflow CRISPRSEQ_SCREENING { // // Create input channel from input file provided through params.input // + single_end = true Channel.fromSamplesheet("input") .map{ meta, fastq_1, fastq_2, x, y, z -> // x (reference), y (protospacer), and z (template) are part of the targeted workflows and we don't need them if (!fastq_2) { + single_end = true return [ meta, [ fastq_1 ] ] } else { + single_end = false return [ meta, [ fastq_1, fastq_2 ] ] } } .set { ch_input } - // // MODULE: Run FastQC // @@ -104,6 +107,7 @@ workflow CRISPRSEQ_SCREENING { ) ch_versions = ch_versions.mix(FASTQC.out.versions.first()) + if(single_end==true) { ch_input .map { meta, fastq -> [meta.condition, fastq] @@ -116,6 +120,10 @@ workflow CRISPRSEQ_SCREENING { } .set { joined } + joined.dump(tag: "input joined") + } + + // // MODULE: Run mageck count // @@ -167,12 +175,30 @@ workflow CRISPRSEQ_SCREENING { .set { ch_bagel } counts = ch_bagel.combine(ch_counts) + if(!params.bagel_reference_essentials) { + ch_bagel_reference_essentials = Channel.fromPath("${projectDir}/assets/CEGv2.txt", checkIfExists: true) + } + + ch_bagel_reference_essentials.dump(tag: "input joined") + + if(!params.bagel_reference_nonessentials) { + ch_bagel_reference_nonessentials = Channel.fromPath("${projectDir}/assets/NEGv1.txt", checkIfExists: true) + } + + BAGEL2_FC ( - counts + counts, + ch_bagel_reference_essentials, + ch_bagel_reference_nonessentials ) } + //BAGEL2_BF ( + // BAGEL2_FC.out.foldchange + //) + + if(params.mle_design_matrix) { ch_mle = ch_counts.combine(ch_design) ch_mle.map { From 8a888035d1b9197ae4b8ce0e1880c7ac25e670ea Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Fri, 28 Jul 2023 16:01:57 +0200 Subject: [PATCH 026/133] Adding bagel2_bf --- assets/NEGv1.txt | 930 ++++++++++++++++++++++++++++++- modules/local/bagel2_bf.nf | 2 +- modules/local/bagel2_fc.nf | 2 +- workflows/crisprseq_screening.nf | 29 +- 4 files changed, 950 insertions(+), 13 deletions(-) diff --git a/assets/NEGv1.txt b/assets/NEGv1.txt index 32701540..96401816 100644 --- a/assets/NEGv1.txt +++ b/assets/NEGv1.txt @@ -1 +1,929 @@ 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388364 +TMPRSS11A HGNC:27954 339967 +TMPRSS11B HGNC:25398 132724 +TMPRSS11F HGNC:29994 389208 +TMPRSS12 HGNC:28779 283471 +TMPRSS15 HGNC:9490 5651 +TNR HGNC:11953 7143 +TPD52L3 HGNC:23382 89882 +TPH2 HGNC:20692 121278 +TPRX1 HGNC:32174 284355 +TPTE HGNC:12023 7179 +TREML4 HGNC:30807 285852 +TRHR HGNC:12299 7201 +TRIM40 HGNC:18736 135644 +TRIM42 HGNC:19014 287015 +TRIM43 HGNC:19015 129868 +TRIM48 HGNC:19021 79097 +TRIM49 HGNC:13431 57093 +TRIM51 HGNC:19023 84767 +TRIM60 HGNC:21162 166655 +TRIM67 HGNC:31859 440730 +TRIML1 HGNC:26698 339976 +TRPC5 HGNC:12337 7224 +TRPC7 HGNC:20754 57113 +TRPM1 HGNC:7146 4308 +TRPV5 HGNC:3145 56302 +TSGA13 HGNC:12369 114960 +TSHB HGNC:12372 7252 +TSPAN16 HGNC:30725 26526 +TSPO2 HGNC:21256 222642 +TSPY1 HGNC:12381 7258 +TSPYL6 HGNC:14521 388951 +TSSK1B HGNC:14968 83942 +TSSK2 HGNC:11401 23617 +TXNDC8 HGNC:31454 255220 +TYR HGNC:12442 7299 +UBQLN3 HGNC:12510 50613 +UMOD HGNC:12559 7369 +UROC1 HGNC:26444 131669 +USP17L2 HGNC:34434 377630 +USP26 HGNC:13485 83844 +USP29 HGNC:18563 57663 +UTS2R HGNC:4468 2837 +VAX1 HGNC:12660 11023 +VCX3A HGNC:18159 51481 +VHLL HGNC:30666 391104 +VN1R2 HGNC:19872 317701 +VN1R4 HGNC:19871 317703 +VN1R5 HGNC:19870 317705 +VPREB1 HGNC:12709 7441 +VRTN HGNC:20223 55237 +VSX2 HGNC:1975 338917 +WFDC10A HGNC:16139 140832 +WFDC11 HGNC:20478 259239 +WFDC9 HGNC:20380 259240 +XAGE2 HGNC:4112 9502 +XAGE5 HGNC:30930 170627 +XKR7 HGNC:23062 343702 +ZAN HGNC:12857 7455 +ZCCHC13 HGNC:31749 389874 +RTL4 HGNC:25214 340595 +ZG16 HGNC:30961 653808 +ZIC3 HGNC:12874 7547 +ZIM3 HGNC:16366 114026 +CBLL2 HGNC:26371 158506 +ZNF648 HGNC:18190 127665 +ZNF679 HGNC:28650 168417 +ZNF804B HGNC:21958 219578 +ZNRF4 HGNC:17726 148066 +ZP2 HGNC:13188 7783 +ZP4 HGNC:15770 57829 +ZSWIM2 HGNC:30990 151112 + diff --git a/modules/local/bagel2_bf.nf b/modules/local/bagel2_bf.nf index 5299915a..56fb66a0 100644 --- a/modules/local/bagel2_bf.nf +++ b/modules/local/bagel2_bf.nf @@ -26,7 +26,7 @@ process BAGEL2_BF { def prefix = task.ext.prefix ?: "${meta.treatment}" """ - BAGEL.py bf -i $foldchange -o ${meta.treatment} $args -e $reference_essentials -n $reference_nonessentials -c ${meta.treatment} + BAGEL.py bf -i $foldchange -o '${meta.treatment}.pr' $args -e $reference_essentials -n $reference_nonessentials -c ${meta.treatment} """ diff --git a/modules/local/bagel2_fc.nf b/modules/local/bagel2_fc.nf index bc0c4f12..bec12be1 100644 --- a/modules/local/bagel2_fc.nf +++ b/modules/local/bagel2_fc.nf @@ -23,7 +23,7 @@ process BAGEL2_FC { def prefix = task.ext.prefix ?: "${meta.id}" """ - BAGEL.py fc -i $count_table -o ${meta.treatment}_${meta.reference} -c $meta.reference $args + BAGEL.py fc -i $count_table -o ${meta.treatment}_vs_${meta.reference} -c $meta.reference $args """ diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 129c1229..8b060b9f 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -175,28 +175,37 @@ workflow CRISPRSEQ_SCREENING { .set { ch_bagel } counts = ch_bagel.combine(ch_counts) + //MAKE THIS PRETTIER if(!params.bagel_reference_essentials) { - ch_bagel_reference_essentials = Channel.fromPath("${projectDir}/assets/CEGv2.txt", checkIfExists: true) - } + ch_bagel_reference_essentials = Channel.fromPath("${projectDir}/assets/CEGv2.txt") + } else { + ch_bagel_reference_essentials = Channel.fromPath(params.bagel_reference_essentials) + } ch_bagel_reference_essentials.dump(tag: "input joined") if(!params.bagel_reference_nonessentials) { - ch_bagel_reference_nonessentials = Channel.fromPath("${projectDir}/assets/NEGv1.txt", checkIfExists: true) - } + ch_bagel_reference_nonessentials = Channel.fromPath("${projectDir}/assets/NEGv1.txt") + } else { + ch_bagel_reference_nonessentials = Channel.fromPath(params.bagel_reference_nonessentials) } BAGEL2_FC ( - counts, - ch_bagel_reference_essentials, - ch_bagel_reference_nonessentials + counts ) } + BAGEL2_FC.out.foldchange.dump(tag: "BAGEL2_FC dump") + + test = ch_bagel_reference_essentials.combine(BAGEL2_FC.out.foldchange) + test.dump(tag: "TEST") - //BAGEL2_BF ( - // BAGEL2_FC.out.foldchange - //) + + BAGEL2_BF ( + BAGEL2_FC.out.foldchange, + ch_bagel_reference_essentials, + ch_bagel_reference_nonessentials + ) if(params.mle_design_matrix) { From 03826f5e9e2de4522709102ef80775085a6a58e5 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 31 Jul 2023 09:20:38 +0200 Subject: [PATCH 027/133] add all module version --- workflows/crisprseq_targeted.nf | 12 ++++++++++++ 1 file changed, 12 insertions(+) diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 53c28942..2a3a5427 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -305,6 +305,7 @@ workflow CRISPRSEQ_TARGETED { return [ meta, reads[0], adapter_seqs[0] ] } .set { ch_adapter_seqs } + ch_versions = ch_versions.mix(FIND_ADAPTERS.out.versions.first()) // @@ -376,6 +377,7 @@ workflow CRISPRSEQ_TARGETED { EXTRACT_UMIS ( SEQTK_SEQ_MASK.out.fastx ) + ch_versions = ch_versions.mix(EXTRACT_UMIS.out.versions.first()) // @@ -384,6 +386,7 @@ workflow CRISPRSEQ_TARGETED { VSEARCH_CLUSTER ( EXTRACT_UMIS.out.fasta ) + ch_versions = ch_versions.mix(VSEARCH_CLUSTER.out.versions.first()) // Obtain a file with UBS (UBI bin size) and UMI ID VSEARCH_CLUSTER.out.clusters @@ -434,6 +437,7 @@ workflow CRISPRSEQ_TARGETED { ch_umi_bysize.cluster, Channel.value("--sortbysize") ) + ch_versions = ch_versions.mix(VSEARCH_SORT.out.versions.first()) // Get the correspondent fasta sequencences from top cluster sequences // Replaces the sequence name adding the "centroid_" prefix to avoid having two sequences with the same name in following steps @@ -496,6 +500,7 @@ workflow CRISPRSEQ_TARGETED { false, false ) + ch_versions = ch_versions.mix(MINIMAP2_ALIGN_UMI_1.out.versions.first()) // Only continue with clusters that have aligned sequences @@ -511,6 +516,7 @@ workflow CRISPRSEQ_TARGETED { .join(ch_top_clusters_sequence) .join(ch_minimap_1) ) + ch_versions = ch_versions.mix(RACON_1.out.versions.first()) // // MODULE: Mapping with minimap2 - cycle 2 @@ -523,6 +529,7 @@ workflow CRISPRSEQ_TARGETED { false, false ) + ch_versions = ch_versions.mix(MINIMAP2_ALIGN_UMI_2.out.versions.first()) // Only continue with clusters that have aligned sequences MINIMAP2_ALIGN_UMI_2.out.paf @@ -537,6 +544,7 @@ workflow CRISPRSEQ_TARGETED { .join(RACON_1.out.improved_assembly) .join(ch_minimap_2) ) + ch_versions = ch_versions.mix(RACON_2.out.versions.first()) // @@ -546,6 +554,7 @@ workflow CRISPRSEQ_TARGETED { ch_clusters_sequence .join(RACON_2.out.improved_assembly) ) + ch_versions = ch_versions.mix(MEDAKA.out.versions.first()) // Collect all consensus UMI sequences into one single file per sample MEDAKA.out.assembly @@ -577,6 +586,7 @@ workflow CRISPRSEQ_TARGETED { SEQTK_SEQ_FATOFQ ( ch_umi_consensus ) + ch_versions = ch_versions.mix(SEQTK_SEQ_FATOFQ.out.versions.first()) } ch_preprocess_reads = params.umi_clustering ? SEQTK_SEQ_FATOFQ.out.fastx : SEQTK_SEQ_MASK.out.fastx @@ -665,6 +675,7 @@ workflow CRISPRSEQ_TARGETED { ORIENT_REFERENCE.out.reference .join(ch_seq_template) ) + ch_versions = ch_versions.mix(TEMPLATE_REFERENCE.out.versions.first()) // @@ -725,6 +736,7 @@ workflow CRISPRSEQ_TARGETED { CIGAR_PARSER ( ch_to_parse_cigar ) + ch_versions = ch_versions.mix(CIGAR_PARSER.out.versions.first()) // From f5ab95de4a19021af7d56928c4d7269dd7209db9 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 31 Jul 2023 09:40:15 +0200 Subject: [PATCH 028/133] return Rscript version instead of R --- modules/local/cigar_parser.nf | 2 +- modules/local/template_reference.nf | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/cigar_parser.nf b/modules/local/cigar_parser.nf index 8c87314a..fcdce47a 100644 --- a/modules/local/cigar_parser.nf +++ b/modules/local/cigar_parser.nf @@ -39,7 +39,7 @@ process CIGAR_PARSER { cat <<-END_VERSIONS > versions.yml "${task.process}": - R: \$(R --version) + Rscript: \$(Rscript --version) END_VERSIONS """ } diff --git a/modules/local/template_reference.nf b/modules/local/template_reference.nf index 12c24553..28fcfd45 100644 --- a/modules/local/template_reference.nf +++ b/modules/local/template_reference.nf @@ -29,7 +29,7 @@ process TEMPLATE_REFERENCE { cat <<-END_VERSIONS > versions.yml "${task.process}": - R: \$(R --version) + Rscript: \$(Rscript --version) END_VERSIONS """ } From 0bebd941a03021b612b7ba4675ab9fd6a071effb Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Mon, 31 Jul 2023 10:07:31 +0200 Subject: [PATCH 029/133] working modules of bagel --- modules/local/bagel2_bf.nf | 4 +--- workflows/crisprseq_screening.nf | 11 +++++------ 2 files changed, 6 insertions(+), 9 deletions(-) diff --git a/modules/local/bagel2_bf.nf b/modules/local/bagel2_bf.nf index 56fb66a0..4d3deb6a 100644 --- a/modules/local/bagel2_bf.nf +++ b/modules/local/bagel2_bf.nf @@ -10,9 +10,7 @@ process BAGEL2_BF { input: - tuple val(meta), path(foldchange) - path(reference_essentials) - path(reference_nonessentials) + tuple val(meta), path(foldchange), path(reference_essentials), path(reference_nonessentials) output: tuple val(meta), path("*.pr"), emit: pr diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 8b060b9f..ff46746f 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -182,7 +182,7 @@ workflow CRISPRSEQ_SCREENING { ch_bagel_reference_essentials = Channel.fromPath(params.bagel_reference_essentials) } - ch_bagel_reference_essentials.dump(tag: "input joined") + //ch_bagel_reference_essentials.dump(tag: "input joined") if(!params.bagel_reference_nonessentials) { ch_bagel_reference_nonessentials = Channel.fromPath("${projectDir}/assets/NEGv1.txt") @@ -195,16 +195,15 @@ workflow CRISPRSEQ_SCREENING { ) } - BAGEL2_FC.out.foldchange.dump(tag: "BAGEL2_FC dump") - test = ch_bagel_reference_essentials.combine(BAGEL2_FC.out.foldchange) + test = BAGEL2_FC.out.foldchange.combine(ch_bagel_reference_essentials) + .combine(ch_bagel_reference_nonessentials) + test.dump(tag: "TEST") BAGEL2_BF ( - BAGEL2_FC.out.foldchange, - ch_bagel_reference_essentials, - ch_bagel_reference_nonessentials + test ) From 64e2fa12975f5636d647101435cb77bef6a0bf3f Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 31 Jul 2023 10:07:33 +0200 Subject: [PATCH 030/133] remove module samtools faidx --- modules.json | 5 --- modules/nf-core/samtools/faidx/main.nf | 44 ----------------------- modules/nf-core/samtools/faidx/meta.yml | 47 ------------------------- workflows/crisprseq_targeted.nf | 1 - 4 files changed, 97 deletions(-) delete mode 100644 modules/nf-core/samtools/faidx/main.nf delete mode 100644 modules/nf-core/samtools/faidx/meta.yml diff --git a/modules.json b/modules.json index 22939a8d..e396008c 100644 --- a/modules.json +++ b/modules.json @@ -102,11 +102,6 @@ "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", "installed_by": ["modules"] }, - "samtools/faidx": { - "branch": "master", - "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", - "installed_by": ["modules"] - }, "samtools/index": { "branch": "master", "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", diff --git a/modules/nf-core/samtools/faidx/main.nf b/modules/nf-core/samtools/faidx/main.nf deleted file mode 100644 index 4dd0e5b0..00000000 --- a/modules/nf-core/samtools/faidx/main.nf +++ /dev/null @@ -1,44 +0,0 @@ -process SAMTOOLS_FAIDX { - tag "$fasta" - label 'process_single' - - conda "bioconda::samtools=1.17" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : - 'biocontainers/samtools:1.17--h00cdaf9_0' }" - - input: - tuple val(meta), path(fasta) - - output: - tuple val(meta), path ("*.fai"), emit: fai - tuple val(meta), path ("*.gzi"), emit: gzi, optional: true - path "versions.yml" , emit: versions - - when: - task.ext.when == null || task.ext.when - - script: - def args = task.ext.args ?: '' - """ - samtools \\ - faidx \\ - $args \\ - $fasta - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') - END_VERSIONS - """ - - stub: - """ - touch ${fasta}.fai - cat <<-END_VERSIONS > versions.yml - - "${task.process}": - samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') - END_VERSIONS - """ -} diff --git a/modules/nf-core/samtools/faidx/meta.yml b/modules/nf-core/samtools/faidx/meta.yml deleted file mode 100644 index fe2fe9a1..00000000 --- a/modules/nf-core/samtools/faidx/meta.yml +++ /dev/null @@ -1,47 +0,0 @@ -name: samtools_faidx -description: Index FASTA file -keywords: - - index - - fasta -tools: - - samtools: - description: | - SAMtools is a set of utilities for interacting with and post-processing - short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. - These files are generated as output by short read aligners like BWA. - homepage: http://www.htslib.org/ - documentation: http://www.htslib.org/doc/samtools.html - doi: 10.1093/bioinformatics/btp352 - licence: ["MIT"] -input: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - fasta: - type: file - description: FASTA file - pattern: "*.{fa,fasta}" -output: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - fai: - type: file - description: FASTA index file - pattern: "*.{fai}" - - gzi: - type: file - description: Optional gzip index file for compressed inputs - pattern: "*.gzi" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" -authors: - - "@drpatelh" - - "@ewels" - - "@phue" diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 2a3a5427..ed1a56af 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -75,7 +75,6 @@ include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_ORIGINAL } from '../modules/nf-core/m include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_UMI_1 } from '../modules/nf-core/minimap2/align/main' include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_UMI_2 } from '../modules/nf-core/minimap2/align/main' include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_TEMPLATE } from '../modules/nf-core/minimap2/align/main' -include { SAMTOOLS_FAIDX } from '../modules/nf-core/samtools/faidx/main' include { MINIMAP2_INDEX } from '../modules/nf-core/minimap2/index/main' include { MEDAKA } from '../modules/nf-core/medaka/main' include { CUTADAPT } from '../modules/nf-core/cutadapt/main' From a6d40903775ab4fbd7741a53ad6c00602dab729f Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 1 Aug 2023 14:40:47 +0200 Subject: [PATCH 031/133] All of the bagel modules and output graphs --- modules/local/{bagel2_bf.nf => bagel2/bf.nf} | 4 +- modules/local/{bagel2_fc.nf => bagel2/fc.nf} | 0 modules/local/bagel2/graph.nf | 77 ++++++++++++++++++++ modules/local/bagel2/pr.nf | 30 ++++++++ workflows/crisprseq_screening.nf | 27 +++++-- 5 files changed, 128 insertions(+), 10 deletions(-) rename modules/local/{bagel2_bf.nf => bagel2/bf.nf} (89%) rename modules/local/{bagel2_fc.nf => bagel2/fc.nf} (100%) create mode 100644 modules/local/bagel2/graph.nf create mode 100644 modules/local/bagel2/pr.nf diff --git a/modules/local/bagel2_bf.nf b/modules/local/bagel2/bf.nf similarity index 89% rename from modules/local/bagel2_bf.nf rename to modules/local/bagel2/bf.nf index 4d3deb6a..1353965c 100644 --- a/modules/local/bagel2_bf.nf +++ b/modules/local/bagel2/bf.nf @@ -13,7 +13,7 @@ process BAGEL2_BF { tuple val(meta), path(foldchange), path(reference_essentials), path(reference_nonessentials) output: - tuple val(meta), path("*.pr"), emit: pr + tuple val(meta), path("*.bf"), emit: bf //path "versions.yml" , emit: versions when: @@ -24,7 +24,7 @@ process BAGEL2_BF { def prefix = task.ext.prefix ?: "${meta.treatment}" """ - BAGEL.py bf -i $foldchange -o '${meta.treatment}.pr' $args -e $reference_essentials -n $reference_nonessentials -c ${meta.treatment} + BAGEL.py bf -i $foldchange -o '${meta.treatment}.bf' $args -e $reference_essentials -n $reference_nonessentials -c ${meta.treatment} """ diff --git a/modules/local/bagel2_fc.nf b/modules/local/bagel2/fc.nf similarity index 100% rename from modules/local/bagel2_fc.nf rename to modules/local/bagel2/fc.nf diff --git a/modules/local/bagel2/graph.nf b/modules/local/bagel2/graph.nf new file mode 100644 index 00000000..1bc149a4 --- /dev/null +++ b/modules/local/bagel2/graph.nf @@ -0,0 +1,77 @@ +// TODO nf-core: If in doubt look at other nf-core/modules to see how we are doing things! :) +// https://github.com/nf-core/modules/tree/master/modules/nf-core/ +// You can also ask for help via your pull request or on the #modules channel on the nf-core Slack workspace: +// https://nf-co.re/join +// TODO nf-core: A module file SHOULD only define input and output files as command-line parameters. +// All other parameters MUST be provided using the "task.ext" directive, see here: +// https://www.nextflow.io/docs/latest/process.html#ext +// where "task.ext" is a string. +// Any parameters that need to be evaluated in the context of a particular sample +// e.g. single-end/paired-end data MUST also be defined and evaluated appropriately. +// TODO nf-core: Software that can be piped together SHOULD be added to separate module files +// unless there is a run-time, storage advantage in implementing in this way +// e.g. it's ok to have a single module for bwa to output BAM instead of SAM: +// bwa mem | samtools view -B -T ref.fasta +// TODO nf-core: Optional inputs are not currently supported by Nextflow. However, using an empty +// list (`[]`) instead of a file can be used to work around this issue. + +process BAGEL2_GRAPH { + tag "$meta.id" + label 'process_single' + + conda "python=3.11.4 pandas=2.0.3 numpy=1.25.1 scikit-learn=1.3.0 click=8.1.6 matplotlib=3.7.2" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/mulled-v2-54e0353146eca1531516863e8235bf7385d76663:c9ff1a9eec871c54cbea815eae778da702623978-0': + 'biocontainers/mulled-v2-54e0353146eca1531516863e8235bf7385d76663:c9ff1a9eec871c54cbea815eae778da702623978-0' }" + + + input: + tuple val(meta), path(pr) + + output: + //path "versions.yml" , emit: versions + path("*.png") , emit: pictures + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + + """ + #!/usr/bin/env python3 + + import pandas as pd + import matplotlib.pyplot as plt + + pr_data = pd.read_table('${pr}', index_col=0) + + plt.plot(pr_data.Recall, pr_data.Precision, linewidth=2) + plt.xlim(0, 1.01) + plt.ylim(0, 1.02) + plt.xlabel('Recall') + plt.ylabel('Precision (1-FDR)') + plt.title('Precision-Recall Plot') + + # Save the plot to a PNG file + file_name = 'PR_plot_{}_vs_{}.png'.format('${meta.treatment}', '${meta.reference}') + + plt.savefig(file_name) + + # Show the plot (optional) + plt.show() + + pr_data.hist('BF', bins=50, range=(-100,100)) + plt.xlabel('Bayes Factor') + plt.ylabel('Number of Genes') + + file_name = 'barplot_{}_vs_{}.png'.format('${meta.treatment}', '${meta.reference}') + + plt.savefig(file_name) + plt.show() + + """ + + +} diff --git a/modules/local/bagel2/pr.nf b/modules/local/bagel2/pr.nf new file mode 100644 index 00000000..4a5616b4 --- /dev/null +++ b/modules/local/bagel2/pr.nf @@ -0,0 +1,30 @@ +process BAGEL2_PR { + tag "$meta.treatment" + label 'process_single' + + conda "python=3.11.4 pandas=2.0.3 numpy=1.25.1 scikit-learn=1.3.0 click=8.1.6" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/mulled-v2-1ec3f69e7819b1ab3e6f57d16594eb40ed7d6792:f94a27287a1921ce0dacd411d48acff738d3ca90-0': + 'biocontainers/mulled-v2-1ec3f69e7819b1ab3e6f57d16594eb40ed7d6792:f94a27287a1921ce0dacd411d48acff738d3ca90-0' }" + + + input: + tuple val(meta), path(bf), path(reference_essentials), path(reference_nonessentials) + + output: + tuple val(meta), path("*.pr") , emit: pr + + //path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" + + """ + BAGEL.py pr -i $bf -o '${meta.treatment}.pr' -e $reference_essentials -n $reference_nonessentials + """ + +} diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index ff46746f..317838d2 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -66,8 +66,10 @@ include { MAGECK_MLE } from '../modules/nf-core/mageck/mle/main include { MAGECK_TEST } from '../modules/nf-core/mageck/test/main' include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main' include { CRISPRCLEANR_NORMALIZE } from '../modules/nf-core/crisprcleanr/normalize/main' -include { BAGEL2_FC } from '../modules/local/bagel2_fc' -include { BAGEL2_BF } from '../modules/local/bagel2_bf' +include { BAGEL2_FC } from '../modules/local/bagel2/fc' +include { BAGEL2_BF } from '../modules/local/bagel2/bf' +include { BAGEL2_PR } from '../modules/local/bagel2/pr' +include { BAGEL2_GRAPH } from '../modules/local/bagel2/graph' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -175,7 +177,7 @@ workflow CRISPRSEQ_SCREENING { .set { ch_bagel } counts = ch_bagel.combine(ch_counts) - //MAKE THIS PRETTIER + //TO DO MAKE THIS PRETTIER if(!params.bagel_reference_essentials) { ch_bagel_reference_essentials = Channel.fromPath("${projectDir}/assets/CEGv2.txt") } else { @@ -184,6 +186,7 @@ workflow CRISPRSEQ_SCREENING { //ch_bagel_reference_essentials.dump(tag: "input joined") + //TO DO make this prettier if(!params.bagel_reference_nonessentials) { ch_bagel_reference_nonessentials = Channel.fromPath("${projectDir}/assets/NEGv1.txt") } else { @@ -195,17 +198,25 @@ workflow CRISPRSEQ_SCREENING { ) } - - test = BAGEL2_FC.out.foldchange.combine(ch_bagel_reference_essentials) + ch_bagel_bf = BAGEL2_FC.out.foldchange.combine(ch_bagel_reference_essentials) .combine(ch_bagel_reference_nonessentials) - test.dump(tag: "TEST") - BAGEL2_BF ( - test + ch_bagel_bf ) + BAGEL2_BF.out.bf.dump(tag:"BAGEL2_BF dump") + ch_bagel_pr = BAGEL2_BF.out.bf.combine(ch_bagel_reference_essentials) + .combine(ch_bagel_reference_nonessentials) + + BAGEL2_PR ( + ch_bagel_pr + ) + BAGEL2_PR.out.pr.dump(tag:"TEST PR DUMP") + BAGEL2_GRAPH ( + BAGEL2_PR.out.pr + ) if(params.mle_design_matrix) { ch_mle = ch_counts.combine(ch_design) From 26c8df37c901a7a660c38e643964df097aee79a9 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Wed, 2 Aug 2023 10:43:31 +0200 Subject: [PATCH 032/133] add edition plot to multiQC --- assets/multiqc_config.yml | 48 +++++++++++++++++++++++++++++++++ bin/cigar_parser.R | 30 ++++++++------------- docs/output/targeted.md | 2 +- workflows/crisprseq_targeted.nf | 1 + 4 files changed, 61 insertions(+), 20 deletions(-) diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 73d174a7..c7fc0616 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -11,3 +11,51 @@ report_section_order: order: -1002 export_plots: true + +# Custom tables and plots +custom_data: + # Summary of edition plot + edition_plot: + plot_type: "bargraph" + file_format: "csv" + section_name: "RESULTS: Type of edition" + description: | + Number of reads classified according to edition types. + Reads are classified between WT (without an edit), template based modifications and indels. + Indels are divided between deletions, insertions and delins (deletion + insertion). + Deletions and insertions can be out of frame or in frame. + categories: + Wt: + name: "WT reads" + color: "#C0C0C0" + Template-based: + name: "Template-based modifications" + color: "#E9AF82" + Delins: + name: "DelIns" + color: "#D5B1F5" + Ins_inframe: + name: "Insertions in frame" + color: "#B5D5C1" + Ins_outframe: + name: "Insertions out of frame" + color: "#80D09F" + Dels_inframe: + name: "Deletions in frame" + color: "#BFD0EB" + Dels_outframe: + name: "Deletions out of frame" + color: "#699AE9" + pconfig: + id: "edition_bargraph" + title: "Reads by edition type" + ylab: "Read count" +sp: + edition_plot: + fn: "*_edits.csv" + +# Define the order of sections +# Show FastQC before Custom results +module_order: + - fastqc + - custom_content diff --git a/bin/cigar_parser.R b/bin/cigar_parser.R index 7c2179da..2da1fd9e 100755 --- a/bin/cigar_parser.R +++ b/bin/cigar_parser.R @@ -1088,7 +1088,7 @@ if (dim(alignment_info)[1] != 0){ ##### Summary: edition ########### - edit_classes <- c("Wt", "Template-based", "Indels", "Insertions", "Deletions", "Delins", "Dels inframe", "Dels outfarme", "Ins inframe", "Ins outfarme") + edit_classes <- c("Wt", "Template-based", "Indels", "Insertions", "Deletions", "Delins", "Dels_inframe", "Dels_outframe", "Ins_inframe", "Ins_outframe") ##### Update wt if template-based is a substitution if ( t_type == "subs"){ @@ -1098,21 +1098,11 @@ if (dim(alignment_info)[1] != 0){ edit_counts <- c(wt_reads, t_reads, indels_count, ins_count, dels_count, delin_count, in_frame_dels, out_frame_dels, in_frame_ins, out_frame_ins ) edit_summary <- data.frame(classes = unlist(edit_classes), counts = unlist(edit_counts)) - total_reads <- wt_reads+t_reads+indels_count - wt_perc <- (wt_reads/total_reads)*100 - temp_perc <- ((t_reads)/total_reads)*100 - indels_perc <- (indels_count/total_reads)*100 - ins_perc <- (ins_count/indels_count)*100 - dels_perc <- (dels_count/indels_count)*100 - delins_perc <- (delin_count/indels_count)*100 - in_frame_ins_perc <- (in_frame_ins/ins_count)*100 - out_frame_ins_perc <- (out_frame_ins/ins_count)*100 - in_frame_dels_perc <- (in_frame_dels/dels_count)*100 - out_frame_dels_perc <- (out_frame_dels/dels_count)*100 - - edit_classes_perc <- c("Wt", "Template-based", "Indels", "Delins", "Insertions", "Ins inframe", "Ins outfarme", "Deletions", "Dels inframe", "Dels outfarme") - edit_counts_perc <- c(round(wt_perc,2), round(temp_perc,2), round(indels_perc,2), round(delins_perc,2), round(ins_perc,2), round(in_frame_ins_perc,2), round(out_frame_ins_perc,2), round(dels_perc,2), round(in_frame_dels_perc,2), round(out_frame_dels_perc,2 )) - edit_summary_perc <- data.frame(classes = unlist(edit_classes_perc), counts = unlist(edit_counts_perc)) + edit_classes_perc <- c("Wt", "Template-based", "Delins", "Ins_inframe", "Ins_outframe", "Dels_inframe", "Dels_outframe") + edit_counts_perc <- c(wt_reads, t_reads, delin_count, in_frame_ins, out_frame_ins, in_frame_dels, out_frame_dels) + edit_summary_perc <- data.frame(sample = unlist(edit_counts_perc), row.names = unlist(edit_classes_perc)) + colnames(edit_summary_perc)[1] = results_path # Rename the column to add the sample ID + edit_summary_perc <- t(edit_summary_perc) # t() will add classes as columns and counts as values, 1 row per sample ### Save edits count write.csv(edit_summary_perc,file=paste0(results_path, "_edits.csv")) @@ -1201,8 +1191,10 @@ if (dim(alignment_info)[1] != 0){ separated_indels = data.frame(matrix(nrow = 1, ncol = length(columns))) colnames(separated_indels) = columns write.csv(separated_indels,file=paste0(results_path, "_Badlyindels.csv")) - edit_classes_perc <- c("Wt", "Template-based", "Indels", "Delins", "Insertions", "Ins inframe", "Ins outfarme", "Deletions", "Dels inframe", "Dels outfarme") - edit_counts_perc <- c(0, 0, 0, 0, 0, 0, 0, 0, 0, 0) - edit_summary_perc <- data.frame(classes = unlist(edit_classes_perc), counts = unlist(edit_counts_perc)) + edit_classes_perc <- c("Wt", "Template-based", "Delins", "Ins_inframe", "Ins_outframe", "Dels_inframe", "Dels_outframe") + edit_counts_perc <- c(0, 0, 0, 0, 0, 0, 0) + edit_summary_perc <- data.frame(sample = unlist(edit_counts_perc), row.names = unlist(edit_classes_perc)) + colnames(edit_summary_perc)[1] = results_path# Rename the column to add the sample ID + edit_summary_perc <- t(edit_summary_perc) # t() will add classes as columns and counts as values, 1 row per sample write.csv(edit_summary_perc,file=paste0(results_path, "_edits.csv")) } diff --git a/docs/output/targeted.md b/docs/output/targeted.md index b04ca3d8..5aa68b3e 100644 --- a/docs/output/targeted.md +++ b/docs/output/targeted.md @@ -252,7 +252,7 @@ This section contains the final output of the pipeline. It contains information - `*_cutSite.json`: Contains the protospacer cut site position in the reference. - `*_edition.html`: Interactive pie chart with the percentage of edition types. Reads are classified between WT (without an edit) and indels. Indels are divided between deletions, insertions and delins (deletion + insertion). Deletions and insertions can be out of frame or in frame. ![Test sample hCas9-AAVS1-a edition plot](../images/hCas9-AAVS1-a_edition.png) - - `*_edits.csv`: Table containing the data visualized in the pie chart. + - `*_edits.csv`: Table containing the number of reads classified to each edition type. Contains the data visualized in the pie chart. - `*_indels.csv`: Table containing information of all reads. Edit type, edit start and length, if the edition happens above the error rate, if it's located into the common edit window, the frequency, the percentage, the pattern, surrounding nucleotides in case of insertions, the protospacer cut site, the sample id, number of aligned reads and number of reads with and without a template modification. - `*_QC-indels.html`: Interactive pie chart with information about aligned reads. Reads are classified between WT and containing indels. Both types are classified between passing the filtering steps or not. Indel reads passing the filtering steps are divided in reads with a modification above the error rate and located in the common edit window, above the error rate but not in the edit region, vice versa, or any of those conditions. ![Test sample hCas9-AAVS1-a QC indels plot](../images/hCas9-AAVS1-a_QC-indels.png) diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index ed1a56af..511231c8 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -757,6 +757,7 @@ workflow CRISPRSEQ_TARGETED { ch_multiqc_files = Channel.empty() ch_multiqc_files = ch_multiqc_files.mix(ch_workflow_summary.collectFile(name: 'workflow_summary_mqc.yaml')) ch_multiqc_files = ch_multiqc_files.mix(ch_methods_description.collectFile(name: 'methods_description_mqc.yaml')) + ch_multiqc_files = ch_multiqc_files.mix(CIGAR_PARSER.out.edition.collect{it[2]}.ifEmpty([])) ch_multiqc_files = ch_multiqc_files.mix(CUSTOM_DUMPSOFTWAREVERSIONS.out.mqc_yml.collect()) ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}.ifEmpty([])) From 41a9eaa0c991b1f630c7b4dc36d3cfad343c4d07 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 3 Aug 2023 09:00:22 +0200 Subject: [PATCH 033/133] add indel QC plot to multiqc --- assets/multiqc_config.yml | 44 ++++++++++++++++++++++++++++++++- bin/cigar_parser.R | 22 +++++++++++++---- modules/local/cigar_parser.nf | 1 + workflows/crisprseq_targeted.nf | 1 + 4 files changed, 62 insertions(+), 6 deletions(-) diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index c7fc0616..d40cb078 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -23,7 +23,7 @@ custom_data: Number of reads classified according to edition types. Reads are classified between WT (without an edit), template based modifications and indels. Indels are divided between deletions, insertions and delins (deletion + insertion). - Deletions and insertions can be out of frame or in frame. + Deletions and insertions can be out of frame or in frame categories: Wt: name: "WT reads" @@ -50,9 +50,51 @@ custom_data: id: "edition_bargraph" title: "Reads by edition type" ylab: "Read count" + # Indels QC plot + indel_qc_plot: + plot_type: "bargraph" + file_format: "csv" + section_name: "RESULTS: QC of indels" + description: | + Number of reads classified between WT and containing indels. + Both types are classified between passing the filtering steps or not. + Indel reads passing the filtering steps are divided input: + - reads with a modification above the error rate and located in the common edit window + - above the error rate but not in the edit region + - below the error rate and within the edit region + - below the error rate and not in the the edit region + categories: + Wt NOT passing filter: + name: "WT reads not passing filtering steps" + color: "#302F2F" + Wt passing filter: + name: "WT reads passing filtering steps" + color: "#C0C0C0" + Indels NOT passing filter: + name: "Reads with indels not passing filtering steps" + color: "#696868" + Above error & in pick: + name: "Reads with indels above the error rate and within the edition region" + color: "#80D09F" + NOT above error & in pick: + name: "Reads with indels below the error rate and within the edition region" + color: "#FFB570" + Above error & NOT in pick: + name: "Reads with indels above the error rate and outside the edition region" + color: "#F89D47" + NOT above error & NOT in pick: + name: "Reads with indels below the error rate and outside the edition region" + color: "#EB5555" + pconfig: + id: "indelqc_bargraph" + title: "Read indels quality control" + ylab: "Read count" + sp: edition_plot: fn: "*_edits.csv" + indel_qc_plot: + fn: "*_QC-indels.csv" # Define the order of sections # Show FastQC before Custom results diff --git a/bin/cigar_parser.R b/bin/cigar_parser.R index 2da1fd9e..523519fd 100755 --- a/bin/cigar_parser.R +++ b/bin/cigar_parser.R @@ -1008,7 +1008,6 @@ if (dim(alignment_info)[1] != 0){ reads_classes <- c("Raw reads", "Merged reads", "Quality filtered reads", "Clustered reads", "Aligned reads") reads_counts <- c(as.character(reads), as.character(merged_reads), as.character(trimmed_reads), as.character(clustered_reads), as.character(aligned_reads)) - write(reads_counts, stdout()) reads_summary <- data.frame(classes = unlist(reads_classes), counts = unlist(reads_counts)) ########### Pre-variant calling counts @@ -1026,8 +1025,15 @@ if (dim(alignment_info)[1] != 0){ "Above error & in pick", "NOT above error & in pick", "NOT above error & NOT in pick", "Above error & NOT in pick") prevc_counts <- c(aligned_reads, wt_reads+incorrect_wt, dim(separated_indels)[1]+trunc_reads, wt_reads, incorrect_wt, dim(separated_indels)[1], trunc_reads, ep, nep, nenp, enp) - write(prevc_counts, stdout()) prevc_summary <- data.frame(classes = unlist(prevc_classes), counts = unlist(prevc_counts)) + # Write prevc_summary data to a csv for multiQC plots + prevc_classes_mqc <- c("Wt passing filter", "Wt NOT passing filter", "Indels NOT passing filter", + "Above error & in pick", "NOT above error & in pick", "NOT above error & NOT in pick", "Above error & NOT in pick") + prevc_counts_mqc <- c(wt_reads, incorrect_wt, trunc_reads, ep, nep, nenp, enp) + indel_filters <- data.frame(sample = unlist(prevc_counts_mqc), row.names = unlist(prevc_classes_mqc)) + colnames(indel_filters)[1] = results_path # Rename the column to add the sample ID + indel_filters <- t(indel_filters) # t() will add classes as columns and counts as values, 1 row per sample + write.csv(indel_filters,file=paste0(results_path, "_QC-indels.csv")) } else { ###########################In case we don't have indels but maybe there are template based editions @@ -1064,7 +1070,6 @@ if (dim(alignment_info)[1] != 0){ reads_classes <- c("Raw reads", "Merged reads", "Quality filtered reads", "Clustered reads", "Aligned reads") reads_counts <- c(as.character(reads), as.character(merged_reads), as.character(trimmed_reads), as.character(clustered_reads), as.character(aligned_reads)) - write(reads_counts, stdout()) reads_summary <- data.frame(classes = unlist(reads_classes), counts = unlist(reads_counts)) ########### Pre-variant calling counts @@ -1076,12 +1081,19 @@ if (dim(alignment_info)[1] != 0){ prevc_classes <- c("Aligned reads", "Wt", "Indels", "Wt passing filter", "Wt NOT passing filter", "Indels passing filter", "Indels NOT passing filter", "Above error & in pick", "NOT above error & in pick", "NOT above error & NOT in pick", "Above error & NOT in pick") - prevc_counts <- c(aligned_reads, wt_reads+incorrect_wt, 0+trunc_reads, wt_reads, incorrect_wt, 0, trunc_reads, + prevc_counts <- c(aligned_reads, wt_reads+incorrect_wt, dim(separated_indels)[1]+trunc_reads, wt_reads, incorrect_wt, dim(separated_indels)[1], trunc_reads, ep, nep, nenp, enp) - write(prevc_counts, stdout()) prevc_summary <- data.frame(classes = unlist(prevc_classes), counts = unlist(prevc_counts)) exportJson <- toJSON(cut_site) write(exportJson, paste(sample_id,"_cutSite.json",sep = "")) + # Write prevc_summary data to a csv for multiQC plots + prevc_classe_mqc <- c("Wt passing filter", "Wt NOT passing filter", "Indels NOT passing filter", + "Above error & in pick", "NOT above error & in pick", "NOT above error & NOT in pick", "Above error & NOT in pick") + prevc_counts_mqc <- c(wt_reads, incorrect_wt, trunc_reads, ep, nep, nenp, enp) + indel_filters <- data.frame(sample = unlist(prevc_counts_mqc), row.names = unlist(prevc_classe_mqc)) + colnames(indel_filters)[1] = results_path # Rename the column to add the sample ID + indel_filters <- t(indel_filters) # t() will add classes as columns and counts as values, 1 row per sample + write.csv(indel_filters,file=paste0(results_path, "_QC-indels.csv")) } ########### diff --git a/modules/local/cigar_parser.nf b/modules/local/cigar_parser.nf index fcdce47a..d1ffa10b 100644 --- a/modules/local/cigar_parser.nf +++ b/modules/local/cigar_parser.nf @@ -14,6 +14,7 @@ process CIGAR_PARSER { tuple val(meta), path("*indels.csv"), path("*_subs-perc.csv"), emit: indels tuple val(meta), path("*.html"), path("*edits.csv") , emit: edition tuple val(meta), path("*cutSite.json") , emit: cutsite + tuple val(meta), path("*_QC-indels.csv") , emit: qcindels path "versions.yml" , emit: versions when: diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 511231c8..1814eaef 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -758,6 +758,7 @@ workflow CRISPRSEQ_TARGETED { ch_multiqc_files = ch_multiqc_files.mix(ch_workflow_summary.collectFile(name: 'workflow_summary_mqc.yaml')) ch_multiqc_files = ch_multiqc_files.mix(ch_methods_description.collectFile(name: 'methods_description_mqc.yaml')) ch_multiqc_files = ch_multiqc_files.mix(CIGAR_PARSER.out.edition.collect{it[2]}.ifEmpty([])) + ch_multiqc_files = ch_multiqc_files.mix(CIGAR_PARSER.out.qcindels.collect{it[1]}.ifEmpty([])) ch_multiqc_files = ch_multiqc_files.mix(CUSTOM_DUMPSOFTWAREVERSIONS.out.mqc_yml.collect()) ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}.ifEmpty([])) From 6f59fa0f994af081d5a7b78f397bd6b6f8222ef3 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 3 Aug 2023 09:06:00 +0200 Subject: [PATCH 034/133] remove module samtools faidx --- modules.json | 5 --- modules/nf-core/samtools/faidx/main.nf | 44 ----------------------- modules/nf-core/samtools/faidx/meta.yml | 47 ------------------------- workflows/crisprseq_targeted.nf | 1 - 4 files changed, 97 deletions(-) delete mode 100644 modules/nf-core/samtools/faidx/main.nf delete mode 100644 modules/nf-core/samtools/faidx/meta.yml diff --git a/modules.json b/modules.json index 22939a8d..e396008c 100644 --- a/modules.json +++ b/modules.json @@ -102,11 +102,6 @@ "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", "installed_by": ["modules"] }, - "samtools/faidx": { - "branch": "master", - "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", - "installed_by": ["modules"] - }, "samtools/index": { "branch": "master", "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", diff --git a/modules/nf-core/samtools/faidx/main.nf b/modules/nf-core/samtools/faidx/main.nf deleted file mode 100644 index 4dd0e5b0..00000000 --- a/modules/nf-core/samtools/faidx/main.nf +++ /dev/null @@ -1,44 +0,0 @@ -process SAMTOOLS_FAIDX { - tag "$fasta" - label 'process_single' - - conda "bioconda::samtools=1.17" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/samtools:1.17--h00cdaf9_0' : - 'biocontainers/samtools:1.17--h00cdaf9_0' }" - - input: - tuple val(meta), path(fasta) - - output: - tuple val(meta), path ("*.fai"), emit: fai - tuple val(meta), path ("*.gzi"), emit: gzi, optional: true - path "versions.yml" , emit: versions - - when: - task.ext.when == null || task.ext.when - - script: - def args = task.ext.args ?: '' - """ - samtools \\ - faidx \\ - $args \\ - $fasta - - cat <<-END_VERSIONS > versions.yml - "${task.process}": - samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') - END_VERSIONS - """ - - stub: - """ - touch ${fasta}.fai - cat <<-END_VERSIONS > versions.yml - - "${task.process}": - samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') - END_VERSIONS - """ -} diff --git a/modules/nf-core/samtools/faidx/meta.yml b/modules/nf-core/samtools/faidx/meta.yml deleted file mode 100644 index fe2fe9a1..00000000 --- a/modules/nf-core/samtools/faidx/meta.yml +++ /dev/null @@ -1,47 +0,0 @@ -name: samtools_faidx -description: Index FASTA file -keywords: - - index - - fasta -tools: - - samtools: - description: | - SAMtools is a set of utilities for interacting with and post-processing - short DNA sequence read alignments in the SAM, BAM and CRAM formats, written by Heng Li. - These files are generated as output by short read aligners like BWA. - homepage: http://www.htslib.org/ - documentation: http://www.htslib.org/doc/samtools.html - doi: 10.1093/bioinformatics/btp352 - licence: ["MIT"] -input: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - fasta: - type: file - description: FASTA file - pattern: "*.{fa,fasta}" -output: - - meta: - type: map - description: | - Groovy Map containing sample information - e.g. [ id:'test', single_end:false ] - - fai: - type: file - description: FASTA index file - pattern: "*.{fai}" - - gzi: - type: file - description: Optional gzip index file for compressed inputs - pattern: "*.gzi" - - versions: - type: file - description: File containing software versions - pattern: "versions.yml" -authors: - - "@drpatelh" - - "@ewels" - - "@phue" diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 53c28942..6556d79d 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -75,7 +75,6 @@ include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_ORIGINAL } from '../modules/nf-core/m include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_UMI_1 } from '../modules/nf-core/minimap2/align/main' include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_UMI_2 } from '../modules/nf-core/minimap2/align/main' include { MINIMAP2_ALIGN as MINIMAP2_ALIGN_TEMPLATE } from '../modules/nf-core/minimap2/align/main' -include { SAMTOOLS_FAIDX } from '../modules/nf-core/samtools/faidx/main' include { MINIMAP2_INDEX } from '../modules/nf-core/minimap2/index/main' include { MEDAKA } from '../modules/nf-core/medaka/main' include { CUTADAPT } from '../modules/nf-core/cutadapt/main' From a66fa61e2e79b68cbc18b425689f70fdeb7baa2d Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 3 Aug 2023 11:52:32 +0200 Subject: [PATCH 035/133] add read processing table --- assets/multiqc_config.yml | 66 ++++++++++++++++++++++++++++++--- bin/cigar_parser.R | 26 ++++++++++++- modules/local/cigar_parser.nf | 1 + workflows/crisprseq_targeted.nf | 1 + 4 files changed, 88 insertions(+), 6 deletions(-) diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index d40cb078..9c52320d 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -59,10 +59,10 @@ custom_data: Number of reads classified between WT and containing indels. Both types are classified between passing the filtering steps or not. Indel reads passing the filtering steps are divided input: - - reads with a modification above the error rate and located in the common edit window - - above the error rate but not in the edit region - - below the error rate and within the edit region - - below the error rate and not in the the edit region + reads with a modification above the error rate and located in the common edit window; + above the error rate but not in the edit region; + below the error rate and within the edit region; + below the error rate and not in the the edit region; categories: Wt NOT passing filter: name: "WT reads not passing filtering steps" @@ -89,15 +89,71 @@ custom_data: id: "indelqc_bargraph" title: "Read indels quality control" ylab: "Read count" + # Read processing summary table + read_processing: + id: "read_processing" + section_name: "Read processing summary" + plot_type: "table" + description: | + Table showing the number of input raw reads per sample and the percentage of reads after each processing step: + reads merged with Pear; + reads passing quality filters; + number of resulting clusters after UMI clustering of reads; + number of aligned reads + pconfig: + id: "read_processing" + namespace: "Read processing summary" + table_title: "Read processing summary" + sortRows: False + headers: + Raw reads: + title: "Raw reads" + description: "Number of input raw reads" + format: "{:,.0f}" + scale: "PuBu" + Merged reads: + title: "Merged reads" + description: "Percentage of merged reads" + suffix: "%" + max: 100 + min: 0 + scale: "RdYlGn" + Quality filtered reads: + title: "Quality reads" + description: "Percentage of reads passing the quality filter" + suffix: "%" + max: 100 + min: 0 + scale: "RdYlGn" + Clustered reads: + title: "Clusters" + description: "Number of UMI clusters after read clustering" + format: "{:,.0f}" + scale: "PuBu" + Aligned reads: + title: "Aligned reads" + description: "Percentage of aligned reads or UMI clusters" + suffix: "%" + max: 100 + min: 0 + scale: "RdYlGn" sp: edition_plot: fn: "*_edits.csv" indel_qc_plot: fn: "*_QC-indels.csv" + read_processing: + fn: "*_reads-summary.csv" # Define the order of sections -# Show FastQC before Custom results module_order: - fastqc - custom_content + +# Set the order of custom code plots and tables +custom_content: + order: + - read_processing + - edition_plot + - indel_qc_plot diff --git a/bin/cigar_parser.R b/bin/cigar_parser.R index 523519fd..6bb972ca 100755 --- a/bin/cigar_parser.R +++ b/bin/cigar_parser.R @@ -1009,6 +1009,18 @@ if (dim(alignment_info)[1] != 0){ reads_classes <- c("Raw reads", "Merged reads", "Quality filtered reads", "Clustered reads", "Aligned reads") reads_counts <- c(as.character(reads), as.character(merged_reads), as.character(trimmed_reads), as.character(clustered_reads), as.character(aligned_reads)) reads_summary <- data.frame(classes = unlist(reads_classes), counts = unlist(reads_counts)) + # Save table for multiQC + formated_counts <- c( + strsplit(as.character(reads), ' ')[[1]][2], + strsplit(strsplit(as.character(merged_reads), '(', fixed = TRUE)[[1]][2], '%')[[1]][1], + strsplit(strsplit(as.character(trimmed_reads), '(', fixed = TRUE)[[1]][2], '%')[[1]][1], + as.character(clustered_reads), + strsplit(strsplit(as.character(aligned_reads), '(', fixed = TRUE)[[1]][2], '%')[[1]][1] + ) + reads_summary_mqc <- data.frame(sample = unlist(formated_counts), row.names = unlist(reads_classes)) + colnames(reads_summary_mqc)[1] = results_path # Rename the column to add the sample ID + reads_summary_mqc <- t(reads_summary_mqc) # t() will add classes as columns and counts as values, 1 row per sample + write.csv(reads_summary_mqc,file=paste0(results_path, "_reads-summary.csv")) ########### Pre-variant calling counts # Indel filters @@ -1071,6 +1083,18 @@ if (dim(alignment_info)[1] != 0){ reads_classes <- c("Raw reads", "Merged reads", "Quality filtered reads", "Clustered reads", "Aligned reads") reads_counts <- c(as.character(reads), as.character(merged_reads), as.character(trimmed_reads), as.character(clustered_reads), as.character(aligned_reads)) reads_summary <- data.frame(classes = unlist(reads_classes), counts = unlist(reads_counts)) + # Save table for multiQC + formated_counts <- c( + strsplit(as.character(reads), ' ')[[1]][2], + strsplit(strsplit(as.character(merged_reads), '(', fixed = TRUE)[[1]][2], '%')[[1]][1], + strsplit(strsplit(as.character(trimmed_reads), '(', fixed = TRUE)[[1]][2], '%')[[1]][1], + as.character(clustered_reads), + strsplit(strsplit(as.character(aligned_reads), '(', fixed = TRUE)[[1]][2], '%')[[1]][1] + ) + reads_summary_mqc <- data.frame(sample = unlist(formated_counts), row.names = unlist(reads_classes)) + colnames(reads_summary_mqc)[1] = results_path # Rename the column to add the sample ID + reads_summary_mqc <- t(reads_summary_mqc) # t() will add classes as columns and counts as values, 1 row per sample + write.csv(reads_summary_mqc,file=paste0(results_path, "_reads-summary.csv")) ########### Pre-variant calling counts # Indel filters @@ -1206,7 +1230,7 @@ if (dim(alignment_info)[1] != 0){ edit_classes_perc <- c("Wt", "Template-based", "Delins", "Ins_inframe", "Ins_outframe", "Dels_inframe", "Dels_outframe") edit_counts_perc <- c(0, 0, 0, 0, 0, 0, 0) edit_summary_perc <- data.frame(sample = unlist(edit_counts_perc), row.names = unlist(edit_classes_perc)) - colnames(edit_summary_perc)[1] = results_path# Rename the column to add the sample ID + colnames(edit_summary_perc)[1] = results_path # Rename the column to add the sample ID edit_summary_perc <- t(edit_summary_perc) # t() will add classes as columns and counts as values, 1 row per sample write.csv(edit_summary_perc,file=paste0(results_path, "_edits.csv")) } diff --git a/modules/local/cigar_parser.nf b/modules/local/cigar_parser.nf index d1ffa10b..5073ab26 100644 --- a/modules/local/cigar_parser.nf +++ b/modules/local/cigar_parser.nf @@ -15,6 +15,7 @@ process CIGAR_PARSER { tuple val(meta), path("*.html"), path("*edits.csv") , emit: edition tuple val(meta), path("*cutSite.json") , emit: cutsite tuple val(meta), path("*_QC-indels.csv") , emit: qcindels + tuple val(meta), path("*_reads-summary.csv") , emit: processing path "versions.yml" , emit: versions when: diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 1814eaef..984dda5d 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -757,6 +757,7 @@ workflow CRISPRSEQ_TARGETED { ch_multiqc_files = Channel.empty() ch_multiqc_files = ch_multiqc_files.mix(ch_workflow_summary.collectFile(name: 'workflow_summary_mqc.yaml')) ch_multiqc_files = ch_multiqc_files.mix(ch_methods_description.collectFile(name: 'methods_description_mqc.yaml')) + ch_multiqc_files = ch_multiqc_files.mix(CIGAR_PARSER.out.processing.collect{it[1]}.ifEmpty([])) ch_multiqc_files = ch_multiqc_files.mix(CIGAR_PARSER.out.edition.collect{it[2]}.ifEmpty([])) ch_multiqc_files = ch_multiqc_files.mix(CIGAR_PARSER.out.qcindels.collect{it[1]}.ifEmpty([])) ch_multiqc_files = ch_multiqc_files.mix(CUSTOM_DUMPSOFTWAREVERSIONS.out.mqc_yml.collect()) From c682852a445b4d50583c429277e208da4b8b9f1b Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 3 Aug 2023 13:59:41 +0200 Subject: [PATCH 036/133] add cutadapt section to multiqc and sort table columns --- assets/multiqc_config.yml | 23 ++++++++++++++++++++++- workflows/crisprseq_targeted.nf | 12 ++++++++++++ 2 files changed, 34 insertions(+), 1 deletion(-) diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 9c52320d..331a476f 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -12,6 +12,12 @@ report_section_order: export_plots: true +# Modules to run +run_modules: + - custom_content + - fastqc + - cutadapt + # Custom tables and plots custom_data: # Summary of edition plot @@ -104,7 +110,6 @@ custom_data: id: "read_processing" namespace: "Read processing summary" table_title: "Read processing summary" - sortRows: False headers: Raw reads: title: "Raw reads" @@ -145,10 +150,13 @@ sp: fn: "*_QC-indels.csv" read_processing: fn: "*_reads-summary.csv" + cutadapt: + fn: "*.cutadapt.log" # Define the order of sections module_order: - fastqc + - cutadapt - custom_content # Set the order of custom code plots and tables @@ -157,3 +165,16 @@ custom_content: - read_processing - edition_plot - indel_qc_plot + +# Order of table columns +table_columns_placement: + Read processing summary: + Raw reads: 100 + Merged reads: 200 + Quality filtered reads: 300 + Clustered reads: 400 + Aligned reads: 500 + +# Add a comment to cutadapt section showing filtered reads +section_comments: + cutadapt_filtered_reads: "Cutadapt is selecting reads passing quality filters." diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 984dda5d..b1b081a6 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -342,6 +342,15 @@ workflow CRISPRSEQ_TARGETED { .join(PEAR.out.assembled, remainder: true) .join(SEQTK_SEQ_MASK.out.fastx) .join(CUTADAPT.out.log) + .map { meta, reads, assembled, masked, trimmed -> + if (assembled == null) { + assembled = file('null_a') + } + if (trimmed == "null") { + trimmed = file('null_t') + } + return [ meta, reads, assembled, masked, trimmed ] + } .set { ch_merging_summary_data } } else { ch_cat_fastq.paired @@ -762,6 +771,9 @@ workflow CRISPRSEQ_TARGETED { ch_multiqc_files = ch_multiqc_files.mix(CIGAR_PARSER.out.qcindels.collect{it[1]}.ifEmpty([])) ch_multiqc_files = ch_multiqc_files.mix(CUSTOM_DUMPSOFTWAREVERSIONS.out.mqc_yml.collect()) ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}.ifEmpty([])) + if (params.overrepresented) { + ch_multiqc_files = ch_multiqc_files.mix(CUTADAPT.out.log.collect{it[1]}.ifEmpty([])) + } MULTIQC ( ch_multiqc_files.collect(), From 54499cef8f6fda4e7bbe6e10bb51765bb2f28969 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 3 Aug 2023 14:04:10 +0200 Subject: [PATCH 037/133] update changelog --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index e5d26623..24902a8d 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -9,6 +9,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - Template update v2.9 ([#52](https://github.com/nf-core/crisprseq/pull/52)) - Use `Channel.fromSamplesheet()` from `nf-validation` to validate input sample sheets and create an input channel ([#58](https://github.com/nf-core/crisprseq/pull/58)) +- Add custom plots to MultiQC report (cutadapt module, read processing, edition, edition QC) ([#64](https://github.com/nf-core/crisprseq/pull/64)) ### Fixed From 4843f582f01b96b65088a0bf8f6a17302bf072c8 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 3 Aug 2023 14:16:50 +0200 Subject: [PATCH 038/133] update documentation about multiQC output --- docs/images/mqc_qc_of_indels.png | Bin 0 -> 84853 bytes 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z`{&~r0shHIDn2cKV)Xj|19n-kSpWb4 literal 0 HcmV?d00001 diff --git a/docs/output/targeted.md b/docs/output/targeted.md index 5aa68b3e..0a4ba78e 100644 --- a/docs/output/targeted.md +++ b/docs/output/targeted.md @@ -250,13 +250,13 @@ This section contains the final output of the pipeline. It contains information - `cigar/` - `*_cutSite.json`: Contains the protospacer cut site position in the reference. - - `*_edition.html`: Interactive pie chart with the percentage of edition types. Reads are classified between WT (without an edit) and indels. Indels are divided between deletions, insertions and delins (deletion + insertion). Deletions and insertions can be out of frame or in frame. + - `*_edition.html`: Interactive pie chart with the percentage of edition types. Reads are classified between WT (without an edit) and indels. Indels are divided between deletions, insertions and delins (deletion + insertion). Deletions and insertions can be out of frame or in frame. A similar plot can be visualised in the MultiQC report. ![Test sample hCas9-AAVS1-a edition plot](../images/hCas9-AAVS1-a_edition.png) - `*_edits.csv`: Table containing the number of reads classified to each edition type. Contains the data visualized in the pie chart. - `*_indels.csv`: Table containing information of all reads. Edit type, edit start and length, if the edition happens above the error rate, if it's located into the common edit window, the frequency, the percentage, the pattern, surrounding nucleotides in case of insertions, the protospacer cut site, the sample id, number of aligned reads and number of reads with and without a template modification. - - `*_QC-indels.html`: Interactive pie chart with information about aligned reads. Reads are classified between WT and containing indels. Both types are classified between passing the filtering steps or not. Indel reads passing the filtering steps are divided in reads with a modification above the error rate and located in the common edit window, above the error rate but not in the edit region, vice versa, or any of those conditions. + - `*_QC-indels.html`: Interactive pie chart with information about aligned reads. Reads are classified between WT and containing indels. Both types are classified between passing the filtering steps or not. Indel reads passing the filtering steps are divided in reads with a modification above the error rate and located in the common edit window, above the error rate but not in the edit region, vice versa, or any of those conditions. A similar plot can be visualised in the MultiQC report. ![Test sample hCas9-AAVS1-a QC indels plot](../images/hCas9-AAVS1-a_QC-indels.png) - - `*_reads.html`: Interactive pie chart with percentage of the number of raw reads, reads merged with Pear, reads passing quality filters and UMI clustered reads. + - `*_reads.html`: Interactive pie chart with percentage of the number of raw reads, reads merged with Pear, reads passing quality filters and UMI clustered reads. A table with this information can be visualised in the MultiQC report. ![Test sample hCas9-AAVS1-a reads plot](../images/hCas9-AAVS1-a_reads.png) - `*_subs-perc.csv`: Table containing the percentage of each nucleotide found for each reference position. @@ -278,6 +278,17 @@ This section contains the final output of the pipeline. It contains information Results generated by MultiQC collate pipeline QC from supported tools e.g. FastQC. The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see . +`multiqc_report.html` contains statistics for FastQC and Cutadapt modules. It also contains a table with statistics about read processing (equivalent to `/cigar/*_reads.html` plots), and plots summarising the found editions (equivalent to `/cigar/*_edition.html` plots) and indel quality filters (equivalent to `/cigar/*_QC-indels.html` plots). + +
      +Custom sections example + +![Read processing table](../images/mqc_read_processing_summary.png) +![Type of edition plot](../images/mqc_type_of_edition.png) +![QC of indels plot](../images/mqc_qc_of_indels.png) + +
      + ## Pipeline information
      From 015097c25527ce18f50a7d49e8f942dfc2d69a98 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 3 Aug 2023 14:36:11 +0200 Subject: [PATCH 039/133] summary processes don't modify input file, fixing resume --- CHANGELOG.md | 2 ++ modules/local/alignment_summary.nf | 2 +- modules/local/clustering_summary.nf | 2 +- modules/local/merging_summary.nf | 2 +- templates/alignment_summary.py | 3 ++- templates/clustering_summary.py | 3 ++- templates/merging_summary.py | 2 +- workflows/crisprseq_targeted.nf | 9 +++++++++ 8 files changed, 19 insertions(+), 6 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index e5d26623..76d9871a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -12,6 +12,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- Summary processes don't modify the input file anymore, allowing resuming these processes ([#65](https://github.com/nf-core/crisprseq/pull/65)) + ### Deprecated ## [v2.0.0 - Paprika Lovelace](https://github.com/nf-core/crisprseq/releases/tag/2.0.0) - [05.07.2023] diff --git a/modules/local/alignment_summary.nf b/modules/local/alignment_summary.nf index 2805cd92..3520689f 100644 --- a/modules/local/alignment_summary.nf +++ b/modules/local/alignment_summary.nf @@ -11,7 +11,7 @@ process ALIGNMENT_SUMMARY { tuple val(meta), path(reads), path(summary) output: - tuple val(meta), path(summary), emit: summary + tuple val(meta), path("*_alignment_summary.csv"), emit: summary when: task.ext.when == null || task.ext.when diff --git a/modules/local/clustering_summary.nf b/modules/local/clustering_summary.nf index a134bdaf..b31d4f36 100644 --- a/modules/local/clustering_summary.nf +++ b/modules/local/clustering_summary.nf @@ -11,7 +11,7 @@ process CLUSTERING_SUMMARY { tuple val(meta), path(reads), path(summary) output: - tuple val(meta), path(summary), emit: summary + tuple val(meta), path("*_clustering_summary.csv"), emit: summary when: task.ext.when == null || task.ext.when diff --git a/modules/local/merging_summary.nf b/modules/local/merging_summary.nf index dcd31e04..cf9c158b 100644 --- a/modules/local/merging_summary.nf +++ b/modules/local/merging_summary.nf @@ -12,7 +12,7 @@ process MERGING_SUMMARY { output: - tuple val(meta), path("*_summary.csv"), emit: summary + tuple val(meta), path("*_merging_summary.csv"), emit: summary when: task.ext.when == null || task.ext.when diff --git a/templates/alignment_summary.py b/templates/alignment_summary.py index 5adedf93..12dd6109 100644 --- a/templates/alignment_summary.py +++ b/templates/alignment_summary.py @@ -12,7 +12,8 @@ summary_lines = summary.readlines() add_line = True -with open("$summary", "w") as output_file: +outname = "$summary".replace("_clustering_summary.csv", "_alignment_summary.csv") +with open(outname, "w") as output_file: for line in summary_lines: if "aligned-reads" not in line: output_file.write(line) diff --git a/templates/clustering_summary.py b/templates/clustering_summary.py index 208b089b..ea501964 100644 --- a/templates/clustering_summary.py +++ b/templates/clustering_summary.py @@ -12,7 +12,8 @@ summary_lines = summary.readlines() add_line = True -with open("$summary", "w") as output_file: +outname = "$summary".replace("_merging_summary.csv", "_clustering_summary.csv") +with open(outname, "w") as output_file: for line in summary_lines: if "clustered-reads" not in line: output_file.write(line) diff --git a/templates/merging_summary.py b/templates/merging_summary.py index 804a5be3..2d88e24a 100755 --- a/templates/merging_summary.py +++ b/templates/merging_summary.py @@ -34,7 +34,7 @@ else: prefix = "$meta.id" -with open(f"{prefix}_summary.csv", "w") as output_file: +with open(f"{prefix}_merging_summary.csv", "w") as output_file: output_file.write("class, count\\n") output_file.write(f"raw-reads, {raw_reads_count} (100.0%)\\n") output_file.write( diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 6556d79d..90917137 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -341,6 +341,15 @@ workflow CRISPRSEQ_TARGETED { .join(PEAR.out.assembled, remainder: true) .join(SEQTK_SEQ_MASK.out.fastx) .join(CUTADAPT.out.log) + .map { meta, reads, assembled, masked, trimmed -> + if (assembled == null) { + assembled = file('null_a') + } + if (trimmed == "null") { + trimmed = file('null_t') + } + return [ meta, reads, assembled, masked, trimmed ] + } .set { ch_merging_summary_data } } else { ch_cat_fastq.paired From 1d111f9fce66bc3dfb678689f4f36beca02adb4c Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 3 Aug 2023 14:56:46 +0200 Subject: [PATCH 040/133] don't stash unexistent files, use empty lists instead, fix AWS tests --- CHANGELOG.md | 2 ++ templates/merging_summary.py | 6 +++--- workflows/crisprseq_targeted.nf | 22 ++++++++++++---------- 3 files changed, 17 insertions(+), 13 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index e5d26623..e1ffeea7 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -12,6 +12,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed +- Do not stash unexistent files, use empty lists instead. Fixes AWS tests ([#67](https://github.com/nf-core/crisprseq/pull/67)) + ### Deprecated ## [v2.0.0 - Paprika Lovelace](https://github.com/nf-core/crisprseq/releases/tag/2.0.0) - [05.07.2023] diff --git a/templates/merging_summary.py b/templates/merging_summary.py index 804a5be3..ffc7d277 100755 --- a/templates/merging_summary.py +++ b/templates/merging_summary.py @@ -7,7 +7,7 @@ with gzip.open("${raw_reads[0]}", "rt") as handle: raw_reads_count = len(list(SeqIO.parse(handle, "fastq"))) -if "$assembled_reads" == "null_a": +if "$assembled_reads" == "": assembled_reads_count = 0 else: with gzip.open("$assembled_reads", "rt") as handle: @@ -16,7 +16,7 @@ with gzip.open("$trimmed_reads", "rt") as handle: trimmed_reads_count = len(list(SeqIO.parse(handle, "fastq"))) # Filtered reads -if "$trimmed_adapters" == "null_t": +if "$trimmed_adapters" == "": adapters_count = 0 adapters_percentage = "(0.0%)" else: @@ -41,7 +41,7 @@ f"merged-reads, {assembled_reads_count} ({round(assembled_reads_count * 100 / raw_reads_count,1)}%)\\n" ) output_file.write(f"reads-with-adapters, {adapters_count} {adapters_percentage}\\n") - if "$assembled_reads" == "null_a": + if "$assembled_reads" == "": output_file.write( f"quality-filtered-reads, {trimmed_reads_count} ({round(trimmed_reads_count * 100 / raw_reads_count,1)}%)\\n" ) diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 6556d79d..ebc07552 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -341,21 +341,23 @@ workflow CRISPRSEQ_TARGETED { .join(PEAR.out.assembled, remainder: true) .join(SEQTK_SEQ_MASK.out.fastx) .join(CUTADAPT.out.log) + .map { meta, reads, assembled, masked, trimmed -> + if (assembled == null) { + assembled = [] + } + return [ meta, reads, assembled, masked, trimmed ] + } .set { ch_merging_summary_data } } else { ch_cat_fastq.paired .mix(ch_cat_fastq.single) .join(PEAR.out.assembled, remainder: true) .join(SEQTK_SEQ_MASK.out.fastx) - .combine(Channel.value("null")) - .map { meta, reads, assembled, masked, trimmed -> + .map { meta, reads, assembled, masked -> if (assembled == null) { - assembled = file('null_a') + assembled = [] } - if (trimmed == "null") { - trimmed = file('null_t') - } - return [ meta, reads, assembled, masked, trimmed ] + return [ meta, reads, assembled, masked, [] ] } .set { ch_merging_summary_data } } @@ -705,13 +707,13 @@ workflow CRISPRSEQ_TARGETED { .join(ALIGNMENT_SUMMARY.out.summary) .map { meta, reads, index, reference, protospacer, template, template_bam, reference_template, summary -> if (template == null) { - template = file('null_t') + template = [] } if (template_bam == null) { - template_bam = file('null_b') + template_bam = [] } if (reference_template == null) { - reference_template = file('null_r') + reference_template = [] } return [meta, reads, index, reference, protospacer, template, template_bam, reference_template, summary] } From 7463aadcf562fdb0f33bfda55d009a5c8fc070f3 Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C3=BAlia=20Mir=20Pedrol?= Date: Thu, 3 Aug 2023 14:59:12 +0200 Subject: [PATCH 041/133] Update CHANGELOG.md --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 76d9871a..573ec572 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -12,7 +12,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed -- Summary processes don't modify the input file anymore, allowing resuming these processes ([#65](https://github.com/nf-core/crisprseq/pull/65)) +- Summary processes don't modify the input file anymore, allowing resuming these processes ([#66](https://github.com/nf-core/crisprseq/pull/66)) ### Deprecated From 354e1242a2eb66088b4ae1f23efcefcbb3664566 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Fri, 4 Aug 2023 13:49:03 +0200 Subject: [PATCH 042/133] rename merging_summary to preprocessing_summary --- conf/modules.config | 2 +- ...ng_summary.nf => preprocessing_summary.nf} | 6 ++--- templates/clustering_summary.py | 2 +- ...ng_summary.py => preprocessing_summary.py} | 2 +- workflows/crisprseq_targeted.nf | 26 +++++++++---------- 5 files changed, 19 insertions(+), 19 deletions(-) rename modules/local/{merging_summary.nf => preprocessing_summary.nf} (77%) rename templates/{merging_summary.py => preprocessing_summary.py} (96%) diff --git a/conf/modules.config b/conf/modules.config index 75765bcd..f78266e9 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -128,7 +128,7 @@ process { ext.prefix = { "${fasta.baseName}_top" } } - withName: MERGING_SUMMARY { + withName: PREPROCESSING_SUMMARY { publishDir = [ path: { "${params.outdir}/summary/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, mode: params.publish_dir_mode, diff --git a/modules/local/merging_summary.nf b/modules/local/preprocessing_summary.nf similarity index 77% rename from modules/local/merging_summary.nf rename to modules/local/preprocessing_summary.nf index cf9c158b..52d29e17 100644 --- a/modules/local/merging_summary.nf +++ b/modules/local/preprocessing_summary.nf @@ -1,4 +1,4 @@ -process MERGING_SUMMARY { +process PREPROCESSING_SUMMARY { tag "$meta.id" label 'process_single' @@ -12,11 +12,11 @@ process MERGING_SUMMARY { output: - tuple val(meta), path("*_merging_summary.csv"), emit: summary + tuple val(meta), path("*_preprocessing_summary.csv"), emit: summary when: task.ext.when == null || task.ext.when script: - template 'merging_summary.py' + template 'preprocessing_summary.py' } diff --git a/templates/clustering_summary.py b/templates/clustering_summary.py index ea501964..5b1dfd42 100644 --- a/templates/clustering_summary.py +++ b/templates/clustering_summary.py @@ -12,7 +12,7 @@ summary_lines = summary.readlines() add_line = True -outname = "$summary".replace("_merging_summary.csv", "_clustering_summary.csv") +outname = "$summary".replace("_preprocessing_summary.csv", "_clustering_summary.csv") with open(outname, "w") as output_file: for line in summary_lines: if "clustered-reads" not in line: diff --git a/templates/merging_summary.py b/templates/preprocessing_summary.py similarity index 96% rename from templates/merging_summary.py rename to templates/preprocessing_summary.py index f0e134e2..10cb94b1 100755 --- a/templates/merging_summary.py +++ b/templates/preprocessing_summary.py @@ -34,7 +34,7 @@ else: prefix = "$meta.id" -with open(f"{prefix}_merging_summary.csv", "w") as output_file: +with open(f"{prefix}_preprocessing_summary.csv", "w") as output_file: output_file.write("class, count\\n") output_file.write(f"raw-reads, {raw_reads_count} (100.0%)\\n") output_file.write( diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index ebc07552..56742701 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -38,14 +38,14 @@ ch_multiqc_custom_methods_description = params.multiqc_methods_description ? fil // // MODULE // -include { FIND_ADAPTERS } from '../modules/local/find_adapters' -include { EXTRACT_UMIS } from '../modules/local/extract_umis' -include { ORIENT_REFERENCE } from '../modules/local/orient_reference' -include { CIGAR_PARSER } from '../modules/local/cigar_parser' -include { MERGING_SUMMARY } from '../modules/local/merging_summary' -include { CLUSTERING_SUMMARY } from '../modules/local/clustering_summary' -include { ALIGNMENT_SUMMARY } from '../modules/local/alignment_summary' -include { TEMPLATE_REFERENCE } from '../modules/local/template_reference' +include { FIND_ADAPTERS } from '../modules/local/find_adapters' +include { EXTRACT_UMIS } from '../modules/local/extract_umis' +include { ORIENT_REFERENCE } from '../modules/local/orient_reference' +include { CIGAR_PARSER } from '../modules/local/cigar_parser' +include { PREPROCESSING_SUMMARY } from '../modules/local/preprocessing_summary' +include { CLUSTERING_SUMMARY } from '../modules/local/clustering_summary' +include { ALIGNMENT_SUMMARY } from '../modules/local/alignment_summary' +include { TEMPLATE_REFERENCE } from '../modules/local/template_reference' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -347,7 +347,7 @@ workflow CRISPRSEQ_TARGETED { } return [ meta, reads, assembled, masked, trimmed ] } - .set { ch_merging_summary_data } + .set { ch_preprocessing_summary_data } } else { ch_cat_fastq.paired .mix(ch_cat_fastq.single) @@ -359,14 +359,14 @@ workflow CRISPRSEQ_TARGETED { } return [ meta, reads, assembled, masked, [] ] } - .set { ch_merging_summary_data } + .set { ch_preprocessing_summary_data } } // // MODULE: Summary of merged reads // - MERGING_SUMMARY { - ch_merging_summary_data + PREPROCESSING_SUMMARY { + ch_preprocessing_summary_data } @@ -587,7 +587,7 @@ workflow CRISPRSEQ_TARGETED { // CLUSTERING_SUMMARY ( ch_preprocess_reads - .join(MERGING_SUMMARY.out.summary) + .join(PREPROCESSING_SUMMARY.out.summary) ) From 613ed313d77b1e3120fe718e81fefe8fab9a85b0 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Fri, 4 Aug 2023 13:50:05 +0200 Subject: [PATCH 043/133] update changelog --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 14bc880d..ac39b6f8 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -14,6 +14,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - Summary processes don't modify the input file anymore, allowing resuming these processes ([#66](https://github.com/nf-core/crisprseq/pull/66)) - Do not stash unexistent files, use empty lists instead. Fixes AWS tests ([#67](https://github.com/nf-core/crisprseq/pull/67)) +- Rename process `merging_summary` to `preprocessing_summary` to improve clarity ([#68](https://github.com/nf-core/crisprseq/pull/68)) ### Deprecated From 710911c1aed8ab9898aa4be6397aeb75370e535a Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C3=BAlia=20Mir=20Pedrol?= Date: Fri, 4 Aug 2023 13:52:42 +0200 Subject: [PATCH 044/133] Update CHANGELOG.md --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index ac39b6f8..1c24832a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -14,7 +14,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - Summary processes don't modify the input file anymore, allowing resuming these processes ([#66](https://github.com/nf-core/crisprseq/pull/66)) - Do not stash unexistent files, use empty lists instead. Fixes AWS tests ([#67](https://github.com/nf-core/crisprseq/pull/67)) -- Rename process `merging_summary` to `preprocessing_summary` to improve clarity ([#68](https://github.com/nf-core/crisprseq/pull/68)) +- Rename process `merging_summary` to `preprocessing_summary` to improve clarity ([#69](https://github.com/nf-core/crisprseq/pull/69)) ### Deprecated From 6ef05ac032b45ea40b451728d59738f582c89ee2 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 8 Aug 2023 09:07:13 +0200 Subject: [PATCH 045/133] Bagel2 --- conf/modules.config | 32 ++++++++++++++++++++++++++++++ modules/local/bagel2/bf.nf | 2 +- modules/local/bagel2/pr.nf | 2 +- modules/nf-core/mageck/mle/main.nf | 2 +- workflows/crisprseq_screening.nf | 21 ++++++-------------- 5 files changed, 41 insertions(+), 18 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 75765bcd..aee0939a 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -43,6 +43,38 @@ process { ] } + withName: BAGEL2_BF { + publishDir = [ + path: { "${params.outdir}/bagel2/bayes_factor/" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + + withName: BAGEL2_PR { + publishDir = [ + path: { "${params.outdir}/bagel2/precision_recall/" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + + withName: BAGEL2_FC { + publishDir = [ + path: { "${params.outdir}/bagel2/fold_change/" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + + withName: BAGEL2_GRAPH { + publishDir = [ + path: { "${params.outdir}/bagel2/graphs/" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] + } + withName: FASTQC { ext.args = '--quiet' publishDir = [ diff --git a/modules/local/bagel2/bf.nf b/modules/local/bagel2/bf.nf index 1353965c..f3a3060d 100644 --- a/modules/local/bagel2/bf.nf +++ b/modules/local/bagel2/bf.nf @@ -24,7 +24,7 @@ process BAGEL2_BF { def prefix = task.ext.prefix ?: "${meta.treatment}" """ - BAGEL.py bf -i $foldchange -o '${meta.treatment}.bf' $args -e $reference_essentials -n $reference_nonessentials -c ${meta.treatment} + BAGEL.py bf -i $foldchange -o '${meta.treatment}_vs_${meta.reference}.bf' $args -e $reference_essentials -n $reference_nonessentials -c ${meta.treatment} """ diff --git a/modules/local/bagel2/pr.nf b/modules/local/bagel2/pr.nf index 4a5616b4..b5cd1793 100644 --- a/modules/local/bagel2/pr.nf +++ b/modules/local/bagel2/pr.nf @@ -24,7 +24,7 @@ process BAGEL2_PR { def prefix = task.ext.prefix ?: "${meta.id}" """ - BAGEL.py pr -i $bf -o '${meta.treatment}.pr' -e $reference_essentials -n $reference_nonessentials + BAGEL.py pr -i $bf -o '${meta.treatment}_vs_${meta.reference}.pr' -e $reference_essentials -n $reference_nonessentials """ } diff --git a/modules/nf-core/mageck/mle/main.nf b/modules/nf-core/mageck/mle/main.nf index 626c410d..9c744afe 100644 --- a/modules/nf-core/mageck/mle/main.nf +++ b/modules/nf-core/mageck/mle/main.nf @@ -1,6 +1,6 @@ process MAGECK_MLE { tag "$meta.id" - label 'process_medium' + label 'process_high' conda "bioconda::mageck=0.5.9" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 317838d2..3682e4fe 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -101,6 +101,9 @@ workflow CRISPRSEQ_SCREENING { } } .set { ch_input } + + ch_value = Channel.value(single_end) + ch_value.dump(tag: "ch_value") // // MODULE: Run FastQC // @@ -125,7 +128,6 @@ workflow CRISPRSEQ_SCREENING { joined.dump(tag: "input joined") } - // // MODULE: Run mageck count // @@ -177,20 +179,9 @@ workflow CRISPRSEQ_SCREENING { .set { ch_bagel } counts = ch_bagel.combine(ch_counts) - //TO DO MAKE THIS PRETTIER - if(!params.bagel_reference_essentials) { - ch_bagel_reference_essentials = Channel.fromPath("${projectDir}/assets/CEGv2.txt") - } else { - ch_bagel_reference_essentials = Channel.fromPath(params.bagel_reference_essentials) - } - - //ch_bagel_reference_essentials.dump(tag: "input joined") - - //TO DO make this prettier - if(!params.bagel_reference_nonessentials) { - ch_bagel_reference_nonessentials = Channel.fromPath("${projectDir}/assets/NEGv1.txt") - } else { - ch_bagel_reference_nonessentials = Channel.fromPath(params.bagel_reference_nonessentials) } + //Define non essential and essential genes channels for bagel2 + ch_bagel_reference_essentials= params.bagel_reference_essentials ? Channel.fromPath(params.bagel_reference_essentials) : Channel.fromPath("${projectDir}/assets/CEGv2.txt") + ch_bagel_reference_nonessentials= params.bagel_reference_nonessentials ? Channel.fromPath(params.bagel_reference_nonessentials) : Channel.fromPath("${projectDir}/assets/NEGv1.txt") BAGEL2_FC ( From 7030145e69622588b6cc06bd132182abbe925211 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 8 Aug 2023 09:11:05 +0200 Subject: [PATCH 046/133] Add new features in changelog --- CHANGELOG.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index e5d26623..6284d434 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -9,9 +9,11 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - Template update v2.9 ([#52](https://github.com/nf-core/crisprseq/pull/52)) - Use `Channel.fromSamplesheet()` from `nf-validation` to validate input sample sheets and create an input channel ([#58](https://github.com/nf-core/crisprseq/pull/58)) +- BAGEL2 as a module which detects gene essentiality ### Fixed +- Change to `process_high` for the mageck mle module ### Deprecated ## [v2.0.0 - Paprika Lovelace](https://github.com/nf-core/crisprseq/releases/tag/2.0.0) - [05.07.2023] From 5be5815247eec5f0f29d8e965e5f96f0d03b70eb Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 8 Aug 2023 14:44:24 +0200 Subject: [PATCH 047/133] Fixed paired-end --- modules/nf-core/mageck/count/main.nf | 14 +++++++---- workflows/crisprseq_screening.nf | 36 +++++++++++++--------------- 2 files changed, 27 insertions(+), 23 deletions(-) diff --git a/modules/nf-core/mageck/count/main.nf b/modules/nf-core/mageck/count/main.nf index 01691029..ced51e85 100644 --- a/modules/nf-core/mageck/count/main.nf +++ b/modules/nf-core/mageck/count/main.nf @@ -1,6 +1,6 @@ process MAGECK_COUNT { tag "$meta.id" - label 'process_medium' + label 'process_high' conda "bioconda::mageck=0.5.9" container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? @@ -26,9 +26,15 @@ process MAGECK_COUNT { script: def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" - def input_file = ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz")) ? "--fastq ${inputfile}" : "-k ${inputfile}" + // def input_file = ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz")) ? "--fastq ${inputfile}" : "-k ${inputfile}" def sample_label = ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz")) ? "--sample-label ${meta.id}" : '' - + + if (meta.single_end && ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz"))) { + input = "--fastq ${inputfile}" + } else { + input = "--fastq ${inputfile[0]} --fastq-2 ${inputfile[1]}" + } + """ mageck \\ count \\ @@ -36,7 +42,7 @@ process MAGECK_COUNT { -l $library \\ -n $prefix \\ $sample_label \\ - $input_file \\ + $input cat <<-END_VERSIONS > versions.yml diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 3682e4fe..52b24c9a 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -88,45 +88,43 @@ workflow CRISPRSEQ_SCREENING { // // Create input channel from input file provided through params.input // - single_end = true Channel.fromSamplesheet("input") .map{ meta, fastq_1, fastq_2, x, y, z -> // x (reference), y (protospacer), and z (template) are part of the targeted workflows and we don't need them - if (!fastq_2) { - single_end = true - return [ meta, [ fastq_1 ] ] - } else { - single_end = false - return [ meta, [ fastq_1, fastq_2 ] ] - } + return [ meta + [ single_end:fastq_2?false:true ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] } .set { ch_input } - ch_value = Channel.value(single_end) - ch_value.dump(tag: "ch_value") + + ch_input.dump(tag: "ch_input") // // MODULE: Run FastQC // FASTQC ( ch_input ) + + ch_versions = ch_versions.mix(FASTQC.out.versions.first()) - if(single_end==true) { + ch_input - .map { meta, fastq -> - [meta.condition, fastq] + .map { meta, fastq -> + [meta.condition, fastq, meta.single_end] } .reduce { a, b -> - ["${a[0]},${b[0]}", a[1] + b[1]] + if(a[2] != b[2] ) { + error "Your samplesheet contains a mix of single-end and paired-end data. This is not supported." + } + return ["${a[0]},${b[0]}", a[1] + b[1], b[2]] } - .map { condition, fastqs -> - [[id: condition], fastqs] + .map { condition, fastqs, single_end -> + [[id: condition, single_end: single_end], fastqs] } .set { joined } joined.dump(tag: "input joined") - } + // // MODULE: Run mageck count @@ -187,7 +185,7 @@ workflow CRISPRSEQ_SCREENING { BAGEL2_FC ( counts ) - } + ch_bagel_bf = BAGEL2_FC.out.foldchange.combine(ch_bagel_reference_essentials) .combine(ch_bagel_reference_nonessentials) @@ -208,7 +206,7 @@ workflow CRISPRSEQ_SCREENING { BAGEL2_GRAPH ( BAGEL2_PR.out.pr ) - + } if(params.mle_design_matrix) { ch_mle = ch_counts.combine(ch_design) ch_mle.map { From 2519663efcfc77d5ebc74d5b6b27bfd7dd861a4c Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 9 Aug 2023 09:22:16 +0200 Subject: [PATCH 048/133] Update CHANGELOG.md MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index f086f1c3..f9f6159b 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -9,7 +9,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - Template update v2.9 ([#52](https://github.com/nf-core/crisprseq/pull/52)) - Use `Channel.fromSamplesheet()` from `nf-validation` to validate input sample sheets and create an input channel ([#58](https://github.com/nf-core/crisprseq/pull/58)) -- BAGEL2 as a module which detects gene essentiality +- BAGEL2 as a module which detects gene essentiality ([#60](https://github.com/nf-core/crisprseq/pull/60)) ### Fixed From 65c0f6f010619b4472d2df4e2d723d46743fb6ba Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 9 Aug 2023 09:22:35 +0200 Subject: [PATCH 049/133] Update CHANGELOG.md MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index f9f6159b..a3e18bae 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -13,7 +13,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed -- Change to `process_high` for the mageck mle module +- Change to `process_high` for the mageck mle module ([#60](https://github.com/nf-core/crisprseq/pull/60) - Summary processes don't modify the input file anymore, allowing resuming these processes ([#66](https://github.com/nf-core/crisprseq/pull/66)) - Do not stash unexistent files, use empty lists instead. Fixes AWS tests ([#67](https://github.com/nf-core/crisprseq/pull/67)) - Rename process `merging_summary` to `preprocessing_summary` to improve clarity ([#69](https://github.com/nf-core/crisprseq/pull/69)) From 2be7f3b4c9bf0d5a4079b707eb20e23bbec6b32d Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 9 Aug 2023 11:40:30 +0200 Subject: [PATCH 050/133] Update workflows/crisprseq_screening.nf MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- workflows/crisprseq_screening.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 52b24c9a..730478fe 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -69,7 +69,7 @@ include { CRISPRCLEANR_NORMALIZE } from '../modules/nf-core/crisprcleanr/no include { BAGEL2_FC } from '../modules/local/bagel2/fc' include { BAGEL2_BF } from '../modules/local/bagel2/bf' include { BAGEL2_PR } from '../modules/local/bagel2/pr' -include { BAGEL2_GRAPH } from '../modules/local/bagel2/graph' +include { BAGEL2_GRAPH } from '../modules/local/bagel2/graph' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ From ea07b75991267a372690d45927263891a91e8fa2 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 9 Aug 2023 11:41:04 +0200 Subject: [PATCH 051/133] Update workflows/crisprseq_screening.nf MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- workflows/crisprseq_screening.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 730478fe..dee43eeb 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -91,7 +91,7 @@ workflow CRISPRSEQ_SCREENING { Channel.fromSamplesheet("input") .map{ meta, fastq_1, fastq_2, x, y, z -> // x (reference), y (protospacer), and z (template) are part of the targeted workflows and we don't need them - return [ meta + [ single_end:fastq_2?false:true ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] + return [ meta + [ single_end:fastq_2?false:true ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] } .set { ch_input } From 9278293a2bfae6be856a45c11033c0fe068fb5db Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 9 Aug 2023 12:40:04 +0200 Subject: [PATCH 052/133] Update workflows/crisprseq_screening.nf MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- workflows/crisprseq_screening.nf | 2 -- 1 file changed, 2 deletions(-) diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index dee43eeb..94fbd7ee 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -95,8 +95,6 @@ workflow CRISPRSEQ_SCREENING { } .set { ch_input } - - ch_input.dump(tag: "ch_input") // // MODULE: Run FastQC // From 4cc4bba46cf578e75caa5de0588c80c515bed62c Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 9 Aug 2023 12:41:25 +0200 Subject: [PATCH 053/133] Update workflows/crisprseq_screening.nf MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- workflows/crisprseq_screening.nf | 2 -- 1 file changed, 2 deletions(-) diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 94fbd7ee..a9c3348a 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -121,8 +121,6 @@ workflow CRISPRSEQ_SCREENING { } .set { joined } - joined.dump(tag: "input joined") - // // MODULE: Run mageck count From d0b21d9d761812f4ef2dd218ff0840a3cd886b35 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 9 Aug 2023 12:43:09 +0200 Subject: [PATCH 054/133] formatting --- modules/local/bagel2/bf.nf | 4 +++- nextflow.config | 2 +- nextflow_schema.json | 4 ++-- workflows/crisprseq_screening.nf | 16 ++++++---------- 4 files changed, 12 insertions(+), 14 deletions(-) diff --git a/modules/local/bagel2/bf.nf b/modules/local/bagel2/bf.nf index f3a3060d..9f5fe090 100644 --- a/modules/local/bagel2/bf.nf +++ b/modules/local/bagel2/bf.nf @@ -10,7 +10,9 @@ process BAGEL2_BF { input: - tuple val(meta), path(foldchange), path(reference_essentials), path(reference_nonessentials) + tuple val(meta), path(foldchange) + path(reference_essentials) + path(reference_nonessentials) output: tuple val(meta), path("*.bf"), emit: bf diff --git a/nextflow.config b/nextflow.config index bf675e5f..eeeca587 100644 --- a/nextflow.config +++ b/nextflow.config @@ -21,7 +21,7 @@ params { count_table = null min_reads = 30 min_targeted_genes = 3 - bagel_reference_essentials = 'https://raw.githubusercontent.com/hart-lab/bagel/master/CEGv2.txt' + bagel_reference_essentials = 'https://raw.githubusercontent.com/hart-lab/bagel/master/CEGv2.txt' bagel_reference_nonessentials = 'https://raw.githubusercontent.com/hart-lab/bagel/master/NEGv1.txt' // Pipeline steps diff --git a/nextflow_schema.json b/nextflow_schema.json index c5774253..211f5f1a 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -196,12 +196,12 @@ "bagel_reference_essentials": { "type": "string", "description": "Core essential gene set for BAGEL2", - "default": "https://github.com/hart-lab/bagel/blob/master/CEGv2.txt" + "default": "https://raw.githubusercontent.com/hart-lab/bagel/master/CEGv2.txt" }, "bagel_reference_nonessentials": { "type": "string", "description": "Non essential gene set for BAGEL2", - "default": "https://github.com/hart-lab/bagel/blob/master/NEGv1.txt" + "default": "https://raw.githubusercontent.com/hart-lab/bagel/master/NEGv1.txt" } } }, diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 52b24c9a..586b8dc8 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -69,7 +69,7 @@ include { CRISPRCLEANR_NORMALIZE } from '../modules/nf-core/crisprcleanr/no include { BAGEL2_FC } from '../modules/local/bagel2/fc' include { BAGEL2_BF } from '../modules/local/bagel2/bf' include { BAGEL2_PR } from '../modules/local/bagel2/pr' -include { BAGEL2_GRAPH } from '../modules/local/bagel2/graph' +include { BAGEL2_GRAPH } from '../modules/local/bagel2/graph' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -178,24 +178,20 @@ workflow CRISPRSEQ_SCREENING { counts = ch_bagel.combine(ch_counts) //Define non essential and essential genes channels for bagel2 - ch_bagel_reference_essentials= params.bagel_reference_essentials ? Channel.fromPath(params.bagel_reference_essentials) : Channel.fromPath("${projectDir}/assets/CEGv2.txt") - ch_bagel_reference_nonessentials= params.bagel_reference_nonessentials ? Channel.fromPath(params.bagel_reference_nonessentials) : Channel.fromPath("${projectDir}/assets/NEGv1.txt") + ch_bagel_reference_essentials= params.bagel_reference_essentials ? Channel.value(params.bagel_reference_essentials) : Channel.value("${projectDir}/assets/CEGv2.txt") + ch_bagel_reference_nonessentials= params.bagel_reference_nonessentials ? Channel.value(params.bagel_reference_nonessentials) : Channel.value("${projectDir}/assets/NEGv1.txt") BAGEL2_FC ( counts ) - - ch_bagel_bf = BAGEL2_FC.out.foldchange.combine(ch_bagel_reference_essentials) - .combine(ch_bagel_reference_nonessentials) - - BAGEL2_BF ( - ch_bagel_bf + BAGEL2_FC.out.foldchange, + ch_bagel_reference_essentials, + ch_bagel_reference_nonessentials ) - BAGEL2_BF.out.bf.dump(tag:"BAGEL2_BF dump") ch_bagel_pr = BAGEL2_BF.out.bf.combine(ch_bagel_reference_essentials) .combine(ch_bagel_reference_nonessentials) From 6c8993bda1e6f7f3c1b71eeb996901e192ebaff1 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 9 Aug 2023 12:55:45 +0200 Subject: [PATCH 055/133] test --- modules/local/bagel2/graph.nf | 19 +------------------ 1 file changed, 1 insertion(+), 18 deletions(-) diff --git a/modules/local/bagel2/graph.nf b/modules/local/bagel2/graph.nf index 1bc149a4..4a48b7a3 100644 --- a/modules/local/bagel2/graph.nf +++ b/modules/local/bagel2/graph.nf @@ -1,20 +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 BAGEL2_GRAPH { tag "$meta.id" label 'process_single' @@ -29,9 +12,9 @@ process BAGEL2_GRAPH { tuple val(meta), path(pr) output: - //path "versions.yml" , emit: versions path("*.png") , emit: pictures + when: task.ext.when == null || task.ext.when From 29dc2bd6223e0daa1187b9ea419fa525d5349de4 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 9 Aug 2023 14:35:08 +0200 Subject: [PATCH 056/133] Update workflows/crisprseq_screening.nf MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- workflows/crisprseq_screening.nf | 1 - 1 file changed, 1 deletion(-) diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 9e020e1a..ff495e3a 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -194,7 +194,6 @@ workflow CRISPRSEQ_SCREENING { BAGEL2_PR ( ch_bagel_pr ) - BAGEL2_PR.out.pr.dump(tag:"TEST PR DUMP") BAGEL2_GRAPH ( BAGEL2_PR.out.pr ) From bc75826415e90d2d328cf682091f1919b6eb35c6 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 14 Aug 2023 15:45:15 +0200 Subject: [PATCH 057/133] don't save reference oriented files and other intermediate output files --- conf/modules.config | 16 ++++++++++++++-- docs/output/targeted.md | 4 +--- 2 files changed, 15 insertions(+), 5 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index f78266e9..867f0d58 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -23,7 +23,8 @@ process { publishDir = [ path: { "${params.outdir}/preprocessing/sequences" }, mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + saveAs: { filename -> filename.equals('versions.yml') ? null : filename }, + enabled: false ] } @@ -132,7 +133,8 @@ process { publishDir = [ path: { "${params.outdir}/summary/${task.process.tokenize(':')[-1].tokenize('_')[0].toLowerCase()}" }, mode: params.publish_dir_mode, - saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + saveAs: { filename -> filename.equals('versions.yml') ? null : filename }, + enabled: false ] } @@ -159,11 +161,21 @@ process { withName: RACON_1 { ext.args = '-t 4 -m 8 -x -6 -g -8 -w 500 --no-trimming' ext.prefix = { "${reads.baseName}_cycle1" } + publishDir = [ + path: { "${params.outdir}/racon_umi" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] } withName: RACON_2 { ext.args = '-t 4 -m 8 -x -6 -g -8 -w 500 --no-trimming' ext.prefix = { "${reads.baseName}_cycle2" } + publishDir = [ + path: { "${params.outdir}/racon_umi" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] } withName: MEDAKA { diff --git a/docs/output/targeted.md b/docs/output/targeted.md index 0a4ba78e..66f3d8fd 100644 --- a/docs/output/targeted.md +++ b/docs/output/targeted.md @@ -44,9 +44,7 @@ The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes d Output files - `preprocessing/sequences/` - - `*_reference.fasta`: Sequence used as a reference. - `*_template.fasta`: Provided template sequence. - - `*_correctOrient.fasta`: Reference sequence in the correct orientation. - `_NewReference.fasta`: New reference generated from adding the changes made by the template to the original reference. - `*_template-align.bam`: Alignment of the new reference (with template changes) to the original reference. @@ -174,7 +172,7 @@ The FastQC plots displayed in the MultiQC report shows _untrimmed_ reads. They m [Minimap2](https://github.com/lh3/minimap2) is a sequence alignment program that aligns DNA sequences against a reference database. -### racon +### racon_umi
      Output files From f8210a462f328ff41176873780d769b5550f87aa Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 16 Aug 2023 14:41:02 +0200 Subject: [PATCH 058/133] Add doc --- CITATIONS.md | 4 ++++ docs/output/screening.md | 20 ++++++++++++++++++++ workflows/crisprseq_screening.nf | 11 +++-------- 3 files changed, 27 insertions(+), 8 deletions(-) diff --git a/CITATIONS.md b/CITATIONS.md index e279aae8..2f5384ed 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -63,6 +63,10 @@ > Li, W. et al. Quality control, modeling, and visualization of CRISPR screens with MAGeCK-VISPR. Genome Biology 16, 281, doi:10.1186/s13059-015-0843-6 (2015). +- [BAGEL2](https://pubmed.ncbi.nlm.nih.gov/33407829/) + + > Kim E, Hart T. Improved analysis of CRISPR fitness screens and reduced off-target effects with the BAGEL2 gene essentiality classifier. Genome Med. 2021 Jan 6;13(1):2. doi: 10.1186/s13073-020-00809-3. PMID: 33407829; PMCID: PMC7789424. + - [BioContainers](https://pubmed.ncbi.nlm.nih.gov/28379341/) > da Veiga Leprevost F, Grüning B, Aflitos SA, Röst HL, Uszkoreit J, Barsnes H, Vaudel M, Moreno P, Gatto L, Weber J, Bai M, Jimenez RC, Sachsenberg T, Pfeuffer J, Alvarez RV, Griss J, Nesvizhskii AI, Perez-Riverol Y. BioContainers: an open-source and community-driven framework for software standardization. Bioinformatics. 2017 Aug 15;33(16):2580-2582. doi: 10.1093/bioinformatics/btx192. PubMed PMID: 28379341; PubMed Central PMCID: PMC5870671. diff --git a/docs/output/screening.md b/docs/output/screening.md index bb45fdcc..e51bb43a 100644 --- a/docs/output/screening.md +++ b/docs/output/screening.md @@ -97,6 +97,26 @@ The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes d [MAGeCK](https://sourceforge.net/p/mageck/wiki/Home/) is a computational tool to identify important genes from CRISPR-Cas9 screens. +### BAGEL2 + +
      +Output files + +- `bagel2/fold_change` + - `*.foldchange`: foldchange between the reference and treatment contrast provided +- `bagel2/bayes_factor` + - `*.bf`: bayes factor per gene +- `bagel2/precision_recall` + - `*.pr`: precision recall per gene +- `bagel2/graphs` + - `barplot*.png`: barplot of the bayes factor distribution + - `PR*.png`: precision recall plot (Recall vs FDR) + +
      + +[bagel2](https://github.com/hart-lab/bagel) is a computational tool to identify important essential genes for CRISPR-Cas9 screening experiments. + + ## MultiQC
      diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index ff495e3a..54a2d3b0 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -15,10 +15,6 @@ log.info logo + paramsSummaryLog(workflow) + citation WorkflowCrisprseq.initialise(params, log) -// Check input path parameters to see if they exist -def checkPathParamList = [ params.multiqc_config, params.reference_fasta, params.library] -for (param in checkPathParamList) { if (param) { file(param, checkIfExists: true) } } - // Check mandatory parameters //if (!params.count_table) { ch_input = file(params.input) } else { error('Input samplesheet not specified!') } if (params.library) { ch_library = file(params.library) } @@ -91,7 +87,7 @@ workflow CRISPRSEQ_SCREENING { Channel.fromSamplesheet("input") .map{ meta, fastq_1, fastq_2, x, y, z -> // x (reference), y (protospacer), and z (template) are part of the targeted workflows and we don't need them - return [ meta + [ single_end:fastq_2?false:true ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] + return [ meta + [ single_end:fastq_2?false:true ], fastq_2?[ fastq_1, fastq_2 ]:[ fastq_1 ] ] } .set { ch_input } @@ -174,9 +170,8 @@ workflow CRISPRSEQ_SCREENING { counts = ch_bagel.combine(ch_counts) //Define non essential and essential genes channels for bagel2 - ch_bagel_reference_essentials= params.bagel_reference_essentials ? Channel.value(params.bagel_reference_essentials) : Channel.value("${projectDir}/assets/CEGv2.txt") - ch_bagel_reference_nonessentials= params.bagel_reference_nonessentials ? Channel.value(params.bagel_reference_nonessentials) : Channel.value("${projectDir}/assets/NEGv1.txt") - + ch_bagel_reference_essentials= Channel.value(params.bagel_reference_essentials) + ch_bagel_reference_nonessentials= Channel.value(params.bagel_reference_nonessentials) BAGEL2_FC ( counts From e21fc25b08ee19f81f2fb304311859410f5b3519 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 16 Aug 2023 14:58:28 +0200 Subject: [PATCH 059/133] fix meta id --- modules/local/bagel2/graph.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/bagel2/graph.nf b/modules/local/bagel2/graph.nf index 4a48b7a3..8ca0d84c 100644 --- a/modules/local/bagel2/graph.nf +++ b/modules/local/bagel2/graph.nf @@ -1,5 +1,5 @@ process BAGEL2_GRAPH { - tag "$meta.id" + tag "${meta.treatment}_${meta.reference}" label 'process_single' conda "python=3.11.4 pandas=2.0.3 numpy=1.25.1 scikit-learn=1.3.0 click=8.1.6 matplotlib=3.7.2" From 63f328a42ffb85910bc30a5f6fa8afd7696ab5ff Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 16 Aug 2023 15:16:54 +0200 Subject: [PATCH 060/133] Ran nf-core modules patch --- .../nf-core/mageck/count/mageck-count.diff | 32 +++++++++++++++++-- 1 file changed, 29 insertions(+), 3 deletions(-) diff --git a/modules/nf-core/mageck/count/mageck-count.diff b/modules/nf-core/mageck/count/mageck-count.diff index 87f8e83c..53659a20 100644 --- a/modules/nf-core/mageck/count/mageck-count.diff +++ b/modules/nf-core/mageck/count/mageck-count.diff @@ -1,6 +1,14 @@ Changes in module 'nf-core/mageck/count' --- modules/nf-core/mageck/count/main.nf +++ modules/nf-core/mageck/count/main.nf +@@ -1,6 +1,6 @@ + process MAGECK_COUNT { + tag "$meta.id" +- label 'process_medium' ++ label 'process_high' + + conda "bioconda::mageck=0.5.9" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? @@ -12,8 +12,12 @@ path(library) @@ -15,14 +23,32 @@ Changes in module 'nf-core/mageck/count' path "versions.yml" , emit: versions when: -@@ -22,7 +26,7 @@ +@@ -22,9 +26,15 @@ script: def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" - def input_file = ("$inputfile".endsWith(".fastq.gz")) ? "--fastq ${inputfile}" : "-k ${inputfile}" -+ def input_file = ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz")) ? "--fastq ${inputfile}" : "-k ${inputfile}" ++ // def input_file = ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz")) ? "--fastq ${inputfile}" : "-k ${inputfile}" def sample_label = ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz")) ? "--sample-label ${meta.id}" : '' - +- ++ ++ if (meta.single_end && ("$inputfile".endsWith(".fastq.gz") || "$inputfile".endsWith(".fq.gz"))) { ++ input = "--fastq ${inputfile}" ++ } else { ++ input = "--fastq ${inputfile[0]} --fastq-2 ${inputfile[1]}" ++ } ++ """ + mageck \\ + count \\ +@@ -32,7 +42,7 @@ + -l $library \\ + -n $prefix \\ + $sample_label \\ +- $input_file \\ ++ $input + + + cat <<-END_VERSIONS > versions.yml ************************************************************ From 90eb2879b60f5b5752e86e4e79d040950558f8d3 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 16 Aug 2023 15:42:49 +0200 Subject: [PATCH 061/133] Add license --- assets/CEGv2.txt | 685 ---------------------------------- assets/NEGv1.txt | 929 ----------------------------------------------- bin/BAGEL.py | 52 ++- 3 files changed, 40 insertions(+), 1626 deletions(-) delete mode 100644 assets/CEGv2.txt delete mode 100644 assets/NEGv1.txt diff --git a/assets/CEGv2.txt b/assets/CEGv2.txt deleted file mode 100644 index fa3ce335..00000000 --- a/assets/CEGv2.txt +++ /dev/null @@ -1,685 +0,0 @@ -GENE HGNC_ID ENTREZ_ID -AARS HGNC:20 16 -ABCE1 HGNC:69 6059 -ABCF1 HGNC:70 23 -ACTB HGNC:132 60 -ACTL6A HGNC:24124 86 -ACTR10 HGNC:17372 55860 -ACTR2 HGNC:169 10097 -ADSL HGNC:291 158 -ADSS HGNC:292 159 -AHCY HGNC:343 191 -ALG1 HGNC:18294 56052 -ALG14 HGNC:28287 199857 -ALG2 HGNC:23159 85365 -ANAPC2 HGNC:19989 29882 -ANAPC4 HGNC:19990 29945 -ANAPC5 HGNC:15713 51433 -AQR HGNC:29513 9716 -ARCN1 HGNC:649 372 -ARIH1 HGNC:689 25820 -ARL2 HGNC:693 402 -ATP2A2 HGNC:812 488 -ATP5F1A HGNC:823 498 -ATP5F1B HGNC:830 506 -ATP5F1C HGNC:833 509 -ATP5F1D HGNC:837 513 -ATP5J2-PTCD1 NA -ATP5MG HGNC:14247 10632 -ATP5PO HGNC:850 539 -ATP6V0B HGNC:861 533 -ATP6V0C HGNC:855 527 -ATP6V1A HGNC:851 523 -ATP6V1D HGNC:13527 51382 -ATP6V1E1 HGNC:857 529 -ATR HGNC:882 545 -AURKB HGNC:11390 9212 -BANF1 HGNC:17397 8815 -BIRC5 HGNC:593 332 -BUB1B HGNC:1149 701 -BUB3 HGNC:1151 9184 -BUD31 HGNC:29629 8896 -BYSL HGNC:1157 705 -TWNK 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Traver Hart , Eiru Kim 2017. + +# Acknowledgements: John McGonigle +# modified 10/2019 +# Free to modify and redistribute with attribution +# --------------------------------- + +# ------------------------------------ +# constants + +""" +MIT License + +Copyright (c) 2020 Hart Lab + +Permission is hereby granted, free of charge, to any person obtaining a copy +of this software and associated documentation files (the "Software"), to deal +in the Software without restriction, including without limitation the rights +to use, copy, modify, merge, publish, distribute, sublicense, and/or sell +copies of the Software, and to permit persons to whom the Software is +furnished to do so, subject to the following conditions: + +The above copyright notice and this permission notice shall be included in all +copies or substantial portions of the Software. + +THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR +IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, +FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE +AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER +LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, +OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE +SOFTWARE. +""" + + + import click import numpy as np import pandas as pd @@ -54,17 +92,7 @@ def full_process_value(self, ctx, value): return value -# --------------------------------- -# BAGEL: Bayesian Analysis of Gene EssentaLity -# (c) Traver Hart , Eiru Kim 2017. -# Acknowledgements: John McGonigle -# modified 10/2019 -# Free to modify and redistribute with attribution -# --------------------------------- - -# ------------------------------------ -# constants @@ -822,7 +850,7 @@ def calculate_bayes_factors( multiple_factor = equalise_sgrna_no / float(len(gene2rna[g])) bf_norm[g] = targetbf[g] * multiple_factor - ''' + ''' if bf_std[rnatag] == 0.0: bf_norm[rnatag] = float('inf') else: @@ -934,7 +962,7 @@ def calculate_bayes_factors( fp.close() - + # # print out results From 54f65fe29a0ee106445922a5f8ff0eb3febe7d87 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Fri, 18 Aug 2023 15:10:19 +0200 Subject: [PATCH 062/133] add R script to generate more output plots (some modifications from original to fix bugs and make it run) --- bin/cigar_parser.R | 3 +- bin/plotter.R | 665 +++++++++++++++++++++++++++++ conf/modules.config | 4 + modules/local/cigar_parser.nf | 2 +- modules/local/crisprseq_plotter.nf | 42 ++ workflows/crisprseq_targeted.nf | 19 +- 6 files changed, 727 insertions(+), 8 deletions(-) create mode 100755 bin/plotter.R create mode 100644 modules/local/crisprseq_plotter.nf diff --git a/bin/cigar_parser.R b/bin/cigar_parser.R index 6bb972ca..2018d686 100755 --- a/bin/cigar_parser.R +++ b/bin/cigar_parser.R @@ -946,7 +946,8 @@ if (dim(alignment_info)[1] != 0){ dels <- separated_indels %>% filter(Modification == "del") dels_count <- dim(dels)[1] if ( t_type == "dels-out" || t_type == "dels-in"){ - dels_count <- dels_count - t_reads + t_in_dels <- dels %>% filter(Ids %in% t_ids[[1]]) + dels_count <- dels_count - dim(t_in_dels)[1] } ins <- separated_indels %>% filter(Modification == "ins") ins_count <- dim(ins)[1] diff --git a/bin/plotter.R b/bin/plotter.R new file mode 100755 index 00000000..10ab023d --- /dev/null +++ b/bin/plotter.R @@ -0,0 +1,665 @@ +#!/usr/bin/env Rscript + +############################ +#### Plot editing results +#### author: Marta Sanvicente +#### modified by: Júlia Mir @mirpedrol +#### Released under the MIT license. See git repository (https://github.com/nf-core/crisprseq) for full license text. +#### Original code https://bitbucket.org/synbiolab/crispr-a_nextflow/src/master/bin/get_plots.R +############################ + +args = commandArgs(trailingOnly=TRUE) + +library(ggplot2) +library(plyr) +library(dplyr) +library(seqinr) +library(ShortRead) +library(ggpubr) +library(ggmsa) +library(seqmagick) +library(stringr) +library(tidyr) +library(ggseqlogo) +library(plotly) +library(cowplot) +library(optparse) + +#################################### +### Load command line arguments #### +#################################### + +option_list = list( + make_option(c("-i", "--indels_info"), type="character", default=NULL, + help="CSV with information of INDEL editions", metavar="character"), + make_option(c("-r", "--reference"), type="character", default=NULL, + help="Reference fasta file", metavar="character"), + make_option(c("-g", "--gRNA_sequence"), type="character", default=NULL, + help="gRNA sequence", metavar="character"), + make_option(c("-n", "--sample_name"), type="character", default=NULL, + help="Sample ID", metavar="character"), + make_option(c("-c", "--cut_site"), type="numeric", default=NULL, + help="Cut position", metavar="numeric"), + make_option(c("-s", "--substitutions_info"), type="character", default=NULL, + help="CSV with information of substitution editions", metavar="character") +); + +opt_parser = OptionParser(option_list=option_list); +opt = parse_args(opt_parser); + +sample_name <- opt$sample_name +indels_info <- opt$indels_info +reference <- opt$reference +gR <- opt$gRNA_sequence +substitutions_info <- opt$substitutions_info +rel_cut_site <- as.numeric(opt$cut_site) + +data <- read.csv(indels_info) +ref_seq <- readFasta(reference) +subs_plup <- read.csv(substitutions_info, row.names = 1) + + +###################### +##### CRISPR-GA-1 like plot https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4184265/ +###################### + +plotIndels_gg <- function(indels_data, cut){ + # Split insertions from deletions + l <- length(indels_data$Modification) + indels_data$wt_reads[1] + data$t_reads[1] + dels <- indels_data %>% filter(Modification == "del") + ins <- indels_data %>% filter(Modification == "ins") + + #Metadata deletion + if (length(dels$X)>0){ + metadels <- as.data.frame(table(dels$Start,dels$Length)) + metadels <- metadels[which(metadels$Freq>0),] + colnames(metadels) <- c("Start","length","Freq") + metadels$End <- 0 + metadels$changes <- sum((as.integer(as.character(metadels$Freq)) * as.integer(as.character(metadels$length)))) #Now the changes column corrspond to a constant that will be used in the creation of accumulative. This variable stores the total number of changes per position + + for (i in 1:length(metadels$Start)){ + metadels$End[i] = as.integer(as.character(metadels$Start[i])) + as.integer(as.character(metadels$length[i])) - 1 + } + }else{ + metadels <- list() + } + + exportJson <- jsonlite::toJSON(metadels) + write(exportJson, paste(sample_name,"_metadels.json",sep="")) + + #Metadata insertion + if(length(ins$X)>0){ + metains <- as.data.frame(table(ins$Start,ins$Length)) + metains <- metains[which(metains$Freq>0),] + colnames(metains) <- c("Start","length","Freq") + metains$End <- 0 + metains$changes <- sum((as.integer(as.character(metains$Freq)) * as.integer(as.character(metains$length)))) #Now the changes column corrspond to a constant that will be used in the creation of accumulative. This variable stores the total number of changes per position + + for (i in 1:length(metains$Start)){ + metains$End[i] = as.integer(as.character(metains$Start[i])) + as.integer(as.character(metains$length[i])) - 1 + } + }else{ + metains <- list() + } + exportJson <- jsonlite::toJSON(metains) + write(exportJson, paste(sample_name,"_metains.json",sep="")) + + + # Count accumulated insertions and deletions by position + accum.dels=c() + accum.dels=unlist(apply(dels,1,function(X) return( seq(as.integer(X[3]), as.integer(X[3]) + as.integer(X[4])) ))) + accum.ins=c() + accum.ins=unlist(apply(ins,1,function(X) return( seq(as.integer(X[3]), as.integer(X[3]) + as.integer(X[4])) ))) + + # Plot + a_dels <- data.frame(location = c(accum.dels)) + a_ins <- data.frame(location = c(accum.ins)) + + + ############################################### + # ACUMMULATIVE LOCATIONS # + ############################################### + if ( dim(dels)[1] != 0 ){ + acc_dels <- as.data.frame(table(a_dels)) + acc_dels <- acc_dels[acc_dels$Freq>0,] + acc_dels$percent <- (acc_dels$Freq/sum(acc_dels$Freq))*100 + del_acumm <- plot_ly(y = ~acc_dels$percent, x= ~as.numeric(as.character(acc_dels$location)),type = "bar",hovertemplate = paste('Percentage: %{y}','Counts: ',acc_dels$Freq)) %>% + layout(xaxis = list(title="Accumulative Deletion Location",rangeslider = list()), yaxis = list(fixedrange = FALSE,title = "Reads Percentage(%)"), barmode = 'stack')%>% + layout(shapes = list(list( + type = "line", + y0 = 0, + y1 = 100, + yref = "paper", + x0 = cut_site, + x1 = cut_site, + line = list(color = "red", dash="dot") + )))%>% + add_text(showlegend = FALSE, x=cut_site, y=max(acc_dels$percent),text = "Cut site", hovertemplate = paste("Cut site ", cut_site), + textfont = list(color= c("red"))) + }else { + acc_dels <- data.frame() + } + + + if ( dim(ins)[1] != 0 ){ + acc_ins <- as.data.frame(table(a_ins)) + acc_ins <- acc_ins[acc_ins$Freq>0,] + acc_ins$percent <- (acc_ins$Freq/sum(acc_ins$Freq))*100 + + ins_acumm <- plot_ly(y = ~acc_ins$percent, x= ~as.numeric(as.character(acc_ins$location)),type = "bar",hovertemplate = paste('Percentage: %{y}','Counts: ',acc_ins$Freq)) %>% + layout(xaxis = list(title="Accumulative Insertion Location",rangeslider = list()), yaxis = list(fixedrange = FALSE,title = "Reads Percentage(%)"), barmode = 'stack')%>% + layout(shapes = list(list( + type = "line", + y0 = 0, + y1 = 100, + yref = "paper", + x0 = cut_site, + x1 = cut_site, + line = list(color = "red", dash="dot") + )))%>% + add_text(showlegend = FALSE, x=cut_site, y=max(acc_ins$percent),text = "Cut site", hovertemplate = paste("Cut Site ", cut_site), + textfont = list(color= c("red"))) + }else { + acc_ins <- data.frame() + } + + if( (dim(ins)[1] != 0) && (dim(dels)[1] != 0) ){ + accummulative <- subplot(del_acumm, ins_acumm, shareY = T,shareX = F,titleX = T,titleY = T) + } else if( dim(ins)[1] != 0 ){ + accummulative <- subplot(ins_acumm, shareY = T,shareX = F,titleX = T,titleY = T) + } else { + accummulative <- subplot(del_acumm, shareY = T,shareX = F,titleX = T,titleY = T) + } + + #Elements for dynamic table + sequece_length <- max(indels_data$Start+indels_data$Length) + exportJson <- jsonlite::toJSON(sequece_length) + write(exportJson, paste(sample_name,"_length.json",sep="")) + + exportJson <- jsonlite::toJSON(l) + write(exportJson, paste(sample_name,"_Total_reads.json",sep="")) + + + #INSERTIONS + ############################################### + # LOCATION # + ############################################### + + pattern = ins + + #defensive programming against file with no insertions + if(length(pattern$Modification)>0){ + + LOC <- as.data.frame(table(pattern$Start,pattern$Length,pattern$ins_nt)) + LOC <- LOC[which(LOC$Freq>0),] + LOC$percent <- (LOC$Freq/l)*100 + + + #AXIS -> PERCENTAGE, BAR -> COUNT + Ins_location <- plot_ly() %>% + layout(xaxis = list(title="Insertion Location",rangeslider = list()), yaxis = list(fixedrange = FALSE,title = "Reads Percentage(%)"), barmode = 'stack') + + Ins_location <- Ins_location %>% add_trace(x = ~as.numeric(as.character(LOC$Var1)), + y= ~LOC$percent, color = ~LOC$Var3, type = 'bar', + marker = list(~LOC$Freq), legendgroup = ~LOC$Var3, + hovertemplate = paste('Percentage: %{y}','Counts: ',LOC$Freq)) + + + Ins_location <- Ins_location %>% layout(shapes = list(list( + type = "line", + y0 = 0, + y1 = 100, + yref = "paper", + x0 = cut_site, + x1 = cut_site, + line = list(color = "red", dash="dot") + ))) %>% add_text(showlegend = FALSE, x=cut_site,y=max(LOC$percent),text = "Cut site", hovertemplate = paste("Cut site ", cut_site), + textfont = list(color= c("red"))) + + + ############################################### + # SIZES # + ############################################### + + #AXIS -> PERCENTAGE, BAR -> COUNT + Ins_sizes <- plot_ly() %>% + layout(xaxis = list(title="Insertion Sizes",rangeslider = list()), yaxis = list(title = "Reads Percentage(%)"), barmode = 'stack') + Ins_sizes <- Ins_sizes %>% add_trace(x = ~as.numeric(as.character(LOC$Var2)), + y= ~LOC$percent, color = ~LOC$Var3, type = 'bar', + marker = list(~LOC$Freq), showlegend = F, legendgroup = ~LOC$Var3, + hovertemplate = paste(' Percentage: %{y}','Count: ',LOC$Freq)) + + ############################################### + # MERGE PLOTS # + ############################################### + + insertions <- subplot(Ins_location,Ins_sizes, titleY = F, titleX = T) %>% layout(title = "Insertions",yaxis = list(title = "Reads Percentage (%)")) + }else{ + insertions<-empty_plot("No Insertions were detected.") + } + + #DELETIONS + #pattern filter (A,C,T,G) -> NHEJ + pattern <- dels + + #defensive programming against file with no insertions + if(length(pattern$Modification)){ + + for(i in 1:length(pattern$patterns)){ + if(length(strsplit(pattern$patterns[i],split = "")[[1]])==1){ + pattern$patterns[i] <- "NHEJ" + } + } + + ############################################### + # LOCATION # + ############################################### + + + LOC <- as.data.frame(table(pattern$Start,pattern$patterns,pattern$Length)) + LOC <- LOC[which(LOC$Freq>0),] + LOC$percent <- (LOC$Freq/l)*100 + + + #AXIS -> PERCENTAGE, BAR -> COUNT + Del_location <- plot_ly() %>% + layout(xaxis = list(title="Deletion Location",rangeslider = list()), yaxis = list(fixedrange = FALSE,title = "Reads Percentage(%)"), barmode = 'stack') + Del_location <- Del_location %>% add_trace(x = ~as.numeric(as.character(LOC$Var1)), + y= ~LOC$percent, color = ~LOC$Var2, type = 'bar', + marker = list(~LOC$Freq), legendgroup = ~LOC$Var2, + hovertemplate = paste('Percentage: %{y}','Counts: ',LOC$Freq)) + + Del_location <- Del_location %>% layout(shapes = list(list( + type = "line", + y0 = 0, + y1 = 100, + yref = "paper", + x0 = cut_site, + x1 = cut_site, + line = list(color = "red", dash="dot") + )))%>% + add_text(showlegend = FALSE, x=cut_site,y=max(LOC$percent),text = "Cut site", hovertemplate = paste("Cut site ", cut_site), + textfont = list(color= c("red"))) + + + ############################################### + # SIZES # + ############################################### + + #AXIS -> PERCENTAGE, BAR -> COUNT + Del_sizes <- plot_ly() %>% + layout(xaxis = list(title="Deletion Sizes",rangeslider = list()), yaxis = list(title = "Reads Percentage(%)"), barmode = 'stack') + Del_sizes <- Del_sizes %>% add_trace(x = ~as.numeric(as.character(LOC$Var3)), + y= ~LOC$percent, color = ~LOC$Var2, type = 'bar', + marker = list(~LOC$Freq), showlegend=F, legendgroup = ~LOC$Var2, + hovertemplate = paste('Size: ',LOC$Var3,' Percentage: %{y}')) + + + ############################################### + # MERGE PLOTS # + ############################################### + + deletions <- subplot(Del_location,Del_sizes, titleY = F, titleX = T) %>% layout(title = "Deletions",yaxis = list(title = "Reads Percentage (%)")) + }else{ + deletions<-empty_plot("No deletions were detected.") + } + plots <- list(deletions,insertions,accummulative) + return(plots) +} + +####### +#### Reverse complement +####### +strComp=function(X){ + return(c2s(rev(comp(s2c(X))))) +} + +###### +### Cut site +###### +get_cutSite <- function(gRNA_seq, reference, rel_cut){ + ### gRNA seq to upper case + gRNA_seq <- toupper(gRNA_seq) + ### Check orientation of gRNA in reference sequence and cut site + rvComp_seq <- strComp(as.character(sread(reference)[[1]])) + align <- pairwiseAlignment(toupper(gRNA_seq), toupper(as.character(sread(reference)[[1]])), type="global-local") + alignRevComp <- pairwiseAlignment(toupper(gRNA_seq), toupper(rvComp_seq), type="global-local") + + if (score(align) > score(alignRevComp)){ + if (rel_cut > 0){ + cut_site <- start(subject(align))+rel_cut-1 + } else { + cut_site <- start(subject(align)) + ( nchar(gRNA_seq) + rel_cut ) -1 + } + } else { + if (rel_cut > 0){ + cut_site <- nchar(as.character(sread(reference)[[1]])) - end(subject(alignRevComp)) + (nchar(gRNA_seq)-rel_cut) + } else { + cut_site <- nchar(as.character(sread(reference)[[1]])) - end(subject(alignRevComp)) - rel_cut + } + } + return(cut_site) +} + + +####### +#### Empty Plots +####### +empty_plot <- function(title = NULL){ + p <- plotly_empty(type = "scatter", mode = "markers") %>% + config( + displayModeBar = FALSE + ) %>% + layout( + title = list( + text = title, + yref = "paper", + y = 0.5 + ) + ) + return(p) +} + +###################### +##### Alleles percentages plot +###################### + +indel_count_seq <- function(indel_data){ + # Get table with percentages of indels among all genotypes + indel_count <- indel_data %>% group_by(Start, Length, Modification) %>% dplyr::summarise(freq=n()) + #indel_count <- plyr::count(indel_data, vars = c("Start", "Length", "Modification")) + indel_count <- indel_count[order(indel_count$freq, decreasing = TRUE),] + indel_count$Perc <- (indel_count$freq/sum(indel_count$freq)) * 100 + return(as.data.frame(indel_count)) +} + +get_sequences <- function(indels, ref_seq){ + edited_sequences <- c() + seqs <- lapply(c(1:dim(indels)[1]), function(num) { + ref_seq_test <- as.character(sread(ref_seq)) + if (indels$Modification[[num]] == "del"){ + substr(ref_seq_test, indels$Start[[num]], indels$Start[[num]]+indels$Length[[num]]-1) <- paste0(rep("-", indels$Length[[num]]), collapse = "") + edited_sequences <- c(edited_sequences, ref_seq_test) + } else { + seq <- paste(substring(ref_seq_test, 1, indels$Start[[num]]), paste0(rep("N", indels$Length[[num]]), collapse = ""), substring(ref_seq_test, indels$Start[[num]]+1, nchar(ref_seq_test)), sep = "") + edited_sequences <- c(edited_sequences, seq) + } + }) +} + +###################### +##### Substitutions at -+25 of the gRNA plot +###################### +subs_plot <- function(subsperc, gRNA_seq, cut_site){ + ### Get the gRNA region + pre_cut_site <- cut_site - (nchar(gRNA_seq)-3) ## insted of 17 to allow different gRNA lengths instead of only 20 + post_cut_site <- cut_site + 3 + 1 + ### gRNA nucleotides to use them in the axis + ref_nt <- stringr::str_split(gRNA_seq, "")[[1]] + ### Get the plot + plot <- ggplot(data=subsperc %>% filter(pos > pre_cut_site - 25) %>% filter(pos < post_cut_site + 25), aes(x=ordered(pos), y=percentage, fill=nucleotide, alpha=ifelse(percentage<95,1,0))) + + geom_bar(stat="identity") + theme_classic() + scale_fill_manual("Nuclotides", values = c("A" = "#109648", "C" = "#86b7ed", "G" = "#f7b32b", "T" = "#d62839", "-" = "#f2f2f2")) + + scale_x_discrete(breaks = (pre_cut_site+1):(post_cut_site-1), labels = ref_nt) + xlab(NULL) + scale_alpha(guide = 'none') + ylab('nt (%)') + + geom_text(aes(label = ifelse(percentage<50 & percentage>5,percentage,"")), + hjust = 0, vjust = 1.5, angle = 90, nudge_x = -.5, + size = 2.5) + + theme(text = element_text(size = 11), legend.position = "bottom") + return(plot) +} + +############ +### Substitutions logo plot +############ +subs_logo_plot <- function(subsperc, gRNA_seq, cut_site){ + ### Positions related to the cut site + pre_cut_site <- cut_site - (nchar(gRNA_seq)-3) + post_cut_site <- cut_site + 6 + 1 + # + ### From percentages table to matrix + fper_filtered <- subsperc %>% filter(pos > pre_cut_site) %>% filter(pos < post_cut_site) %>% filter(nucleotide != "-") + nuc_pos <- pivot_wider(fper_filtered, names_from = pos, values_from = percentage) + nuc_pos[is.na(nuc_pos)] <- 0 + nuc_pos[nuc_pos == 100] <- 0 + nuc_pos_matrix <- data.matrix(nuc_pos[,-1]) + nuc_pos_matrix <- nuc_pos_matrix+0.00001 ### This is done to aboid problemes in case of a matrix full of 0s + rownames(nuc_pos_matrix) <- nuc_pos$nucleotide + # + ### gRNA nucleotides to use them in the axis + ref_nt <- c(str_split(gRNA_seq, "")[[1]], "N", "G", "G") + logo <- ggseqlogo(nuc_pos_matrix, method='custom', seq_type='dna') + ylab('nt (%)') + logo$scales$scales[[1]] <- scale_x_continuous(breaks= 1:length(ref_nt),labels=ref_nt) + logo_plot <- logo + annotate('rect', xmin = length(ref_nt)-2.5, xmax = length(ref_nt)+0.5, ymin = -5, ymax = 105, alpha = .1, col='black', fill='yellow') + return(logo_plot) +} + +################## +### Top variants plots +################## + +### Logo plot of top indels +get_logo_top_vars <- function(selected_reference, change_len, pattern_indel, modification, str_pos, cut_site){ + # This function is used to generate the logo of the alleles that apperar most times in data. + all_pre_nt <- list(A = "", C = "", T = "", G = "") + for (j in c("A", "C", "G", "T")){ + per_nt <- lapply(1:length(selected_reference), { + function(i){ + if(i < 11 || i > 10+l || modification == "ins"){ + if(selected_reference[i] == j){ + return(1-0.00001) + } else { + return(0.00001) + } + } else { + return(0.00001) + } + } + }) + all_pre_nt[[j]] <- per_nt + } + ## Data frame with the probability of each position + df<-NULL + df <- rbind(df, unlist(all_pre_nt$A)) + df <- rbind(df, unlist(all_pre_nt$C)) + df <- rbind(df, unlist(all_pre_nt$G)) + df <- rbind(df, unlist(all_pre_nt$T)) + pos_matrix <- data.matrix(df) + rownames(pos_matrix) <- c("A", "C", "G", "T") + if(modification == "ins"){ + cut_correction <- change_len + } else { + cut_correction <- 0 + } + + logo <- ggseqlogo(pos_matrix, method='custom', seq_type='dna') + ylab('') + + theme(plot.title = element_text(size = 12 , face = "bold" , hjust = 0.5), + axis.text.x = element_blank(), + axis.text.y = element_blank()) + + ggtitle(paste0(l, "nt ", mod, "; Start: ", str_pos)) + + annotate('segment', x = cut_site-str_pos+11+0.5+cut_correction , xend=cut_site-str_pos+11+0.5+cut_correction, y=-0.08, yend=1.08, linewidth=1, color="red") + + if (is.na(pattern_indel) && modification != "ins"){ + return(logo) + } else if (modification == "ins"){ + logo_plot <- logo + annotate('rect', xmin = 11-0.5, xmax = 10+change_len+0.5, ymin = -0.05, ymax = 1.05, alpha = .1, col='black', fill='yellow') + return(logo_plot) + } else if (pattern_indel == "NHEJ"){ + return(logo) + } else { + mh_len <- nchar(pattern_indel) + logo_plot <- logo + annotate('rect', xmin = 11+change_len-0.5, xmax = 10+change_len+mh_len+0.5, ymin = -0.05, ymax = 1.05, alpha = .1, col='black', fill='yellow') + return(logo_plot) + } +} + + + +################## +### Get all plots +################## + +#load(subs_info) +#empty data troubleshooter, notice that 3 is choose to avoid counting small empty df product of read.csv function +a<-as.character(data) +sampleName<-gsub('.{11}$', '', a) +checkFaulty<-grep('Faulty', sampleName) +a<-as.character(indels_info) +a<-gsub('.{11}$', '', a) +checkEmpty<-grep('Empt', a) +checkFaulty<-grep('Badl', a) +if (dim(data)[2]>3 && length(checkFaulty) == 0 && length(checkEmpty) == 0){ ### Substitutions percentages plot + cut_site <- get_cutSite(gR, ref_seq, rel_cut_site) + if (dim(subs_plup)[1] == 0){ + system(paste0("touch ", sample_name, "_subs-perc_plot.png")) + } else { + sp <- subs_plot(subs_plup, gR, cut_site) + ggsave(paste0(sample_name, "_subs-perc_plot.png"), width = 12, height = 1.5) + ### Substitutions logo plot + sp_logo <- subs_logo_plot(subs_plup, gR, cut_site) + ggsave(paste0(sample_name, "_subs-perc_plot_LOGO.png"), width = 12, height = 1.5) + } + + ### Counts plot + #GET THE HTMLS + figure_counts <- plotIndels_gg(indels_data = data, cut = cut_site) + htmlwidgets::saveWidget(as_widget(figure_counts[[1]]), paste0(sample_name,"_Deletions.html")) + htmlwidgets::saveWidget(as_widget(figure_counts[[2]]), paste0(sample_name,"_Insertions.html")) + htmlwidgets::saveWidget(as_widget(figure_counts[[3]]), paste0(sample_name,"_accumulative.html")) + + ### Alleles plot (deletions) + indel_count <- indel_count_seq(data) + dels_count <- indel_count %>% filter(Modification == "del") ## Just deletions + if (dim(dels_count)[1] != 0){ + dels_count$Start <- as.numeric(as.character(dels_count$Start)) + dels_count$Length <- as.numeric(as.character(dels_count$Length)) + seqs <- get_sequences(dels_count, ref_seq) + + # Write fasta with the sequences and use it to get the alleles plot + if (length(seqs) < 10){ + rep_seqs <- seqs[1:length(seqs)] + } else { + rep_seqs <- seqs[1:10] + } + write.fasta(sequences = rep_seqs, names = paste0(round(dels_count$Perc, 2), "_", dels_count$Modification, dels_count$Length, "_", seq(1, dim(dels_count)[1]))[1:10], file.out = paste0(sample_name, "_top10.fa")) + nt_sequences <- paste0(sample_name, "_top10.fa") + figure_alleles <- ggmsa(nt_sequences, cut_site-40, cut_site+40, color = "Chemistry_NT", seq_name = TRUE) + ggsave(paste0(sample_name, "_delAlleles_plot.png")) + } else { + system(paste0("touch ", sample_name, "_delAlleles_plot.png")) + } + + ######### Low variability or top variants plot + wt <- data$wt_reads[1] ### 7299 + templata_based <- data$t_reads[1] ### 0 + total_char <- wt + templata_based + dim(data)[1] + + delCols_indels <- data %>% group_by(Modification, Start, Length, ins_nt, patterns) %>% dplyr::summarize(freq = n()) + unique_variants <- rbind(as.data.frame(delCols_indels), c("wt", 0, 0, NA, NA, wt), c("template-based", 0, 0, NA, NA, templata_based)) + uniq_indels_sorted <- unique_variants[order(as.numeric(unique_variants$freq), decreasing = TRUE),] + write.csv(uniq_indels_sorted,file=paste0(sample_name, "_unique-variants.csv")) + + # Check if there are enogh indels to have 5 top + if(dim(uniq_indels_sorted)[1] < 5 ){ + num_top <- dim(delCols_indels)[1] + } else { num_top <- 5 } + + # Get to variants + top_5 <- uniq_indels_sorted[1:num_top,] + + top5_names <- lapply(c(1:dim(top_5)[1]), + function(i){ + if( top_5[i,]$Modification == "del" ) { + return(paste0(top_5[i,]$Start, "_", top_5[i,]$Modification, top_5[i,]$Length, "_", top_5[i,]$pattern)) + } else if ( top_5[i,]$Modification == "ins" ) { + return(paste0(top_5[i,]$Start, "_", top_5[i,]$Modification, top_5[i,]$ins_nt)) + } else { + return(top_5[i,]$Modification) + } + } + ) + + wt_pos <- which(unlist(top5_names) == "wt") + if (length(wt_pos) == 0){ + cols_list = c("#bebebe", rep("#9394f7", num_top + 1)) + } else if (wt_pos == num_top){ + cols_list = c("#bebebe", "#9394f7", rep("#9394f7", num_top - 1), "#1cf453") + } else if (wt_pos == 1) { + cols_list = c("#bebebe", "#9394f7", "#1cf453", rep("#9394f7", num_top - 1)) + } else { + cols_list = c("#bebebe", rep("#9394f7", wt_pos), "#1cf453", rep("#9394f7", num_top-wt_pos)) + } + + reads_classes <- c("Other alleles", "Top alleles", unlist(top5_names)) + reads_counts <- c(total_char - sum(as.numeric(top_5$freq)), sum(as.numeric(top_5$freq)), as.numeric(top_5$freq)) + reads_summary <- data.frame(classes = unlist(reads_classes), counts = unlist(reads_counts)) + reads_summary$parents = c("", "", rep("Top alleles", num_top)) + fig <- plot_ly(reads_summary, + labels = ~ classes, + parents = ~ parents, + values = ~ counts, + type = 'sunburst', + branchvalues = 'total', + textinfo = "label+percent entry", + marker = list(colors = cols_list, color = "black"), + textfont = list(color = '#000000', size = 20) + ) + + htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_top.html")) + + ### Logo plot + all_each_logo <- list() + list_num <- 1 + sel_top <- top_5 %>% filter(Modification %in% c("del", "ins")) + if (dim(sel_top)[1] > 0){ + for (i in c(1:dim(sel_top)[1])){ + if (sel_top[i,]$Modification == "ins"){ + selfil_1 <- data %>% filter(Modification == sel_top[i,]$Modification) %>% filter(Start == sel_top[i,]$Start) %>% filter(Length == sel_top[i,]$Length) %>% filter(ins_nt == sel_top[i,]$ins_nt) + } else { + selfil_1 <- data %>% filter(Modification == sel_top[i,]$Modification) %>% filter(Start == sel_top[i,]$Start) %>% filter(Length == sel_top[i,]$Length) + } + s <- selfil_1[1,]$Start + l <- selfil_1[1,]$Length + if( s > 12 && (nchar(as.character(sread(ref_seq)[[1]])) > (s+l+9) )){ + ref_splited <- c(str_split(toupper(as.character(sread(ref_seq)[[1]])), "")[[1]]) + sel_ref <- ref_splited[(s-10):(s+l+9)] + mod <- selfil_1[1,]$Modification + if (mod == "ins"){ + sel_ref <- c(sel_ref[1:10], c(str_split(toupper(selfil_1[1,]$ins_nt), "")[[1]]), sel_ref[11:length(sel_ref)]) + } + p <- selfil_1[1,]$patterns + each_logo <- get_logo_top_vars(sel_ref, l, p, mod, s, cut_site) + + all_each_logo[[list_num]] <- each_logo + list_num <- list_num + 1 + } + } + } + + if (length(all_each_logo) > 0){ + plot_grid(plotlist = all_each_logo, ncol = 1) + ggsave(paste0(sample_name, "_top-alleles_LOGO.png")) + } else { + system(paste0("touch ", sample_name, "_top-alleles_LOGO.png")) + } + +} else { + fig<-empty_plot("No alignments were produced. + Please check your files and references") + htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_Deletions.html")) + htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_Insertions.html")) + htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_accumulative.html")) + # system(paste0("touch ", sample_name, "_delAlleles_plot.png")) + + #Elements for dynamic table IF THERE ARE NO ALIGNMENTS + empty_list = list() + exportJson <- jsonlite::toJSON(empty_list) + write(exportJson, paste(sample_name,"_length.json",sep="")) + write(exportJson, paste(sample_name,"_Total_reads.json",sep="")) + write(exportJson, paste(sample_name,"_metadels.json",sep="")) + write(exportJson, paste(sample_name,"_metains.json",sep="")) + + system(paste0("touch ", sample_name, "_top.html")) + system(paste0("touch ", sample_name, "_top-alleles_LOGO.png")) + system(paste0("touch ", sample_name, "_counts_plot.png")) + system(paste0("touch ", sample_name, "_subs-perc_plot_LOGO.png")) + system(paste0("touch ", sample_name, "_subs-perc_plot.png")) +} diff --git a/conf/modules.config b/conf/modules.config index f78266e9..8c6b5d6c 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -231,6 +231,10 @@ process { ext.args = '--cut_site=-3' } + withName: CRISPRSEQ_PLOTTER { + ext.args = '--cut_site=-3' + } + withName: CUSTOM_DUMPSOFTWAREVERSIONS { publishDir = [ path: { "${params.outdir}/pipeline_info" }, diff --git a/modules/local/cigar_parser.nf b/modules/local/cigar_parser.nf index 5073ab26..5b87bba4 100644 --- a/modules/local/cigar_parser.nf +++ b/modules/local/cigar_parser.nf @@ -11,7 +11,7 @@ process CIGAR_PARSER { tuple val(meta), path(reads), path(index), path(reference), val(protospacer), path(template), path(template_bam), path(reference_template), path(summary) output: - tuple val(meta), path("*indels.csv"), path("*_subs-perc.csv"), emit: indels + tuple val(meta), path("*[!QC-]indels.csv"), path("*_subs-perc.csv"), emit: indels tuple val(meta), path("*.html"), path("*edits.csv") , emit: edition tuple val(meta), path("*cutSite.json") , emit: cutsite tuple val(meta), path("*_QC-indels.csv") , emit: qcindels diff --git a/modules/local/crisprseq_plotter.nf b/modules/local/crisprseq_plotter.nf new file mode 100644 index 00000000..30a5db68 --- /dev/null +++ b/modules/local/crisprseq_plotter.nf @@ -0,0 +1,42 @@ +process CRISPRSEQ_PLOTTER { + tag "$meta.id" + label 'process_medium' + + conda 'r-ggplot2=3.4.3 bioconductor-shortread=1.58.0 r-ggpubr=0.6.0 r-ggmsa=1.0.2 r-seqmagick=0.1.6 r-tidyr=1.3.0 r-ggseqlogo=0.1 r-cowplot=1.1.1 r-seqinr=4.2_30 r-optparse=1.7.3 r-dplyr=1.1.2 r-plyr=1.8.8 r-stringr=1.5.0 r-plotly=4.10.2' + // TODO: change containers + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/mulled-v2-61c59287265e27f2c4589cfc90013ef6c2c6acf1:fb3e48060a8c0e5108b1b60a2ad7e090cfb9eee5-0' : + 'biocontainers/mulled-v2-61c59287265e27f2c4589cfc90013ef6c2c6acf1:fb3e48060a8c0e5108b1b60a2ad7e090cfb9eee5-0' }" + + input: + tuple val(meta), path(indels), path(substitutions), path(reference), val(protospacer) + + output: + tuple val(meta), path("*_Deletions.html"), path("*_delAlleles_plot.png") , emit: deletions + tuple val(meta), path("*_Insertions.html") , emit: insertions + tuple val(meta), path("*_accumulative.html") , emit: accumulative + tuple val(meta), path("*_subs-perc_plot.png"), path("*_subs-perc_plot_LOGO.png"), emit: substitutions + tuple val(meta), path("*_top-alleles_LOGO.png"), path("*_top.html") , emit: topalleles + 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}" + """ + plotter.R \\ + $args \\ + --indels_info=$indels \\ + --reference=$reference \\ + --gRNA_sequence=$protospacer \\ + --sample_name=$prefix \\ + --substitutions_info=$substitutions + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + Rscript: \$(Rscript --version) + END_VERSIONS + """ +} diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 15d870a0..33c2ee59 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -46,6 +46,7 @@ include { PREPROCESSING_SUMMARY } from '../modules/local/preprocessing_summary' include { CLUSTERING_SUMMARY } from '../modules/local/clustering_summary' include { ALIGNMENT_SUMMARY } from '../modules/local/alignment_summary' include { TEMPLATE_REFERENCE } from '../modules/local/template_reference' +include { CRISPRSEQ_PLOTTER } from '../modules/local/crisprseq_plotter' /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -706,12 +707,7 @@ workflow CRISPRSEQ_TARGETED { ch_mapped_bam .join(SAMTOOLS_INDEX.out.bai) .join(ORIENT_REFERENCE.out.reference) - .join(ch_input.protospacer - .map { - meta, fastq -> - [ meta , fastq ] - } - ) + .join(ch_input.protospacer) .join(ch_seq_template, remainder: true) .join(ch_template_bam, remainder: true) .join(TEMPLATE_REFERENCE.out.fasta, remainder: true) @@ -740,6 +736,17 @@ workflow CRISPRSEQ_TARGETED { ch_versions = ch_versions.mix(CIGAR_PARSER.out.versions.first()) + // + // + // + CRISPRSEQ_PLOTTER ( + CIGAR_PARSER.out.indels + .join(ORIENT_REFERENCE.out.reference) + .join(ch_input.protospacer) + ) + ch_versions = ch_versions.mix(CRISPRSEQ_PLOTTER.out.versions.first()) + + // // MODULE: Dump software versions // From c546e5403aae0891ac61428a0ee2e8a35fe76fc4 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Fri, 18 Aug 2023 17:29:46 +0200 Subject: [PATCH 063/133] save plot results in a folder called plots --- conf/modules.config | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/conf/modules.config b/conf/modules.config index 8c6b5d6c..b282aebc 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -233,6 +233,11 @@ process { withName: CRISPRSEQ_PLOTTER { ext.args = '--cut_site=-3' + publishDir = [ + path: { "${params.outdir}/plots" }, + mode: params.publish_dir_mode, + saveAs: { filename -> filename.equals('versions.yml') ? null : filename } + ] } withName: CUSTOM_DUMPSOFTWAREVERSIONS { From 7ea4c31759af1c2deb1caa21c0e00a0e6b6bdce2 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 21 Aug 2023 14:46:17 +0200 Subject: [PATCH 064/133] print plotted percentage in substitution percentage plot --- bin/plotter.R | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/bin/plotter.R b/bin/plotter.R index 10ab023d..883b2105 100755 --- a/bin/plotter.R +++ b/bin/plotter.R @@ -400,7 +400,7 @@ subs_plot <- function(subsperc, gRNA_seq, cut_site){ plot <- ggplot(data=subsperc %>% filter(pos > pre_cut_site - 25) %>% filter(pos < post_cut_site + 25), aes(x=ordered(pos), y=percentage, fill=nucleotide, alpha=ifelse(percentage<95,1,0))) + geom_bar(stat="identity") + theme_classic() + scale_fill_manual("Nuclotides", values = c("A" = "#109648", "C" = "#86b7ed", "G" = "#f7b32b", "T" = "#d62839", "-" = "#f2f2f2")) + scale_x_discrete(breaks = (pre_cut_site+1):(post_cut_site-1), labels = ref_nt) + xlab(NULL) + scale_alpha(guide = 'none') + ylab('nt (%)') + - geom_text(aes(label = ifelse(percentage<50 & percentage>5,percentage,"")), + geom_text(aes(label = ifelse(percentage<50 & percentage>5,100-percentage,"")), hjust = 0, vjust = 1.5, angle = 90, nudge_x = -.5, size = 2.5) + theme(text = element_text(size = 11), legend.position = "bottom") @@ -627,7 +627,7 @@ if (dim(data)[2]>3 && length(checkFaulty) == 0 && length(checkEmpty) == 0){ ### } p <- selfil_1[1,]$patterns each_logo <- get_logo_top_vars(sel_ref, l, p, mod, s, cut_site) - + all_each_logo[[list_num]] <- each_logo list_num <- list_num + 1 } From 6f57715525385dede52608347976314a99af0fb0 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 21 Aug 2023 14:46:54 +0200 Subject: [PATCH 065/133] add documentation of output plots --- docs/output/targeted.md | 26 ++++++++++++++++++++++++++ 1 file changed, 26 insertions(+) diff --git a/docs/output/targeted.md b/docs/output/targeted.md index 0a4ba78e..f44ffd0b 100644 --- a/docs/output/targeted.md +++ b/docs/output/targeted.md @@ -33,6 +33,7 @@ The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes d - [bowtie2](#bowtie2) - Mapping reads to reference - [Edits calling](#edits-calling) - [CIGAR](#cigar) - Parse CIGAR to call edits + - [Output plots](#output-plots) - Results visualisation - [MultiQC](#multiqc) - Aggregate report describing results and QC from the whole pipeline - [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution @@ -262,6 +263,31 @@ This section contains the final output of the pipeline. It contains information
      +### Output plots + +
      +Output files + +- `plots/` + - `*_accumulative.html`: Interactive barplot showing the accumulative deletions and insertions. x-axis represents the reference position. y-axis represents the percentage of reads containing a deletion or insertion in that position. + ![Test sample hCas9-AAVS1-a accumulative edition plot](../images/hCas9-AAVS1-a_accumulative.html) + - `*_delAlleles_plot.png`: Image showing the most common deletions found. x-axis represents the position. y-axis indicates the percentage in which the plotted deletion is observed (in respect of all deletions), followed by the length of the deletion. Dashes `-` indicate a deleted base. + ![Test sample hCas9-AAVS1-a deletion alleles plot](../images/hCas9-AAVS1-a_delAlleles_plot.png) + - `*_Deletions.html`: Interactive barplot showing the percentage of reads showing a deltion for each position and the deletion sizes. The left panel represents the percentage of reads having a deletion for each position (similar to `*_accumulative.html`). The right panel shows the number of deletions found relative to their size. The deleted sequences found are shown coloured in the stacked barplot. + ![Test sample hCas9-AAVS1-a deletions plot](../images/hCas9-AAVS1-a_Deletions.html) + - `*_Insertions.html`: Interactive barplot showing the percentage of reads showing an insertion for each position and the intertion sizes. The left panel represents the percentage of reads having an insertion for each position (similar to `*_accumulative.html`). The right panel shows the number of insertions found relative to their size. The inserted sequences found are shown coloured in the stacked barplot. + ![Test sample hCas9-AAVS1-a insertions plot](../images/hCas9-AAVS1-a_Insertions.html) + - `*_subs-perc_plot_LOGO.png`: LOGO showing the most represented nucleotide and its percentage (y-axis) for protospacer positions. PAM sequence is highlighted in yellow. + ![Test sample hCas9-AAVS1-a substitutions LOGO](../images/hCas9-AAVS1-a_subs-perc_plot_LOGO.png) + - `*_subs-perc_plot.png`: Barplot showing the most represented nucleotide and it's percentage (y-axis and bar tags) for +/-25 positions surrounding the cut site. The protospacer sequence is highlighted by writing the sequence base in the y axis. Bases whose percentage is higher than 90% are not colored. + ![Test sample hCas9-AAVS1-a substitutions percentage plot](../images/hCas9-AAVS1-a_subs-perc_plot.png) + - `*_top-alleles_LOGO.png`: LOGO showing the 4 most represented editions. Cut site is highlighted with a vertical red line. The type of edition and start position are shown as a title to each LOGO. Deleted bases are not drawn. Inserted bases are highlighted in yellow. + ![Test sample hCas9-AAVS1-a top alleles LOGO](../images/hCas9-AAVS1-a_top-alleles_LOGO.png) + - `*_top.html`: Interactive pie chart showing the percentage of the top 4 editions found. The percentage of WT is also shown. Editions are named after the position, the type of edition and length and the sequence. + ![Test sample hCas9-AAVS1-a top alleles plot](../images/hCas9-AAVS1-a_top.html) + +
      + ## MultiQC
      From 11cfda2a2318f9356b5087797c8c16256c802411 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 21 Aug 2023 15:01:44 +0200 Subject: [PATCH 066/133] add images of output example plots --- docs/images/hCas9-AAVS1-a_Deletions.png | Bin 0 -> 183460 bytes docs/images/hCas9-AAVS1-a_Insertions.png | Bin 0 -> 88166 bytes docs/images/hCas9-AAVS1-a_accumulative.png | Bin 0 -> 139340 bytes docs/images/hCas9-AAVS1-a_delAlleles_plot.png | Bin 0 -> 372550 bytes docs/images/hCas9-AAVS1-a_subs-perc_plot.png | Bin 0 -> 174001 bytes .../hCas9-AAVS1-a_subs-perc_plot_LOGO.png | Bin 0 -> 155293 bytes docs/images/hCas9-AAVS1-a_top-alleles_LOGO.png | Bin 0 -> 475593 bytes docs/images/hCas9-AAVS1-a_top.png | Bin 0 -> 130763 bytes docs/output/targeted.md | 8 ++++---- 9 files changed, 4 insertions(+), 4 deletions(-) create mode 100644 docs/images/hCas9-AAVS1-a_Deletions.png create mode 100644 docs/images/hCas9-AAVS1-a_Insertions.png create mode 100644 docs/images/hCas9-AAVS1-a_accumulative.png create 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It contains information - `plots/` - `*_accumulative.html`: Interactive barplot showing the accumulative deletions and insertions. x-axis represents the reference position. y-axis represents the percentage of reads containing a deletion or insertion in that position. - ![Test sample hCas9-AAVS1-a accumulative edition plot](../images/hCas9-AAVS1-a_accumulative.html) + ![Test sample hCas9-AAVS1-a accumulative edition plot](../images/hCas9-AAVS1-a_accumulative.png) - `*_delAlleles_plot.png`: Image showing the most common deletions found. x-axis represents the position. y-axis indicates the percentage in which the plotted deletion is observed (in respect of all deletions), followed by the length of the deletion. Dashes `-` indicate a deleted base. ![Test sample hCas9-AAVS1-a deletion alleles plot](../images/hCas9-AAVS1-a_delAlleles_plot.png) - `*_Deletions.html`: Interactive barplot showing the percentage of reads showing a deltion for each position and the deletion sizes. The left panel represents the percentage of reads having a deletion for each position (similar to `*_accumulative.html`). The right panel shows the number of deletions found relative to their size. The deleted sequences found are shown coloured in the stacked barplot. - ![Test sample hCas9-AAVS1-a deletions plot](../images/hCas9-AAVS1-a_Deletions.html) + ![Test sample hCas9-AAVS1-a deletions plot](../images/hCas9-AAVS1-a_Deletions.png) - `*_Insertions.html`: Interactive barplot showing the percentage of reads showing an insertion for each position and the intertion sizes. The left panel represents the percentage of reads having an insertion for each position (similar to `*_accumulative.html`). The right panel shows the number of insertions found relative to their size. The inserted sequences found are shown coloured in the stacked barplot. - ![Test sample hCas9-AAVS1-a insertions plot](../images/hCas9-AAVS1-a_Insertions.html) + ![Test sample hCas9-AAVS1-a insertions plot](../images/hCas9-AAVS1-a_Insertions.png) - `*_subs-perc_plot_LOGO.png`: LOGO showing the most represented nucleotide and its percentage (y-axis) for protospacer positions. PAM sequence is highlighted in yellow. ![Test sample hCas9-AAVS1-a substitutions LOGO](../images/hCas9-AAVS1-a_subs-perc_plot_LOGO.png) - `*_subs-perc_plot.png`: Barplot showing the most represented nucleotide and it's percentage (y-axis and bar tags) for +/-25 positions surrounding the cut site. The protospacer sequence is highlighted by writing the sequence base in the y axis. Bases whose percentage is higher than 90% are not colored. @@ -284,7 +284,7 @@ This section contains the final output of the pipeline. It contains information - `*_top-alleles_LOGO.png`: LOGO showing the 4 most represented editions. Cut site is highlighted with a vertical red line. The type of edition and start position are shown as a title to each LOGO. Deleted bases are not drawn. Inserted bases are highlighted in yellow. ![Test sample hCas9-AAVS1-a top alleles LOGO](../images/hCas9-AAVS1-a_top-alleles_LOGO.png) - `*_top.html`: Interactive pie chart showing the percentage of the top 4 editions found. The percentage of WT is also shown. Editions are named after the position, the type of edition and length and the sequence. - ![Test sample hCas9-AAVS1-a top alleles plot](../images/hCas9-AAVS1-a_top.html) + ![Test sample hCas9-AAVS1-a top alleles plot](../images/hCas9-AAVS1-a_top.png)

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      From 4acfc21892511cda60db6a8a5c358dc10f317465 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 22 Aug 2023 08:52:30 +0200 Subject: [PATCH 067/133] add mulled container for crisprseq_plotter --- modules/local/crisprseq_plotter.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/crisprseq_plotter.nf b/modules/local/crisprseq_plotter.nf index 30a5db68..2a368f6c 100644 --- a/modules/local/crisprseq_plotter.nf +++ b/modules/local/crisprseq_plotter.nf @@ -5,8 +5,8 @@ process CRISPRSEQ_PLOTTER { conda 'r-ggplot2=3.4.3 bioconductor-shortread=1.58.0 r-ggpubr=0.6.0 r-ggmsa=1.0.2 r-seqmagick=0.1.6 r-tidyr=1.3.0 r-ggseqlogo=0.1 r-cowplot=1.1.1 r-seqinr=4.2_30 r-optparse=1.7.3 r-dplyr=1.1.2 r-plyr=1.8.8 r-stringr=1.5.0 r-plotly=4.10.2' // TODO: change containers container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/mulled-v2-61c59287265e27f2c4589cfc90013ef6c2c6acf1:fb3e48060a8c0e5108b1b60a2ad7e090cfb9eee5-0' : - 'biocontainers/mulled-v2-61c59287265e27f2c4589cfc90013ef6c2c6acf1:fb3e48060a8c0e5108b1b60a2ad7e090cfb9eee5-0' }" + 'https://depot.galaxyproject.org/singularity/mulled-v2-6de07928379e6eface08a0019c4a1d6b5192e805:0d77388f37ddd923a087f7792e30e83ab54c918c-0' : + 'biocontainers/mulled-v2-6de07928379e6eface08a0019c4a1d6b5192e805:0d77388f37ddd923a087f7792e30e83ab54c918c-0' }" input: tuple val(meta), path(indels), path(substitutions), path(reference), val(protospacer) From a6cfae3ebde3b9f1d3e771664683da76a6bf8b67 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 22 Aug 2023 09:19:37 +0200 Subject: [PATCH 068/133] fix left-padding spaces --- bin/plotter.R | 220 +++++++++++++++++++++++++------------------------- 1 file changed, 110 insertions(+), 110 deletions(-) diff --git a/bin/plotter.R b/bin/plotter.R index 883b2105..33a008f2 100755 --- a/bin/plotter.R +++ b/bin/plotter.R @@ -547,119 +547,119 @@ if (dim(data)[2]>3 && length(checkFaulty) == 0 && length(checkEmpty) == 0){ ### system(paste0("touch ", sample_name, "_delAlleles_plot.png")) } - ######### Low variability or top variants plot - wt <- data$wt_reads[1] ### 7299 - templata_based <- data$t_reads[1] ### 0 - total_char <- wt + templata_based + dim(data)[1] - - delCols_indels <- data %>% group_by(Modification, Start, Length, ins_nt, patterns) %>% dplyr::summarize(freq = n()) - unique_variants <- rbind(as.data.frame(delCols_indels), c("wt", 0, 0, NA, NA, wt), c("template-based", 0, 0, NA, NA, templata_based)) - uniq_indels_sorted <- unique_variants[order(as.numeric(unique_variants$freq), decreasing = TRUE),] - write.csv(uniq_indels_sorted,file=paste0(sample_name, "_unique-variants.csv")) - - # Check if there are enogh indels to have 5 top - if(dim(uniq_indels_sorted)[1] < 5 ){ - num_top <- dim(delCols_indels)[1] - } else { num_top <- 5 } - - # Get to variants - top_5 <- uniq_indels_sorted[1:num_top,] - - top5_names <- lapply(c(1:dim(top_5)[1]), - function(i){ - if( top_5[i,]$Modification == "del" ) { - return(paste0(top_5[i,]$Start, "_", top_5[i,]$Modification, top_5[i,]$Length, "_", top_5[i,]$pattern)) - } else if ( top_5[i,]$Modification == "ins" ) { - return(paste0(top_5[i,]$Start, "_", top_5[i,]$Modification, top_5[i,]$ins_nt)) - } else { - return(top_5[i,]$Modification) - } - } - ) - - wt_pos <- which(unlist(top5_names) == "wt") - if (length(wt_pos) == 0){ - cols_list = c("#bebebe", rep("#9394f7", num_top + 1)) - } else if (wt_pos == num_top){ - cols_list = c("#bebebe", "#9394f7", rep("#9394f7", num_top - 1), "#1cf453") - } else if (wt_pos == 1) { - cols_list = c("#bebebe", "#9394f7", "#1cf453", rep("#9394f7", num_top - 1)) - } else { - cols_list = c("#bebebe", rep("#9394f7", wt_pos), "#1cf453", rep("#9394f7", num_top-wt_pos)) - } - - reads_classes <- c("Other alleles", "Top alleles", unlist(top5_names)) - reads_counts <- c(total_char - sum(as.numeric(top_5$freq)), sum(as.numeric(top_5$freq)), as.numeric(top_5$freq)) - reads_summary <- data.frame(classes = unlist(reads_classes), counts = unlist(reads_counts)) - reads_summary$parents = c("", "", rep("Top alleles", num_top)) - fig <- plot_ly(reads_summary, - labels = ~ classes, - parents = ~ parents, - values = ~ counts, - type = 'sunburst', - branchvalues = 'total', - textinfo = "label+percent entry", - marker = list(colors = cols_list, color = "black"), - textfont = list(color = '#000000', size = 20) - ) - - htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_top.html")) - - ### Logo plot - all_each_logo <- list() - list_num <- 1 - sel_top <- top_5 %>% filter(Modification %in% c("del", "ins")) - if (dim(sel_top)[1] > 0){ - for (i in c(1:dim(sel_top)[1])){ - if (sel_top[i,]$Modification == "ins"){ - selfil_1 <- data %>% filter(Modification == sel_top[i,]$Modification) %>% filter(Start == sel_top[i,]$Start) %>% filter(Length == sel_top[i,]$Length) %>% filter(ins_nt == sel_top[i,]$ins_nt) - } else { - selfil_1 <- data %>% filter(Modification == sel_top[i,]$Modification) %>% filter(Start == sel_top[i,]$Start) %>% filter(Length == sel_top[i,]$Length) - } - s <- selfil_1[1,]$Start - l <- selfil_1[1,]$Length - if( s > 12 && (nchar(as.character(sread(ref_seq)[[1]])) > (s+l+9) )){ - ref_splited <- c(str_split(toupper(as.character(sread(ref_seq)[[1]])), "")[[1]]) - sel_ref <- ref_splited[(s-10):(s+l+9)] - mod <- selfil_1[1,]$Modification - if (mod == "ins"){ - sel_ref <- c(sel_ref[1:10], c(str_split(toupper(selfil_1[1,]$ins_nt), "")[[1]]), sel_ref[11:length(sel_ref)]) - } - p <- selfil_1[1,]$patterns - each_logo <- get_logo_top_vars(sel_ref, l, p, mod, s, cut_site) + ######### Low variability or top variants plot + wt <- data$wt_reads[1] ### 7299 + templata_based <- data$t_reads[1] ### 0 + total_char <- wt + templata_based + dim(data)[1] + + delCols_indels <- data %>% group_by(Modification, Start, Length, ins_nt, patterns) %>% dplyr::summarize(freq = n()) + unique_variants <- rbind(as.data.frame(delCols_indels), c("wt", 0, 0, NA, NA, wt), c("template-based", 0, 0, NA, NA, templata_based)) + uniq_indels_sorted <- unique_variants[order(as.numeric(unique_variants$freq), decreasing = TRUE),] + write.csv(uniq_indels_sorted,file=paste0(sample_name, "_unique-variants.csv")) + + # Check if there are enogh indels to have 5 top + if(dim(uniq_indels_sorted)[1] < 5 ){ + num_top <- dim(delCols_indels)[1] + } else { num_top <- 5 } + + # Get to variants + top_5 <- uniq_indels_sorted[1:num_top,] + + top5_names <- lapply(c(1:dim(top_5)[1]), + function(i){ + if( top_5[i,]$Modification == "del" ) { + return(paste0(top_5[i,]$Start, "_", top_5[i,]$Modification, top_5[i,]$Length, "_", top_5[i,]$pattern)) + } else if ( top_5[i,]$Modification == "ins" ) { + return(paste0(top_5[i,]$Start, "_", top_5[i,]$Modification, top_5[i,]$ins_nt)) + } else { + return(top_5[i,]$Modification) + } + } + ) + + wt_pos <- which(unlist(top5_names) == "wt") + if (length(wt_pos) == 0){ + cols_list = c("#bebebe", rep("#9394f7", num_top + 1)) + } else if (wt_pos == num_top){ + cols_list = c("#bebebe", "#9394f7", rep("#9394f7", num_top - 1), "#1cf453") + } else if (wt_pos == 1) { + cols_list = c("#bebebe", "#9394f7", "#1cf453", rep("#9394f7", num_top - 1)) + } else { + cols_list = c("#bebebe", rep("#9394f7", wt_pos), "#1cf453", rep("#9394f7", num_top-wt_pos)) + } - all_each_logo[[list_num]] <- each_logo - list_num <- list_num + 1 - } + reads_classes <- c("Other alleles", "Top alleles", unlist(top5_names)) + reads_counts <- c(total_char - sum(as.numeric(top_5$freq)), sum(as.numeric(top_5$freq)), as.numeric(top_5$freq)) + reads_summary <- data.frame(classes = unlist(reads_classes), counts = unlist(reads_counts)) + reads_summary$parents = c("", "", rep("Top alleles", num_top)) + fig <- plot_ly(reads_summary, + labels = ~ classes, + parents = ~ parents, + values = ~ counts, + type = 'sunburst', + branchvalues = 'total', + textinfo = "label+percent entry", + marker = list(colors = cols_list, color = "black"), + textfont = list(color = '#000000', size = 20) + ) + + htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_top.html")) + + ### Logo plot + all_each_logo <- list() + list_num <- 1 + sel_top <- top_5 %>% filter(Modification %in% c("del", "ins")) + if (dim(sel_top)[1] > 0){ + for (i in c(1:dim(sel_top)[1])){ + if (sel_top[i,]$Modification == "ins"){ + selfil_1 <- data %>% filter(Modification == sel_top[i,]$Modification) %>% filter(Start == sel_top[i,]$Start) %>% filter(Length == sel_top[i,]$Length) %>% filter(ins_nt == sel_top[i,]$ins_nt) + } else { + selfil_1 <- data %>% filter(Modification == sel_top[i,]$Modification) %>% filter(Start == sel_top[i,]$Start) %>% filter(Length == sel_top[i,]$Length) + } + s <- selfil_1[1,]$Start + l <- selfil_1[1,]$Length + if( s > 12 && (nchar(as.character(sread(ref_seq)[[1]])) > (s+l+9) )){ + ref_splited <- c(str_split(toupper(as.character(sread(ref_seq)[[1]])), "")[[1]]) + sel_ref <- ref_splited[(s-10):(s+l+9)] + mod <- selfil_1[1,]$Modification + if (mod == "ins"){ + sel_ref <- c(sel_ref[1:10], c(str_split(toupper(selfil_1[1,]$ins_nt), "")[[1]]), sel_ref[11:length(sel_ref)]) + } + p <- selfil_1[1,]$patterns + each_logo <- get_logo_top_vars(sel_ref, l, p, mod, s, cut_site) + + all_each_logo[[list_num]] <- each_logo + list_num <- list_num + 1 + } + } } - } - if (length(all_each_logo) > 0){ - plot_grid(plotlist = all_each_logo, ncol = 1) - ggsave(paste0(sample_name, "_top-alleles_LOGO.png")) - } else { - system(paste0("touch ", sample_name, "_top-alleles_LOGO.png")) - } + if (length(all_each_logo) > 0){ + plot_grid(plotlist = all_each_logo, ncol = 1) + ggsave(paste0(sample_name, "_top-alleles_LOGO.png")) + } else { + system(paste0("touch ", sample_name, "_top-alleles_LOGO.png")) + } } else { - fig<-empty_plot("No alignments were produced. - Please check your files and references") - htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_Deletions.html")) - htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_Insertions.html")) - htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_accumulative.html")) - # system(paste0("touch ", sample_name, "_delAlleles_plot.png")) - - #Elements for dynamic table IF THERE ARE NO ALIGNMENTS - empty_list = list() - exportJson <- jsonlite::toJSON(empty_list) - write(exportJson, paste(sample_name,"_length.json",sep="")) - write(exportJson, paste(sample_name,"_Total_reads.json",sep="")) - write(exportJson, paste(sample_name,"_metadels.json",sep="")) - write(exportJson, paste(sample_name,"_metains.json",sep="")) - - system(paste0("touch ", sample_name, "_top.html")) - system(paste0("touch ", sample_name, "_top-alleles_LOGO.png")) - system(paste0("touch ", sample_name, "_counts_plot.png")) - system(paste0("touch ", sample_name, "_subs-perc_plot_LOGO.png")) - system(paste0("touch ", sample_name, "_subs-perc_plot.png")) + fig<-empty_plot("No alignments were produced. + Please check your files and references") + htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_Deletions.html")) + htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_Insertions.html")) + htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_accumulative.html")) + # system(paste0("touch ", sample_name, "_delAlleles_plot.png")) + + #Elements for dynamic table IF THERE ARE NO ALIGNMENTS + empty_list = list() + exportJson <- jsonlite::toJSON(empty_list) + write(exportJson, paste(sample_name,"_length.json",sep="")) + write(exportJson, paste(sample_name,"_Total_reads.json",sep="")) + write(exportJson, paste(sample_name,"_metadels.json",sep="")) + write(exportJson, paste(sample_name,"_metains.json",sep="")) + + system(paste0("touch ", sample_name, "_top.html")) + system(paste0("touch ", sample_name, "_top-alleles_LOGO.png")) + system(paste0("touch ", sample_name, "_counts_plot.png")) + system(paste0("touch ", sample_name, "_subs-perc_plot_LOGO.png")) + system(paste0("touch ", sample_name, "_subs-perc_plot.png")) } From 25141457a6e1c29e8a1ffe98ae1f7ac3045e8a18 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 22 Aug 2023 10:30:59 +0200 Subject: [PATCH 069/133] output empty *_delAlleles_plot.png image when required --- bin/plotter.R | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/bin/plotter.R b/bin/plotter.R index 33a008f2..0982f5e8 100755 --- a/bin/plotter.R +++ b/bin/plotter.R @@ -647,7 +647,7 @@ if (dim(data)[2]>3 && length(checkFaulty) == 0 && length(checkEmpty) == 0){ ### htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_Deletions.html")) htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_Insertions.html")) htmlwidgets::saveWidget(as_widget(fig), paste0(sample_name,"_accumulative.html")) - # system(paste0("touch ", sample_name, "_delAlleles_plot.png")) + system(paste0("touch ", sample_name, "_delAlleles_plot.png")) #Elements for dynamic table IF THERE ARE NO ALIGNMENTS empty_list = list() From fafdc8265129317c2d58ba7b78c0bbf53772d5e5 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 22 Aug 2023 10:55:11 +0200 Subject: [PATCH 070/133] fix modules bwa_index and bowtie2_build after module update --- CHANGELOG.md | 1 + workflows/crisprseq_targeted.nf | 4 ++-- 2 files changed, 3 insertions(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index e31b08b8..48fc69c6 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -16,6 +16,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - Summary processes don't modify the input file anymore, allowing resuming these processes ([#66](https://github.com/nf-core/crisprseq/pull/66)) - Do not stash unexistent files, use empty lists instead. Fixes AWS tests ([#67](https://github.com/nf-core/crisprseq/pull/67)) - Rename process `merging_summary` to `preprocessing_summary` to improve clarity ([#69](https://github.com/nf-core/crisprseq/pull/69)) +- Fix modules `BWA_INDEX` and `BOWTIE2_BUILD` after module update, new versions accept a meta map ### Deprecated diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 15d870a0..8792469e 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -621,7 +621,7 @@ workflow CRISPRSEQ_TARGETED { // if (params.aligner == "bwa") { BWA_INDEX ( - ORIENT_REFERENCE.out.reference.map { it[1] } + ORIENT_REFERENCE.out.reference ) ch_versions = ch_versions.mix(BWA_INDEX.out.versions) BWA_MEM ( @@ -638,7 +638,7 @@ workflow CRISPRSEQ_TARGETED { // if (params.aligner == "bowtie2") { BOWTIE2_BUILD ( - ORIENT_REFERENCE.out.reference.map { it[1] } + ORIENT_REFERENCE.out.reference ) ch_versions = ch_versions.mix(BOWTIE2_BUILD.out.versions) BOWTIE2_ALIGN ( From a5553e59147d9c25e2cf68b9e921380c06cc7a32 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 22 Aug 2023 10:59:33 +0200 Subject: [PATCH 071/133] update changelog --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 48fc69c6..9585c977 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -16,7 +16,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - Summary processes don't modify the input file anymore, allowing resuming these processes ([#66](https://github.com/nf-core/crisprseq/pull/66)) - Do not stash unexistent files, use empty lists instead. Fixes AWS tests ([#67](https://github.com/nf-core/crisprseq/pull/67)) - Rename process `merging_summary` to `preprocessing_summary` to improve clarity ([#69](https://github.com/nf-core/crisprseq/pull/69)) -- Fix modules `BWA_INDEX` and `BOWTIE2_BUILD` after module update, new versions accept a meta map +- Fix modules `BWA_INDEX` and `BOWTIE2_BUILD` after module update, new versions accept a meta map ([#76](https://github.com/nf-core/crisprseq/pull/76)) ### Deprecated From 08334c0aaaa895494a4317c38afe6a1c016a7ec9 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 22 Aug 2023 11:23:10 +0200 Subject: [PATCH 072/133] output empty *_subs-perc_plot_LOGO.png image when required --- bin/plotter.R | 1 + 1 file changed, 1 insertion(+) diff --git a/bin/plotter.R b/bin/plotter.R index 0982f5e8..fb540e7f 100755 --- a/bin/plotter.R +++ b/bin/plotter.R @@ -510,6 +510,7 @@ if (dim(data)[2]>3 && length(checkFaulty) == 0 && length(checkEmpty) == 0){ ### cut_site <- get_cutSite(gR, ref_seq, rel_cut_site) if (dim(subs_plup)[1] == 0){ system(paste0("touch ", sample_name, "_subs-perc_plot.png")) + system(paste0("touch ", sample_name, "_subs-perc_plot_LOGO.png")) } else { sp <- subs_plot(subs_plup, gR, cut_site) ggsave(paste0(sample_name, "_subs-perc_plot.png"), width = 12, height = 1.5) From 0d7aeb44288db053a787cd0fd9e7f33023372be3 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 23 Aug 2023 09:02:34 +0200 Subject: [PATCH 073/133] Update workflows/crisprseq_screening.nf MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- workflows/crisprseq_screening.nf | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 54a2d3b0..a037ad13 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -15,8 +15,7 @@ log.info logo + paramsSummaryLog(workflow) + citation WorkflowCrisprseq.initialise(params, log) -// Check mandatory parameters -//if (!params.count_table) { ch_input = file(params.input) } else { error('Input samplesheet not specified!') } +// Set screening parameters and channels if (params.library) { ch_library = file(params.library) } if (params.crisprcleanr) { ch_crisprcleanr = Channel.value(params.crisprcleanr) } From 6b75dd93eda477e3bee01a6d72e3439bce64acdf Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Thu, 24 Aug 2023 13:43:37 +0200 Subject: [PATCH 074/133] Add license --- modules/local/bagel2/graph.nf | 38 +++++++++++++++++++++++++++++++++++ 1 file changed, 38 insertions(+) diff --git a/modules/local/bagel2/graph.nf b/modules/local/bagel2/graph.nf index 8ca0d84c..26b404ab 100644 --- a/modules/local/bagel2/graph.nf +++ b/modules/local/bagel2/graph.nf @@ -22,9 +22,47 @@ process BAGEL2_GRAPH { def args = task.ext.args ?: '' def prefix = task.ext.prefix ?: "${meta.id}" + /* + # --------------------------------- + # BAGEL: Bayesian Analysis of Gene EssentaLity + # (c) Traver Hart , Eiru Kim 2017. + + # Acknowledgements: John McGonigle + # modified 10/2019 + # Free to modify and redistribute with attribution + # --------------------------------- + + # ------------------------------------ + # constants + +""" + MIT License + + Copyright (c) 2020 Hart Lab + + Permission is hereby granted, free of charge, to any person obtaining a copy + of this software and associated documentation files (the "Software"), to deal + in the Software without restriction, including without limitation the rights + to use, copy, modify, merge, publish, distribute, sublicense, and/or sell + copies of the Software, and to permit persons to whom the Software is + furnished to do so, subject to the following conditions: + + The above copyright notice and this permission notice shall be included in all + copies or substantial portions of the Software. + + THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR + IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, + FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE + AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER + LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, + OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE + SOFTWARE. + + */ """ #!/usr/bin/env python3 + import pandas as pd import matplotlib.pyplot as plt From 0ca0b702bcb35b0e97ccbb7e307548ad6b703141 Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Mon, 25 Sep 2023 15:16:13 +0000 Subject: [PATCH 075/133] Template update for nf-core/tools version 2.10 --- .devcontainer/devcontainer.json | 1 + .github/CONTRIBUTING.md | 4 +- .github/workflows/linting.yml | 2 +- .github/workflows/release-announcments.yml | 68 +++++++++ CHANGELOG.md | 2 +- CITATIONS.md | 2 +- CODE_OF_CONDUCT.md | 133 ++++++++++++++---- LICENSE | 2 +- README.md | 23 +-- assets/email_template.html | 2 +- assets/multiqc_config.yml | 4 +- conf/modules.config | 9 ++ docs/output.md | 5 +- docs/usage.md | 16 ++- lib/NfcoreTemplate.groovy | 16 +++ lib/WorkflowCrisprseq.groovy | 2 +- main.nf | 3 + modules.json | 6 +- .../custom/dumpsoftwareversions/main.nf | 2 +- modules/nf-core/fastqc/main.nf | 8 +- modules/nf-core/multiqc/main.nf | 2 +- nextflow.config | 13 +- nextflow_schema.json | 17 +-- workflows/crisprseq.nf | 1 + 24 files changed, 259 insertions(+), 84 deletions(-) create mode 100644 .github/workflows/release-announcments.yml diff --git a/.devcontainer/devcontainer.json b/.devcontainer/devcontainer.json index ea27a584..4ecfbfe3 100644 --- a/.devcontainer/devcontainer.json +++ b/.devcontainer/devcontainer.json @@ -2,6 +2,7 @@ "name": "nfcore", "image": "nfcore/gitpod:latest", "remoteUser": "gitpod", + "runArgs": ["--privileged"], // Configure tool-specific properties. "customizations": { diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index 607441a1..212cbdbe 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -9,7 +9,9 @@ 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 ;) -> If you need help using or modifying nf-core/crisprseq then the best place to ask is on the nf-core Slack [#crisprseq](https://nfcore.slack.com/channels/crisprseq) channel ([join our Slack here](https://nf-co.re/join/slack)). +:::info +If you need help using or modifying nf-core/crisprseq then the best place to ask is on the nf-core Slack [#crisprseq](https://nfcore.slack.com/channels/crisprseq) channel ([join our Slack here](https://nf-co.re/join/slack)). +::: ## Contribution workflow diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml index 888cb4bc..b8bdd214 100644 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -78,7 +78,7 @@ jobs: - uses: actions/setup-python@v4 with: - python-version: "3.8" + python-version: "3.11" architecture: "x64" - name: Install dependencies diff --git a/.github/workflows/release-announcments.yml b/.github/workflows/release-announcments.yml new file mode 100644 index 00000000..6ad33927 --- /dev/null +++ b/.github/workflows/release-announcments.yml @@ -0,0 +1,68 @@ +name: release-announcements +# Automatic release toot and tweet anouncements +on: + release: + types: [published] + workflow_dispatch: + +jobs: + toot: + runs-on: ubuntu-latest + steps: + - uses: rzr/fediverse-action@master + with: + access-token: ${{ secrets.MASTODON_ACCESS_TOKEN }} + host: "mstdn.science" # custom host if not "mastodon.social" (default) + # GitHub event payload + # https://docs.github.com/en/developers/webhooks-and-events/webhooks/webhook-events-and-payloads#release + message: | + Pipeline release! ${{ github.repository }} v${{ github.event.release.tag_name }} - ${{ github.event.release.name }}! + + Please see the changelog: ${{ github.event.release.html_url }} + + send-tweet: + runs-on: ubuntu-latest + + steps: + - uses: actions/setup-python@v4 + with: + python-version: "3.10" + - name: Install dependencies + run: pip install tweepy==4.14.0 + - name: Send tweet + shell: python + run: | + import os + import tweepy + + client = tweepy.Client( + access_token=os.getenv("TWITTER_ACCESS_TOKEN"), + access_token_secret=os.getenv("TWITTER_ACCESS_TOKEN_SECRET"), + consumer_key=os.getenv("TWITTER_CONSUMER_KEY"), + consumer_secret=os.getenv("TWITTER_CONSUMER_SECRET"), + ) + tweet = os.getenv("TWEET") + client.create_tweet(text=tweet) + env: + TWEET: | + Pipeline release! ${{ github.repository }} v${{ github.event.release.tag_name }} - ${{ github.event.release.name }}! + + Please see the changelog: ${{ github.event.release.html_url }} + TWITTER_CONSUMER_KEY: ${{ secrets.TWITTER_CONSUMER_KEY }} + TWITTER_CONSUMER_SECRET: ${{ secrets.TWITTER_CONSUMER_SECRET }} + TWITTER_ACCESS_TOKEN: ${{ secrets.TWITTER_ACCESS_TOKEN }} + TWITTER_ACCESS_TOKEN_SECRET: ${{ secrets.TWITTER_ACCESS_TOKEN_SECRET }} + + bsky-post: + runs-on: ubuntu-latest + steps: + - uses: zentered/bluesky-post-action@v0.0.2 + with: + post: | + Pipeline release! ${{ github.repository }} v${{ github.event.release.tag_name }} - ${{ github.event.release.name }}! + + Please see the changelog: ${{ github.event.release.html_url }} + env: + BSKY_IDENTIFIER: ${{ secrets.BSKY_IDENTIFIER }} + BSKY_PASSWORD: ${{ secrets.BSKY_PASSWORD }} + # diff --git a/CHANGELOG.md b/CHANGELOG.md index 6f507b61..d48d4ecb 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ 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). -## v2.0.0 - [date] +## v2.1.0dev - [date] Initial release of nf-core/crisprseq, created with the [nf-core](https://nf-co.re/) template. diff --git a/CITATIONS.md b/CITATIONS.md index 287bcf20..9170aa49 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -12,7 +12,7 @@ - [FastQC](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) - > Andrews, S. (2010). FastQC: A Quality Control Tool for High Throughput Sequence Data [Online]. Available online https://www.bioinformatics.babraham.ac.uk/projects/fastqc/. + > Andrews, S. (2010). FastQC: A Quality Control Tool for High Throughput Sequence Data [Online]. - [MultiQC](https://pubmed.ncbi.nlm.nih.gov/27312411/) diff --git a/CODE_OF_CONDUCT.md b/CODE_OF_CONDUCT.md index f4fd052f..c089ec78 100644 --- a/CODE_OF_CONDUCT.md +++ b/CODE_OF_CONDUCT.md @@ -1,18 +1,20 @@ -# Code of Conduct at nf-core (v1.0) +# Code of Conduct at nf-core (v1.4) ## Our Pledge -In the interest of fostering an open, collaborative, and welcoming environment, we as contributors and maintainers of nf-core, pledge to making participation in our projects and community a harassment-free experience for everyone, regardless of: +In the interest of fostering an open, collaborative, and welcoming environment, we as contributors and maintainers of nf-core pledge to making participation in our projects and community a harassment-free experience for everyone, regardless of: - Age +- Ability - Body size +- Caste - Familial status - Gender identity and expression - Geographical location - Level of experience - Nationality and national origins - Native language -- Physical and neurological ability +- Neurodiversity - Race or ethnicity - Religion - Sexual identity and orientation @@ -22,80 +24,133 @@ Please note that the list above is alphabetised and is therefore not ranked in a ## Preamble -> Note: This Code of Conduct (CoC) has been drafted by the nf-core Safety Officer and been edited after input from members of the nf-core team and others. "We", in this document, refers to the Safety Officer and members of the nf-core core team, both of whom are deemed to be members of the nf-core community and are therefore required to abide by this Code of Conduct. This document will amended periodically to keep it up-to-date, and in case of any dispute, the most current version will apply. +:::note +This Code of Conduct (CoC) has been drafted by Renuka Kudva, Cris Tuñí, and Michael Heuer, with input from the nf-core Core Team and Susanna Marquez from the nf-core community. "We", in this document, refers to the Safety Officers and members of the nf-core Core Team, both of whom are deemed to be members of the nf-core community and are therefore required to abide by this Code of Conduct. This document will be amended periodically to keep it up-to-date. In case of any dispute, the most current version will apply. +::: -An up-to-date list of members of the nf-core core team can be found [here](https://nf-co.re/about). Our current safety officer is Renuka Kudva. +An up-to-date list of members of the nf-core core team can be found [here](https://nf-co.re/about). + +Our Safety Officers are Saba Nafees, Cris Tuñí, and Michael Heuer. nf-core is a young and growing community that welcomes contributions from anyone with a shared vision for [Open Science Policies](https://www.fosteropenscience.eu/taxonomy/term/8). Open science policies encompass inclusive behaviours and we strive to build and maintain a safe and inclusive environment for all individuals. -We have therefore adopted this code of conduct (CoC), which we require all members of our community and attendees in nf-core events to adhere to in all our workspaces at all times. Workspaces include but are not limited to Slack, meetings on Zoom, Jitsi, YouTube live etc. +We have therefore adopted this CoC, which we require all members of our community and attendees of nf-core events to adhere to in all our workspaces at all times. Workspaces include, but are not limited to, Slack, meetings on Zoom, gather.town, YouTube live etc. -Our CoC will be strictly enforced and the nf-core team reserve the right to exclude participants who do not comply with our guidelines from our workspaces and future nf-core activities. +Our CoC will be strictly enforced and the nf-core team reserves the right to exclude participants who do not comply with our guidelines from our workspaces and future nf-core activities. -We ask all members of our community to help maintain a supportive and productive workspace and to avoid behaviours that can make individuals feel unsafe or unwelcome. Please help us maintain and uphold this CoC. +We ask all members of our community to help maintain supportive and productive workspaces and to avoid behaviours that can make individuals feel unsafe or unwelcome. Please help us maintain and uphold this CoC. -Questions, concerns or ideas on what we can include? Contact safety [at] nf-co [dot] re +Questions, concerns, or ideas on what we can include? Contact members of the Safety Team on Slack or email safety [at] nf-co [dot] re. ## Our Responsibilities -The safety officer is responsible for clarifying the standards of acceptable behavior and are expected to take appropriate and fair corrective action in response to any instances of unacceptable behaviour. +Members of the Safety Team (the Safety Officers) are responsible for clarifying the standards of acceptable behavior and are expected to take appropriate and fair corrective action in response to any instances of unacceptable behaviour. -The safety officer in consultation with the nf-core core team have the right and responsibility to remove, edit, or reject comments, commits, code, wiki edits, issues, and other contributions that are not aligned to this Code of Conduct, or to ban temporarily or permanently any contributor for other behaviors that they deem inappropriate, threatening, offensive, or harmful. +The Safety Team, in consultation with the nf-core core team, have the right and responsibility to remove, edit, or reject comments, commits, code, wiki edits, issues, and other contributions that are not aligned to this CoC, or to ban temporarily or permanently any contributor for other behaviors that they deem inappropriate, threatening, offensive, or harmful. -Members of the core team or the safety officer who violate the CoC will be required to recuse themselves pending investigation. They will not have access to any reports of the violations and be subject to the same actions as others in violation of the CoC. +Members of the core team or the Safety Team who violate the CoC will be required to recuse themselves pending investigation. They will not have access to any reports of the violations and will be subject to the same actions as others in violation of the CoC. -## When are where does this Code of Conduct apply? +## When and where does this Code of Conduct apply? -Participation in the nf-core community is contingent on following these guidelines in all our workspaces and events. This includes but is not limited to the following listed alphabetically and therefore in no order of preference: +Participation in the nf-core community is contingent on following these guidelines in all our workspaces and events, such as hackathons, workshops, bytesize, and collaborative workspaces on gather.town. These guidelines include, but are not limited to, the following (listed alphabetically and therefore in no order of preference): - Communicating with an official project email address. - Communicating with community members within the nf-core Slack channel. - Participating in hackathons organised by nf-core (both online and in-person events). -- Participating in collaborative work on GitHub, Google Suite, community calls, mentorship meetings, email correspondence. -- Participating in workshops, training, and seminar series organised by nf-core (both online and in-person events). This applies to events hosted on web-based platforms such as Zoom, Jitsi, YouTube live etc. +- Participating in collaborative work on GitHub, Google Suite, community calls, mentorship meetings, email correspondence, and on the nf-core gather.town workspace. +- Participating in workshops, training, and seminar series organised by nf-core (both online and in-person events). This applies to events hosted on web-based platforms such as Zoom, gather.town, Jitsi, YouTube live etc. - Representing nf-core on social media. This includes both official and personal accounts. ## nf-core cares 😊 -nf-core's CoC and expectations of respectful behaviours for all participants (including organisers and the nf-core team) include but are not limited to the following (listed in alphabetical order): +nf-core's CoC and expectations of respectful behaviours for all participants (including organisers and the nf-core team) include, but are not limited to, the following (listed in alphabetical order): - Ask for consent before sharing another community member’s personal information (including photographs) on social media. - Be respectful of differing viewpoints and experiences. We are all here to learn from one another and a difference in opinion can present a good learning opportunity. -- Celebrate your accomplishments at events! (Get creative with your use of emojis 🎉 🥳 💯 🙌 !) +- Celebrate your accomplishments! (Get creative with your use of emojis 🎉 🥳 💯 🙌 !) - Demonstrate empathy towards other community members. (We don’t all have the same amount of time to dedicate to nf-core. If tasks are pending, don’t hesitate to gently remind members of your team. If you are leading a task, ask for help if you feel overwhelmed.) - Engage with and enquire after others. (This is especially important given the geographically remote nature of the nf-core community, so let’s do this the best we can) - Focus on what is best for the team and the community. (When in doubt, ask) -- Graciously accept constructive criticism, yet be unafraid to question, deliberate, and learn. +- Accept feedback, yet be unafraid to question, deliberate, and learn. - Introduce yourself to members of the community. (We’ve all been outsiders and we know that talking to strangers can be hard for some, but remember we’re interested in getting to know you and your visions for open science!) -- Show appreciation and **provide clear feedback**. (This is especially important because we don’t see each other in person and it can be harder to interpret subtleties. Also remember that not everyone understands a certain language to the same extent as you do, so **be clear in your communications to be kind.**) +- Show appreciation and **provide clear feedback**. (This is especially important because we don’t see each other in person and it can be harder to interpret subtleties. Also remember that not everyone understands a certain language to the same extent as you do, so **be clear in your communication to be kind.**) - Take breaks when you feel like you need them. -- Using welcoming and inclusive language. (Participants are encouraged to display their chosen pronouns on Zoom or in communication on Slack.) +- Use welcoming and inclusive language. (Participants are encouraged to display their chosen pronouns on Zoom or in communication on Slack) ## nf-core frowns on 😕 -The following behaviours from any participants within the nf-core community (including the organisers) will be considered unacceptable under this code of conduct. Engaging or advocating for any of the following could result in expulsion from nf-core workspaces. +The following behaviours from any participants within the nf-core community (including the organisers) will be considered unacceptable under this CoC. Engaging or advocating for any of the following could result in expulsion from nf-core workspaces: - Deliberate intimidation, stalking or following and sustained disruption of communication among participants of the community. This includes hijacking shared screens through actions such as using the annotate tool in conferencing software such as Zoom. - “Doxing” i.e. posting (or threatening to post) another person’s personal identifying information online. - Spamming or trolling of individuals on social media. -- Use of sexual or discriminatory imagery, comments, or jokes and unwelcome sexual attention. -- Verbal and text comments that reinforce social structures of domination related to gender, gender identity and expression, sexual orientation, ability, physical appearance, body size, race, age, religion or work experience. +- Use of sexual or discriminatory imagery, comments, jokes, or unwelcome sexual attention. +- Verbal and text comments that reinforce social structures of domination related to gender, gender identity and expression, sexual orientation, ability, physical appearance, body size, race, age, religion, or work experience. ### Online Trolling -The majority of nf-core interactions and events are held online. Unfortunately, holding events online comes with the added issue of online trolling. This is unacceptable, reports of such behaviour will be taken very seriously, and perpetrators will be excluded from activities immediately. +The majority of nf-core interactions and events are held online. Unfortunately, holding events online comes with the risk of online trolling. This is unacceptable — reports of such behaviour will be taken very seriously and perpetrators will be excluded from activities immediately. -All community members are required to ask members of the group they are working within for explicit consent prior to taking screenshots of individuals during video calls. +All community members are **required** to ask members of the group they are working with for explicit consent prior to taking screenshots of individuals during video calls. -## Procedures for Reporting CoC violations +## Procedures for reporting CoC violations If someone makes you feel uncomfortable through their behaviours or actions, report it as soon as possible. -You can reach out to members of the [nf-core core team](https://nf-co.re/about) and they will forward your concerns to the safety officer(s). +You can reach out to members of the Safety Team (Saba Nafees, Cris Tuñí, and Michael Heuer) on Slack. Alternatively, contact a member of the nf-core core team [nf-core core team](https://nf-co.re/about), and they will forward your concerns to the Safety Team. + +Issues directly concerning members of the Core Team or the Safety Team will be dealt with by other members of the core team and the safety manager — possible conflicts of interest will be taken into account. nf-core is also in discussions about having an ombudsperson and details will be shared in due course. + +All reports will be handled with the utmost discretion and confidentiality. + +You can also report any CoC violations to safety [at] nf-co [dot] re. In your email report, please do your best to include: + +- Your contact information. +- Identifying information (e.g. names, nicknames, pseudonyms) of the participant who has violated the Code of Conduct. +- The behaviour that was in violation and the circumstances surrounding the incident. +- The approximate time of the behaviour (if different than the time the report was made). +- Other people involved in the incident, if applicable. +- If you believe the incident is ongoing. +- If there is a publicly available record (e.g. mailing list record, a screenshot). +- Any additional information. + +After you file a report, one or more members of our Safety Team will contact you to follow up on your report. + +## Who will read and handle reports + +All reports will be read and handled by the members of the Safety Team at nf-core. + +If members of the Safety Team are deemed to have a conflict of interest with a report, they will be required to recuse themselves as per our Code of Conduct and will not have access to any follow-ups. + +To keep this first report confidential from any of the Safety Team members, please submit your first report by direct messaging on Slack/direct email to any of the nf-core members you are comfortable disclosing the information to, and be explicit about which member(s) you do not consent to sharing the information with. + +## Reviewing reports + +After receiving the report, members of the Safety Team will review the incident report to determine whether immediate action is required, for example, whether there is immediate threat to participants’ safety. + +The Safety Team, in consultation with members of the nf-core core team, will assess the information to determine whether the report constitutes a Code of Conduct violation, for them to decide on a course of action. + +In the case of insufficient information, one or more members of the Safety Team may contact the reporter, the reportee, or any other attendees to obtain more information. -Issues directly concerning members of the core team will be dealt with by other members of the core team and the safety manager, and possible conflicts of interest will be taken into account. nf-core is also in discussions about having an ombudsperson, and details will be shared in due course. +Once additional information is gathered, the Safety Team will collectively review and decide on the best course of action to take, if any. The Safety Team reserves the right to not act on a report. -All reports will be handled with utmost discretion and confidentially. +## Confidentiality + +All reports, and any additional information included, are only shared with the team of safety officers (and possibly members of the core team, in case the safety officer is in violation of the CoC). We will respect confidentiality requests for the purpose of protecting victims of abuse. + +We will not name harassment victims, beyond discussions between the safety officer and members of the nf-core team, without the explicit consent of the individuals involved. + +## Enforcement + +Actions taken by the nf-core’s Safety Team may include, but are not limited to: + +- Asking anyone to stop a behaviour. +- Asking anyone to leave the event and online spaces either temporarily, for the remainder of the event, or permanently. +- Removing access to the gather.town and Slack, either temporarily or permanently. +- Communicating to all participants to reinforce our expectations for conduct and remind what is unacceptable behaviour; this may be public for practical reasons. +- Communicating to all participants that an incident has taken place and how we will act or have acted — this may be for the purpose of letting event participants know we are aware of and dealing with the incident. +- Banning anyone from participating in nf-core-managed spaces, future events, and activities, either temporarily or permanently. +- No action. ## Attribution and Acknowledgements @@ -106,6 +161,22 @@ All reports will be handled with utmost discretion and confidentially. ## Changelog -### v1.0 - March 12th, 2021 +### v1.4 - February 8th, 2022 + +- Included a new member of the Safety Team. Corrected a typographical error in the text. + +### v1.3 - December 10th, 2021 + +- Added a statement that the CoC applies to nf-core gather.town workspaces. Corrected typographical errors in the text. + +### v1.2 - November 12th, 2021 + +- Removed information specific to reporting CoC violations at the Hackathon in October 2021. + +### v1.1 - October 14th, 2021 + +- Updated with names of new Safety Officers and specific information for the hackathon in October 2021. + +### v1.0 - March 15th, 2021 - Complete rewrite from original [Contributor Covenant](http://contributor-covenant.org/) CoC. diff --git a/LICENSE b/LICENSE index 41427d4d..4de6f708 100644 --- a/LICENSE +++ b/LICENSE @@ -1,6 +1,6 @@ MIT License -Copyright (c) mirpedrol +Copyright (c) Júlia Mir Pedrol, Laurence Kuhlburger 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 23db4ee3..d10e9a88 100644 --- a/README.md +++ b/README.md @@ -1,6 +1,7 @@ # ![nf-core/crisprseq](docs/images/nf-core-crisprseq_logo_light.png#gh-light-mode-only) ![nf-core/crisprseq](docs/images/nf-core-crisprseq_logo_dark.png#gh-dark-mode-only) -[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/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/crisprseq/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/crisprseq/actions?query=workflow%3A%22nf-core+CI%22) +[![GitHub Actions Linting Status](https://github.com/nf-core/crisprseq/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/crisprseq/actions?query=workflow%3A%22nf-core+linting%22)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) [![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A523.04.0-23aa62.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/) @@ -29,10 +30,11 @@ ## Usage -> **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. +:::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. +::: +- [@LaurenceKuhl](https://github.com/LaurenceKuhl) +- [@mirpedrol](https://github.com/mirpedrol) + +We thank the following people for their extensive assistance in the development of this pipeline: + +- [@ggabernet](https://github.com/ggabernet) +- [@jianhong](https://github.com/jianhong) +- [@mashehu](https://github.com/mashehu) +- [@msanvicente](https://github.com/msanvicente) +- [@SusiJo](https://github.com/SusiJo) ## Contributions and Support From d5c59ab89489c42ffe3727af3e5cb0497a677cde Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 7 Nov 2023 13:06:50 +0100 Subject: [PATCH 095/133] run black --- bin/BAGEL.py | 350 +++++++++++++++++++++++++++------------------------ 1 file changed, 189 insertions(+), 161 deletions(-) diff --git a/bin/BAGEL.py b/bin/BAGEL.py index 94bd734b..205fbb44 100755 --- a/bin/BAGEL.py +++ b/bin/BAGEL.py @@ -37,22 +37,21 @@ """ +import sys +import time import click import numpy as np import pandas as pd import scipy.stats as stats from sklearn.linear_model import LinearRegression -import sys -import time VERSION = 2.0 BUILD = 115 - -''' +""" Update history Build 115 @@ -77,26 +76,23 @@ 2. Now user can input column names 3. Fix Threshold function -''' +""" class OptionRequiredIf(click.Option): - def full_process_value(self, ctx, value): value = super(OptionRequiredIf, self).full_process_value(ctx, value) - if value is None and ctx.params['filter_multi_target'] is True: - msg = 'Error! Multi-target-filtering selected and not align-info provided.\n' \ - 'Please indicate align-info file.' + if value is None and ctx.params["filter_multi_target"] is True: + msg = ( + "Error! Multi-target-filtering selected and not align-info provided.\n" + "Please indicate align-info file." + ) raise click.MissingParameter(ctx=ctx, param=self, message=msg) return value - - - - -CONTEXT_SETTINGS = dict(help_option_names=['-h', '--help']) +CONTEXT_SETTINGS = dict(help_option_names=["-h", "--help"]) def round_to_hundredth(x): @@ -109,7 +105,6 @@ def func_linear(x, a, b): class Training: def __init__(self, X, n=None, cvnum=10): - if n == None: self._n = len(X) self._cvnum = cvnum @@ -118,7 +113,6 @@ def __init__(self, X, n=None, cvnum=10): self._X = X self._step = 0 - def cross_validation(self): if self._bid < 1: # bid check print("The number of genes is too small! n<" + str(self._cvnum)) @@ -157,7 +151,7 @@ def get_data(self, method=0): elif method == 1: train, test = self.cross_validation() else: - print('Method passed as a value that was neither 0 nor 1.') + print("Method passed as a value that was neither 0 nor 1.") sys.exit(1) return train, test @@ -208,24 +202,24 @@ def bagel(): """ -@click.command(name='version') +@click.command(name="version") def report_bagel_version(): """ Report the current build and version no. """ print( - 'Bayesian Analysis of Gene EssentiaLity (BAGEL) suite:\n' - 'Version: {VERSION}\n' - 'Build: {BUILD}'.format(VERSION=VERSION, BUILD=BUILD) + "Bayesian Analysis of Gene EssentiaLity (BAGEL) suite:\n" + "Version: {VERSION}\n" + "Build: {BUILD}".format(VERSION=VERSION, BUILD=BUILD) ) -@click.command(name='fc') -@click.option('-i', '--read-count-file', required=True, type=click.Path(exists=True)) -@click.option('-o', '--output-label', required=True) -@click.option('-c', '--control-columns', required=True) -@click.option('-m', '--min-reads', type=int, default=0) -@click.option('-Np', '--pseudo-count', type=int, default=5) +@click.command(name="fc") +@click.option("-i", "--read-count-file", required=True, type=click.Path(exists=True)) +@click.option("-o", "--output-label", required=True) +@click.option("-c", "--control-columns", required=True) +@click.option("-m", "--min-reads", type=int, default=0) +@click.option("-Np", "--pseudo-count", type=int, default=5) def calculate_fold_change(read_count_file, output_label, control_columns, min_reads, pseudo_count): """ \b @@ -259,9 +253,9 @@ def calculate_fold_change(read_count_file, output_label, control_columns, min_re # Import raw read data, normalize, filter for T0 min read counts # # Output: [output label].foldchange # # ---------------------------------------------------------------- # - reads = pd.read_csv(read_count_file, sep='\t', index_col=0) + reads = pd.read_csv(read_count_file, sep="\t", index_col=0) - reads[reads.columns.values[0]].fillna('NO_GENE_NAME', inplace=True) + reads[reads.columns.values[0]].fillna("NO_GENE_NAME", inplace=True) reads.fillna(0, inplace=True) control_columns = control_columns.split(",") # @@ -277,7 +271,7 @@ def calculate_fold_change(read_count_file, output_label, control_columns, min_re ctrl_sum = reads[ctrl_labels].sum(axis=1) reads.drop(ctrl_labels, axis=1, inplace=True) - ctrl_label_new = ';'.join(ctrl_labels) + ctrl_label_new = ";".join(ctrl_labels) reads[ctrl_label_new] = ctrl_sum except: print(reads[ctrl_labels].sum(axis=1)) @@ -298,9 +292,10 @@ def calculate_fold_change(read_count_file, output_label, control_columns, min_re # sumReads = reads.iloc[:, list(range(1, numColumns))].sum(0) normed = pd.DataFrame(index=reads.index.values) - normed['GENE'] = reads.iloc[:, 0] # first column is gene name - normed = reads.iloc[:, list(range(1, numColumns))] / np.tile(sumReads, - [numClones, 1]) * 10000000 # normalize to 10M reads + normed["GENE"] = reads.iloc[:, 0] # first column is gene name + normed = ( + reads.iloc[:, list(range(1, numColumns))] / np.tile(sumReads, [numClones, 1]) * 10000000 + ) # normalize to 10M reads # # filter for minimum readcount @@ -312,8 +307,8 @@ def calculate_fold_change(read_count_file, output_label, control_columns, min_re # calculate fold change # foldchange = pd.DataFrame(index=normed.index.values) - foldchange.index.name = 'REAGENT_ID' - foldchange['GENE'] = reads.iloc[f, 0] # dataframe 'normed' has no GENE column + foldchange.index.name = "REAGENT_ID" + foldchange["GENE"] = reads.iloc[f, 0] # dataframe 'normed' has no GENE column for i in range(numColumns - 1): foldchange[normed.columns.values[i]] = np.log2( (normed.loc[:, normed.columns.values[i]]) / normed[ctrl_label_new] @@ -328,39 +323,57 @@ def calculate_fold_change(read_count_file, output_label, control_columns, min_re # write foldchange file # - foldchange_filename = output_label + '.foldchange' - foldchange.to_csv(foldchange_filename, sep='\t', float_format='%4.3f') - - normedreads_filename = output_label + '.normed_readcount' - normed.to_csv(normedreads_filename, sep='\t', float_format='%3.2f') - - -@click.command(name='bf') -@click.option('-i', '--fold-change', required=True, type=click.Path(exists=True)) -@click.option('-o', '--output-file', required=True) -@click.option('-e', '--essential-genes', required=True, type=click.Path(exists=True)) -@click.option('-n', '--non-essential-genes', required=True, type=click.Path(exists=True)) -@click.option('-c', '--columns-to-test', required=True) -@click.option('-w', '--network-file', metavar='[network File]', default=None, type=click.Path(exists=True)) -@click.option('-m', '--filter-multi-target', is_flag=True) -@click.option('-m0', '--loci-without-mismatch', type=int, default=10) -@click.option('-m1', '--loci-with-mismatch', type=int, default=10) -@click.option('--align-info', metavar='--align-info [File]', default=None, - type=click.Path(exists=True), cls=OptionRequiredIf) -@click.option('-b', '--use-bootstrapping', is_flag=True) -@click.option('-NS', '--no-resampling', is_flag=True) -@click.option('-s', '--use-small-sample', is_flag=True) -@click.option('-N', '--no-of-cross-validations', type=int, default=10) -@click.option('-NB', '--bootstrap-iterations', type=int, default=1000) -@click.option('-r', '--sgrna-bayes-factors', is_flag=True) -@click.option('-f', '--equalise-sgrna-no', type=int) -@click.option('-s', '--seed', default=int(time.time() * 100000 % 100000), type=int) -@click.option('-t', '--run-test-mode', is_flag=True) -@click.option('-p', '--equalise-rep-no', type=int) + foldchange_filename = output_label + ".foldchange" + foldchange.to_csv(foldchange_filename, sep="\t", float_format="%4.3f") + + normedreads_filename = output_label + ".normed_readcount" + normed.to_csv(normedreads_filename, sep="\t", float_format="%3.2f") + + +@click.command(name="bf") +@click.option("-i", "--fold-change", required=True, type=click.Path(exists=True)) +@click.option("-o", "--output-file", required=True) +@click.option("-e", "--essential-genes", required=True, type=click.Path(exists=True)) +@click.option("-n", "--non-essential-genes", required=True, type=click.Path(exists=True)) +@click.option("-c", "--columns-to-test", required=True) +@click.option("-w", "--network-file", metavar="[network File]", default=None, type=click.Path(exists=True)) +@click.option("-m", "--filter-multi-target", is_flag=True) +@click.option("-m0", "--loci-without-mismatch", type=int, default=10) +@click.option("-m1", "--loci-with-mismatch", type=int, default=10) +@click.option( + "--align-info", metavar="--align-info [File]", default=None, type=click.Path(exists=True), cls=OptionRequiredIf +) +@click.option("-b", "--use-bootstrapping", is_flag=True) +@click.option("-NS", "--no-resampling", is_flag=True) +@click.option("-s", "--use-small-sample", is_flag=True) +@click.option("-N", "--no-of-cross-validations", type=int, default=10) +@click.option("-NB", "--bootstrap-iterations", type=int, default=1000) +@click.option("-r", "--sgrna-bayes-factors", is_flag=True) +@click.option("-f", "--equalise-sgrna-no", type=int) +@click.option("-s", "--seed", default=int(time.time() * 100000 % 100000), type=int) +@click.option("-t", "--run-test-mode", is_flag=True) +@click.option("-p", "--equalise-rep-no", type=int) def calculate_bayes_factors( - fold_change, output_file, essential_genes, non_essential_genes, columns_to_test, network_file, align_info, - use_bootstrapping, no_resampling, use_small_sample, filter_multi_target, loci_without_mismatch, loci_with_mismatch, - bootstrap_iterations, no_of_cross_validations, sgrna_bayes_factors, equalise_sgrna_no, seed, run_test_mode, equalise_rep_no + fold_change, + output_file, + essential_genes, + non_essential_genes, + columns_to_test, + network_file, + align_info, + use_bootstrapping, + no_resampling, + use_small_sample, + filter_multi_target, + loci_without_mismatch, + loci_with_mismatch, + bootstrap_iterations, + no_of_cross_validations, + sgrna_bayes_factors, + equalise_sgrna_no, + seed, + run_test_mode, + equalise_rep_no, ): """ \b @@ -460,7 +473,6 @@ def calculate_bayes_factors( multi_targeting_sgrnas_info = dict() if filter_multi_target: - try: aligninfo = pd.read_csv(align_info, header=None, index_col=0, sep="\t").fillna("") for seqid in aligninfo.index: @@ -480,7 +492,10 @@ def calculate_bayes_factors( multi_targeting_sgrnas[seqid] = True elif perfectmatch > 1 or mismatch_1bp > 0: multi_targeting_sgrnas_info[seqid] = ( - perfectmatch, mismatch_1bp, perfectmatch_gene, mismatch_1bp_gene + perfectmatch, + mismatch_1bp, + perfectmatch_gene, + mismatch_1bp_gene, ) except: @@ -494,19 +509,20 @@ def calculate_bayes_factors( # rnatagset = set() with open(fold_change) as fin: - fieldname = fin.readline().rstrip().split('\t') + fieldname = fin.readline().rstrip().split("\t") # # DEFINE CONTROLS # - columns = columns_to_test.split(',') + columns = columns_to_test.split(",") try: try: column_list = list(map(int, columns)) column_labels = [fieldname[x + 1] for x in column_list] except ValueError: column_labels = columns - column_list = [x for x in range(len(fieldname) - 1) if - fieldname[x + 1] in column_labels] # +1 because of First column start 2 + column_list = [ + x for x in range(len(fieldname) - 1) if fieldname[x + 1] in column_labels + ] # +1 because of First column start 2 print("Using column: " + ", ".join(column_labels)) # print "Using column: " + ", ".join(map(str,column_list)) @@ -515,7 +531,7 @@ def calculate_bayes_factors( sys.exit(1) for line in fin: - fields = line.rstrip().split('\t') + fields = line.rstrip().split("\t") rnatag = fields[0] if filter_multi_target is True: # multitargeting sgrna filtering if rnatag in multi_targeting_sgrnas: @@ -547,7 +563,7 @@ def calculate_bayes_factors( with open(essential_genes) as fin: skip_header = fin.readline() for line in fin: - coreEss.append(line.rstrip().split('\t')[0]) + coreEss.append(line.rstrip().split("\t")[0]) coreEss = np.array(coreEss) print("Number of reference essentials: " + str(len(coreEss))) @@ -555,7 +571,7 @@ def calculate_bayes_factors( with open(non_essential_genes) as fin: skip_header = fin.readline() for line in fin: - nonEss.append(line.rstrip().split('\t')[0]) + nonEss.append(line.rstrip().split("\t")[0]) nonEss = np.array(nonEss) print("Number of reference nonessentials: " + str(len(nonEss))) @@ -569,7 +585,7 @@ def calculate_bayes_factors( edgecount = 0 with open(network_file) as fin: for line in fin: - linearray = line.rstrip().split('\t') # GeneA \t GeneB format + linearray = line.rstrip().split("\t") # GeneA \t GeneB format if linearray[0] in genes_array and linearray[1] in genes_array: for i in [0, 1]: if linearray[i] not in network: @@ -586,8 +602,7 @@ def calculate_bayes_factors( # Define foldchange dynamic threshold. logarithm decay. # Parameters are defined by regression (achilles data) 2**-7 was used in previous version. - FC_THRESH = 2 ** (-1.1535 * np.log(len(np.intersect1d(genes_array, - nonEss)) + 13.324) + 0.7728) + FC_THRESH = 2 ** (-1.1535 * np.log(len(np.intersect1d(genes_array, nonEss)) + 13.324) + 0.7728) bf = {} boostedbf = {} for g in genes_array: @@ -603,8 +618,9 @@ def calculate_bayes_factors( # declare training class # training_data = Training(setdiff1d(gene_idx,np.where(in1d(genes_array,coreEss))),cvnum=NUMCV) # declare training class (only for Gold-standard gene set) - training_data = Training(np.where(np.in1d(genes_array, np.union1d(coreEss, nonEss)))[0], - cvnum=no_of_cross_validations) + training_data = Training( + np.where(np.in1d(genes_array, np.union1d(coreEss, nonEss)))[0], cvnum=no_of_cross_validations + ) # all non-goldstandards all_non_gs = np.where(np.logical_not(np.in1d(genes_array, np.union1d(coreEss, nonEss))))[0] else: @@ -649,7 +665,8 @@ def calculate_bayes_factors( if run_test_mode: fp.write( - "%d\n%s\n%s\n" % (loop, ",".join(genes_array[gene_train_idx]), ",".join(genes_array[gene_test_idx]))) + "%d\n%s\n%s\n" % (loop, ",".join(genes_array[gene_train_idx]), ",".join(genes_array[gene_test_idx])) + ) train_ess = np.where(np.in1d(genes_array[gene_train_idx], coreEss))[0] train_non = np.where(np.in1d(genes_array[gene_train_idx], nonEss))[0] @@ -661,14 +678,16 @@ def calculate_bayes_factors( # # define ess_train: vector of observed fold changes of essential genes in training set # - ess_train_fc_list_of_lists = [fc[rnatag] for g in genes_array[gene_train_idx[train_ess]] for rnatag in - gene2rna[g]] + ess_train_fc_list_of_lists = [ + fc[rnatag] for g in genes_array[gene_train_idx[train_ess]] for rnatag in gene2rna[g] + ] ess_train_fc_flat_list = [obs for sublist in ess_train_fc_list_of_lists for obs in list(sublist.values())] # # define non_train vector of observed fold changes of nonessential genes in training set # - non_train_fc_list_of_lists = [fc[rnatag] for g in genes_array[gene_train_idx[train_non]] for rnatag in - gene2rna[g]] + non_train_fc_list_of_lists = [ + fc[rnatag] for g in genes_array[gene_train_idx[train_non]] for rnatag in gene2rna[g] + ] non_train_fc_flat_list = [obs for sublist in non_train_fc_list_of_lists for obs in list(sublist.values())] # # calculate empirical fold change distributions for both @@ -781,21 +800,24 @@ def calculate_bayes_factors( if len(onlytarget) > 0: # comparsion between sgRNAs targeting one locus and multiple loci if len(multitarget) > 0: - bf_only = np.mean([sum(list(bf_mean_rna_rep[seqid].values())) for seqid in onlytarget]) for seqid in onlytarget: trainset[seqid] = [1, 0, 0] for seqid in multitarget: - if multi_targeting_sgrnas_info[seqid][2] > 1 or multi_targeting_sgrnas_info[seqid][ - 3] > 0: # train model using multi-targeting only targeting one protein coding gene + if ( + multi_targeting_sgrnas_info[seqid][2] > 1 or multi_targeting_sgrnas_info[seqid][3] > 0 + ): # train model using multi-targeting only targeting one protein coding gene continue count += 1 increment = sum(list(bf_mean_rna_rep[seqid].values())) - bf_only - trainset[seqid] = [multi_targeting_sgrnas_info[seqid][0], multi_targeting_sgrnas_info[seqid][1], - increment] + trainset[seqid] = [ + multi_targeting_sgrnas_info[seqid][0], + multi_targeting_sgrnas_info[seqid][1], + increment, + ] if count < 10: print("Not enough train set for calculating multi-targeting effect.\n") @@ -804,35 +826,40 @@ def calculate_bayes_factors( filter_multi_target = False else: - trainset = pd.DataFrame().from_dict(trainset).T X = trainset[[0, 1]] y = trainset[2] regressor = LinearRegression() regressor.fit(X, y) - coeff_df = pd.DataFrame(regressor.coef_, X.columns, columns=['Coefficient']) + coeff_df = pd.DataFrame(regressor.coef_, X.columns, columns=["Coefficient"]) for i in [0, 1]: - if coeff_df['Coefficient'][i] < 0: + if coeff_df["Coefficient"][i] < 0: print("Regression coefficient is below than zero. Substituted to zero\n") - coeff_df['Coefficient'][i] = 0.0 - print("Multiple effects from perfect matched loci = %.3f and 1bp mis-matched loci = %.3f" % ( - coeff_df['Coefficient'][0], coeff_df['Coefficient'][1])) + coeff_df["Coefficient"][i] = 0.0 + print( + "Multiple effects from perfect matched loci = %.3f and 1bp mis-matched loci = %.3f" + % (coeff_df["Coefficient"][0], coeff_df["Coefficient"][1]) + ) if rna_level == False: for g in gene2rna: penalty = 0.0 for seqid in gene2rna[g]: if seqid in multi_targeting_sgrnas_info: - penalty += float(multi_targeting_sgrnas_info[seqid][0] - 1) * coeff_df['Coefficient'][ - 0] + float(multi_targeting_sgrnas_info[seqid][1]) * coeff_df['Coefficient'][1] + penalty += ( + float(multi_targeting_sgrnas_info[seqid][0] - 1) * coeff_df["Coefficient"][0] + + float(multi_targeting_sgrnas_info[seqid][1]) * coeff_df["Coefficient"][1] + ) bf_multi_corrected_gene[g] = bf_mean[g] - penalty else: for g in gene2rna: for seqid in gene2rna[g]: if seqid in multi_targeting_sgrnas_info: - penalty = float(multi_targeting_sgrnas_info[seqid][0] - 1) * coeff_df['Coefficient'][ - 0] + float(multi_targeting_sgrnas_info[seqid][1]) * coeff_df['Coefficient'][1] + penalty = ( + float(multi_targeting_sgrnas_info[seqid][0] - 1) * coeff_df["Coefficient"][0] + + float(multi_targeting_sgrnas_info[seqid][1]) * coeff_df["Coefficient"][1] + ) else: penalty = 0.0 bf_multi_corrected_rna[seqid] = sum(list(bf_mean_rna_rep[seqid].values())) - penalty @@ -850,12 +877,12 @@ def calculate_bayes_factors( multiple_factor = equalise_sgrna_no / float(len(gene2rna[g])) bf_norm[g] = targetbf[g] * multiple_factor - ''' + """ if bf_std[rnatag] == 0.0: bf_norm[rnatag] = float('inf') else: bf_norm[g] = ( bf[rnatag] - bf_mean[rnatag] ) / bf_std[rnatag] - ''' + """ training_data = Training(gene_idx) # set training class reset # @@ -961,102 +988,100 @@ def calculate_bayes_factors( if run_test_mode == True: fp.close() - - - # # print out results # # Equalizing factor (Replicates) - if flat_rep==True: - eqf = equalise_rep_no/float(len(column_labels)) + if flat_rep == True: + eqf = equalise_rep_no / float(len(column_labels)) else: eqf = 1 # print out - with open(output_file, 'w') as fout: - + with open(output_file, "w") as fout: if rna_level == True: - fout.write('RNA\tGENE') + fout.write("RNA\tGENE") for i in range(len(column_list)): - fout.write('\t{0:s}'.format(column_labels[i])) + fout.write("\t{0:s}".format(column_labels[i])) if train_method == 0: - fout.write('\t{0:s}'.format(column_labels[i] + "_STD")) - fout.write('\tBF') + fout.write("\t{0:s}".format(column_labels[i] + "_STD")) + fout.write("\tBF") if train_method == 0: - fout.write('\tNumObs') - fout.write('\n') + fout.write("\tNumObs") + fout.write("\n") for rnatag in sorted(bf.keys()): # RNA tag - fout.write('{0:s}\t'.format(rnatag)) + fout.write("{0:s}\t".format(rnatag)) # Gene gene = rna2gene[rnatag] - fout.write('{0:s}\t'.format(gene)) + fout.write("{0:s}\t".format(gene)) # BF of replicates for rep in column_list: - fout.write('{0:4.3f}\t'.format(bf_mean_rna_rep[rnatag][rep])) + fout.write("{0:4.3f}\t".format(bf_mean_rna_rep[rnatag][rep])) if train_method == 0: - fout.write('{0:4.3f}\t'.format(bf_std_rna_rep[rnatag][rep])) + fout.write("{0:4.3f}\t".format(bf_std_rna_rep[rnatag][rep])) # Sum BF of replicates if filter_multi_target == True: - fout.write('{0:4.3f}'.format(float(bf_multi_corrected_rna[rnatag]) * eqf)) # eqf = equalizing factor for the number of replicates + fout.write( + "{0:4.3f}".format(float(bf_multi_corrected_rna[rnatag]) * eqf) + ) # eqf = equalizing factor for the number of replicates else: - fout.write('{0:4.3f}'.format(float(sum(list(bf_mean_rna_rep[rnatag].values()))) * eqf)) + fout.write("{0:4.3f}".format(float(sum(list(bf_mean_rna_rep[rnatag].values()))) * eqf)) # Num obs if train_method == 0: - fout.write('\t{0:d}'.format(num_obs[gene])) - fout.write('\n') + fout.write("\t{0:d}".format(num_obs[gene])) + fout.write("\n") else: - fout.write('GENE') + fout.write("GENE") if network_boost == True: - fout.write('\tBoostedBF') + fout.write("\tBoostedBF") if train_method == 0: - fout.write('\tSTD_BoostedBF') - fout.write('\tBF') + fout.write("\tSTD_BoostedBF") + fout.write("\tBF") if train_method == 0: - fout.write('\tSTD\tNumObs') + fout.write("\tSTD\tNumObs") if flat_sgrna == True: - fout.write('\tNormBF') - fout.write('\n') + fout.write("\tNormBF") + fout.write("\n") for g in sorted(genes.keys()): # Gene - fout.write('{0:s}'.format(g)) + fout.write("{0:s}".format(g)) if network_boost == True: boostedbf_mean = np.mean(boostedbf[g]) boostedbf_std = np.std(boostedbf[g]) - fout.write('\t{0:4.3f}'.format(float(boostedbf_mean) * eqf)) + fout.write("\t{0:4.3f}".format(float(boostedbf_mean) * eqf)) if train_method == 0: - fout.write('\t{0:4.3f}'.format(float(boostedbf_std) * eqf)) + fout.write("\t{0:4.3f}".format(float(boostedbf_std) * eqf)) # BF if filter_multi_target == True: - fout.write('\t{0:4.3f}'.format(float(bf_multi_corrected_gene[g]) * eqf)) # eqf = equalizing factor for the number of replicates + fout.write( + "\t{0:4.3f}".format(float(bf_multi_corrected_gene[g]) * eqf) + ) # eqf = equalizing factor for the number of replicates else: - fout.write('\t{0:4.3f}'.format(float(bf_mean[g]) * eqf )) + fout.write("\t{0:4.3f}".format(float(bf_mean[g]) * eqf)) # STD, Count if train_method == 0: - fout.write('\t{0:4.3f}\t{1:d}'.format(float(bf_std[g]), num_obs[g])) + fout.write("\t{0:4.3f}\t{1:d}".format(float(bf_std[g]), num_obs[g])) # Normalized BF if flat_sgrna == True: - fout.write('\t{0:4.3f}'.format(float(bf_norm[g]))) + fout.write("\t{0:4.3f}".format(float(bf_norm[g]))) - fout.write('\n') + fout.write("\n") -@click.command(name='pr') -@click.option('-i', '--bayes-factors', required=True, - type=click.Path(exists=True)) -@click.option('-o', '--output-file', required=True) -@click.option('-e', '--essential-genes', required=True, - type=click.Path(exists=True)) -@click.option('-n', '--non-essential-genes', required=True, type=click.Path(exists=True)) -@click.option('-k', '--use-column', default=None) +@click.command(name="pr") +@click.option("-i", "--bayes-factors", required=True, type=click.Path(exists=True)) +@click.option("-o", "--output-file", required=True) +@click.option("-e", "--essential-genes", required=True, type=click.Path(exists=True)) +@click.option("-n", "--non-essential-genes", required=True, type=click.Path(exists=True)) +@click.option("-k", "--use-column", default=None) def calculate_precision_recall(bayes_factors, output_file, essential_genes, non_essential_genes, use_column): """ Calculate precision-recall from an input Bayes Factors file: @@ -1093,38 +1118,41 @@ def calculate_precision_recall(bayes_factors, output_file, essential_genes, non_ print("Error! the column name is not in the file") sys.exit(1) else: - bf_column = 'BF' + bf_column = "BF" bf.sort_values(by=bf_column, ascending=False, inplace=True) - cumulative_tp = 0. - cumulative_fp = 0. - precision = 1. - recall = 0. + cumulative_tp = 0.0 + cumulative_fp = 0.0 + precision = 1.0 + recall = 0.0 # note float formats ess = essentials.index.values non = nonessentials.index.values totNumEssentials = len([x for x in bf.index.values if x in ess]) - with open(output_file, 'w') as fout: - - fout.write('Gene\t') + with open(output_file, "w") as fout: + fout.write("Gene\t") fout.write(bf_column) - fout.write('\tRecall\tPrecision\tFDR\n') + fout.write("\tRecall\tPrecision\tFDR\n") for g in bf.index.values: - if (g in ess): + if g in ess: cumulative_tp += 1 - elif (g in non): + elif g in non: cumulative_fp += 1 recall = cumulative_tp / totNumEssentials - if ((cumulative_tp > 0) | (cumulative_fp > 0)): + if (cumulative_tp > 0) | (cumulative_fp > 0): precision = cumulative_tp / (cumulative_tp + cumulative_fp) - fout.write('{0:s}\t{1:4.3f}\t{2:4.3f}\t{3:4.3f}\t{4:4.3f}\n'.format(g, bf.loc[g, bf_column], recall, precision, 1.0-precision)) + fout.write( + "{0:s}\t{1:4.3f}\t{2:4.3f}\t{3:4.3f}\t{4:4.3f}\n".format( + g, bf.loc[g, bf_column], recall, precision, 1.0 - precision + ) + ) -if __name__ == '__main__': +if __name__ == "__main__": bagel.add_command(calculate_fold_change) bagel.add_command(calculate_bayes_factors) bagel.add_command(calculate_precision_recall) From 49306903fd7bc5ca8751be3c9c7d6cb1dfce63d3 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 7 Nov 2023 13:22:47 +0100 Subject: [PATCH 096/133] fix left-padding spaces --- modules/local/mageck/graphrra.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/mageck/graphrra.nf b/modules/local/mageck/graphrra.nf index c4f77c6d..2c531d5d 100644 --- a/modules/local/mageck/graphrra.nf +++ b/modules/local/mageck/graphrra.nf @@ -43,8 +43,8 @@ process MAGECK_GRAPHRRA { gdata <- transform(gdata, Rank = rank(Score)) png(filename = paste0("$meta.treatment","_vs_","$meta.reference","_rank.png"), width = 6, height = 4, units = "in", res = 300) p1 = ScatterView(gdata, x = "Rank", y = "Score", label = "id", - top = 5, auto_cut_y = TRUE, ylab = "Log2FC", - groups = c("top", "bottom")) + top = 5, auto_cut_y = TRUE, ylab = "Log2FC", + groups = c("top", "bottom")) print(p1) dev.off() From da83776937e0bf9c77517d32ed699e476377fdbc Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 7 Nov 2023 15:17:50 +0100 Subject: [PATCH 097/133] Remove a comment --- README.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/README.md b/README.md index 9657b7c1..cd4ef567 100644 --- a/README.md +++ b/README.md @@ -93,6 +93,8 @@ SAMPLE1,SAMPLE1_R1.fastq.gz,SAMPLE1_R2.fastq.gz,control For more details on how to build a sample sheet, please refer to the [usage documentation](https://nf-co.re/crisprseq/usage) +--> + Now, you can run the pipeline using: ```bash From a276caeb036b61d869469a7d7fc1e0cac1bd6c7f Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 7 Nov 2023 15:41:47 +0100 Subject: [PATCH 098/133] Fix template issues --- README.md | 10 ++- docs/output.md | 66 +-------------- modules.json | 68 +++++++++++++++- workflows/crisprseq.nf | 135 ------------------------------- workflows/crisprseq_screening.nf | 1 + workflows/crisprseq_targeted.nf | 1 + 6 files changed, 79 insertions(+), 202 deletions(-) delete mode 100644 workflows/crisprseq.nf diff --git a/README.md b/README.md index cd4ef567..7fdfec34 100644 --- a/README.md +++ b/README.md @@ -70,8 +70,6 @@ to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/usage/i with `-profile test` before running the workflow on actual data. ::: - - Now, you can run the pipeline using: ```bash @@ -119,6 +115,12 @@ For more details about the output files and reports, please refer to the nf-core/crisprseq was originally written by Júlia Mir Pedrol, Laurence Kuhlburger. +nf-core/crisprseq targeted is based on [CRISPR-A](https://doi.org/10.1101/2022.09.02.506351) [[Sanvicente-García, et.al. (2023)](https://doi.org/10.1371/journal.pcbi.1011137)], originally written by Marta Sanvicente García at [Translational Synthetic Biology](https://synbio.upf.edu/) from [Universitat Pompeu Fabra](https://www.upf.edu/home). nf-core/crisprseq was originally written by Júlia Mir Pedrol, Laurence Kuhlburger. +It was re-written in Nextflow DSL2 and is primarily maintained by Júlia Mir Pedrol (@mirpedrol) at [Quantitative Biology Center (QBiC)](https://www.qbic.uni-tuebingen.de/) from [Universität Tübingen](https://uni-tuebingen.de/en/). + +nf-core/crisprseq screening was written and is primarly maintained by Laurence Kuhlburger (@LaurenceKuhl) at [Quantitative Biology Center (QBiC)](https://www.qbic.uni-tuebingen.de/) from [Universität Tübingen](https://uni-tuebingen.de/en/). + + diff --git a/docs/output.md b/docs/output.md index ffc58e89..21eb08b0 100644 --- a/docs/output.md +++ b/docs/output.md @@ -2,70 +2,12 @@ ## Introduction -This document describes the output produced by the pipeline. Most of the plots are taken from the MultiQC report, which summarises results at the end of the pipeline. +This document describes the output produced by the pipeline. This document describes the output produced by the pipeline. Most of the plots are taken from the MultiQC report, which summarises results at the end of the pipeline. -The directories listed below will be created in the results directory after the pipeline has finished. All paths are relative to the top-level results directory. - +Please refer to the respective Output documentation: The directories listed below will be created in the results directory after the pipeline has finished. All paths are relative to the top-level results directory. -## Pipeline overview -The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps: +- [Output targeted](output/targeted.md) +- [Output screening](output/screening.md) -- [FastQC](#fastqc) - Raw read QC -- [MultiQC](#multiqc) - Aggregate report describing results and QC from the whole pipeline -- [Pipeline information](#pipeline-information) - Report metrics generated during the workflow execution - -### FastQC - -
      -Output files - -- `fastqc/` - - `*_fastqc.html`: FastQC report containing quality metrics. - - `*_fastqc.zip`: Zip archive containing the FastQC report, tab-delimited data file and plot images. - -
      - -[FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) gives general quality metrics about your sequenced reads. It provides information about the quality score distribution across your reads, per base sequence content (%A/T/G/C), adapter contamination and overrepresented sequences. For further reading and documentation see the [FastQC help pages](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/). - -![MultiQC - FastQC sequence counts plot](images/mqc_fastqc_counts.png) - -![MultiQC - FastQC mean quality scores plot](images/mqc_fastqc_quality.png) - -![MultiQC - FastQC adapter content plot](images/mqc_fastqc_adapter.png) - -:::note -The FastQC plots displayed in the MultiQC report shows _untrimmed_ reads. They may contain adapter sequence and potentially regions with low quality. -::: - -### MultiQC - -
      -Output files - -- `multiqc/` - - `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser. - - `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline. - - `multiqc_plots/`: directory containing static images from the report in various formats. - -
      - -[MultiQC](http://multiqc.info) is a visualization tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available in the report data directory. - -Results generated by MultiQC collate pipeline QC from supported tools e.g. FastQC. The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see . - -### Pipeline information - -
      -Output files - -- `pipeline_info/` - - Reports generated by Nextflow: `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.dot`/`pipeline_dag.svg`. - - Reports generated by the pipeline: `pipeline_report.html`, `pipeline_report.txt` and `software_versions.yml`. The `pipeline_report*` files will only be present if the `--email` / `--email_on_fail` parameter's are used when running the pipeline. - - Reformatted samplesheet files used as input to the pipeline: `samplesheet.valid.csv`. - - Parameters used by the pipeline run: `params.json`. - -
      - -[Nextflow](https://www.nextflow.io/docs/latest/tracing.html) provides excellent functionality for generating various reports relevant to the running and execution of the pipeline. This will allow you to troubleshoot errors with the running of the pipeline, and also provide you with other information such as launch commands, run times and resource usage. diff --git a/modules.json b/modules.json index 647cc52f..e396008c 100644 --- a/modules.json +++ b/modules.json @@ -49,13 +49,79 @@ }, "fastqc": { "branch": "master", - "git_sha": "bd8092b67b5103bdd52e300f75889442275c3117", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"] + }, + "mageck/count": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"], + "patch": "modules/nf-core/mageck/count/mageck-count.diff" + }, + "mageck/mle": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"], + "patch": "modules/nf-core/mageck/mle/mageck-mle.diff" + }, + "mageck/test": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"], + "patch": "modules/nf-core/mageck/test/mageck-test.diff" + }, + "medaka": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"], + "patch": "modules/nf-core/medaka/medaka.diff" + }, + "minimap2/align": { + "branch": "master", + "git_sha": "603ecbd9f45300c9788f197d2a15a005685b4220", + "installed_by": ["modules"], + "patch": "modules/nf-core/minimap2/align/minimap2-align.diff" + }, + "minimap2/index": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", "installed_by": ["modules"] }, "multiqc": { "branch": "master", "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", "installed_by": ["modules"] + }, + "pear": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"] + }, + "racon": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"] + }, + "samtools/index": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"] + }, + "seqtk/seq": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"] + }, + "vsearch/cluster": { + "branch": "master", + "git_sha": "603ecbd9f45300c9788f197d2a15a005685b4220", + "installed_by": ["modules"], + "patch": "modules/nf-core/vsearch/cluster/vsearch-cluster.diff" + }, + "vsearch/sort": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"] } } }, diff --git a/workflows/crisprseq.nf b/workflows/crisprseq.nf deleted file mode 100644 index 1fec328e..00000000 --- a/workflows/crisprseq.nf +++ /dev/null @@ -1,135 +0,0 @@ -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - PRINT PARAMS SUMMARY -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - -include { paramsSummaryLog; paramsSummaryMap } from 'plugin/nf-validation' - -def logo = NfcoreTemplate.logo(workflow, params.monochrome_logs) -def citation = '\n' + WorkflowMain.citation(workflow) + '\n' -def summary_params = paramsSummaryMap(workflow) - -// Print parameter summary log to screen -log.info logo + paramsSummaryLog(workflow) + citation - -WorkflowCrisprseq.initialise(params, log) - -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - CONFIG FILES -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - -ch_multiqc_config = Channel.fromPath("$projectDir/assets/multiqc_config.yml", checkIfExists: true) -ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath( params.multiqc_config, checkIfExists: true ) : Channel.empty() -ch_multiqc_logo = params.multiqc_logo ? Channel.fromPath( params.multiqc_logo, checkIfExists: true ) : Channel.empty() -ch_multiqc_custom_methods_description = params.multiqc_methods_description ? file(params.multiqc_methods_description, checkIfExists: true) : file("$projectDir/assets/methods_description_template.yml", checkIfExists: true) - -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - IMPORT LOCAL MODULES/SUBWORKFLOWS -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - -// -// SUBWORKFLOW: Consisting of a mix of local and nf-core/modules -// -include { INPUT_CHECK } from '../subworkflows/local/input_check' - -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - IMPORT NF-CORE MODULES/SUBWORKFLOWS -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - -// -// MODULE: Installed directly from nf-core/modules -// -include { FASTQC } from '../modules/nf-core/fastqc/main' -include { MULTIQC } from '../modules/nf-core/multiqc/main' -include { CUSTOM_DUMPSOFTWAREVERSIONS } from '../modules/nf-core/custom/dumpsoftwareversions/main' - -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - RUN MAIN WORKFLOW -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - -// Info required for completion email and summary -def multiqc_report = [] - -workflow CRISPRSEQ { - - ch_versions = Channel.empty() - - // - // SUBWORKFLOW: Read in samplesheet, validate and stage input files - // - INPUT_CHECK ( - file(params.input) - ) - ch_versions = ch_versions.mix(INPUT_CHECK.out.versions) - // TODO: OPTIONAL, you can use nf-validation plugin to create an input channel from the samplesheet with Channel.fromSamplesheet("input") - // See the documentation https://nextflow-io.github.io/nf-validation/samplesheets/fromSamplesheet/ - // ! There is currently no tooling to help you write a sample sheet schema - - // - // MODULE: Run FastQC - // - FASTQC ( - INPUT_CHECK.out.reads - ) - ch_versions = ch_versions.mix(FASTQC.out.versions.first()) - - CUSTOM_DUMPSOFTWAREVERSIONS ( - ch_versions.unique().collectFile(name: 'collated_versions.yml') - ) - - // - // MODULE: MultiQC - // - workflow_summary = WorkflowCrisprseq.paramsSummaryMultiqc(workflow, summary_params) - ch_workflow_summary = Channel.value(workflow_summary) - - methods_description = WorkflowCrisprseq.methodsDescriptionText(workflow, ch_multiqc_custom_methods_description, params) - ch_methods_description = Channel.value(methods_description) - - ch_multiqc_files = Channel.empty() - ch_multiqc_files = ch_multiqc_files.mix(ch_workflow_summary.collectFile(name: 'workflow_summary_mqc.yaml')) - ch_multiqc_files = ch_multiqc_files.mix(ch_methods_description.collectFile(name: 'methods_description_mqc.yaml')) - ch_multiqc_files = ch_multiqc_files.mix(CUSTOM_DUMPSOFTWAREVERSIONS.out.mqc_yml.collect()) - ch_multiqc_files = ch_multiqc_files.mix(FASTQC.out.zip.collect{it[1]}.ifEmpty([])) - - MULTIQC ( - ch_multiqc_files.collect(), - ch_multiqc_config.toList(), - ch_multiqc_custom_config.toList(), - ch_multiqc_logo.toList() - ) - multiqc_report = MULTIQC.out.report.toList() -} - -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - COMPLETION EMAIL AND SUMMARY -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - -workflow.onComplete { - if (params.email || params.email_on_fail) { - NfcoreTemplate.email(workflow, params, summary_params, projectDir, log, multiqc_report) - } - NfcoreTemplate.dump_parameters(workflow, params) - NfcoreTemplate.summary(workflow, params, log) - if (params.hook_url) { - NfcoreTemplate.IM_notification(workflow, params, summary_params, projectDir, log) - } -} - -/* -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ - THE END -~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ -*/ - diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index ff2a61b5..a4be8125 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -252,6 +252,7 @@ workflow.onComplete { if (params.email || params.email_on_fail) { NfcoreTemplate.email(workflow, params, summary_params, projectDir, log, multiqc_report) } + NfcoreTemplate.dump_parameters(workflow, params) NfcoreTemplate.summary(workflow, params, log) if (params.hook_url) { NfcoreTemplate.adaptivecard(workflow, params, summary_params, projectDir, log) diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 8792469e..6c4044ba 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -787,6 +787,7 @@ workflow.onComplete { if (params.email || params.email_on_fail) { NfcoreTemplate.email(workflow, params, summary_params, projectDir, log, multiqc_report) } + NfcoreTemplate.dump_parameters(workflow, params) NfcoreTemplate.summary(workflow, params, log) if (params.hook_url) { NfcoreTemplate.IM_notification(workflow, params, summary_params, projectDir, log) From 631eae278b3a1a90780122f9da847c6abc390b72 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 7 Nov 2023 15:43:55 +0100 Subject: [PATCH 099/133] Update README.md MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- README.md | 1 - 1 file changed, 1 deletion(-) diff --git a/README.md b/README.md index 7fdfec34..832a0ec1 100644 --- a/README.md +++ b/README.md @@ -113,7 +113,6 @@ For more details about the output files and reports, please refer to the ## Credits -nf-core/crisprseq was originally written by Júlia Mir Pedrol, Laurence Kuhlburger. nf-core/crisprseq targeted is based on [CRISPR-A](https://doi.org/10.1101/2022.09.02.506351) [[Sanvicente-García, et.al. (2023)](https://doi.org/10.1371/journal.pcbi.1011137)], originally written by Marta Sanvicente García at [Translational Synthetic Biology](https://synbio.upf.edu/) from [Universitat Pompeu Fabra](https://www.upf.edu/home). nf-core/crisprseq was originally written by Júlia Mir Pedrol, Laurence Kuhlburger. It was re-written in Nextflow DSL2 and is primarily maintained by Júlia Mir Pedrol (@mirpedrol) at [Quantitative Biology Center (QBiC)](https://www.qbic.uni-tuebingen.de/) from [Universität Tübingen](https://uni-tuebingen.de/en/). From becdc6e256ffa85b73d9367be1f8df13aa40bb70 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 7 Nov 2023 15:44:03 +0100 Subject: [PATCH 100/133] Update docs/output.md MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 21eb08b0..1e115eee 100644 --- a/docs/output.md +++ b/docs/output.md @@ -8,6 +8,6 @@ This document describes the output produced by the pipeline. This document descr Please refer to the respective Output documentation: The directories listed below will be created in the results directory after the pipeline has finished. All paths are relative to the top-level results directory. -- [Output targeted](output/targeted.md) +- [Output targeted](output/targeted.md) - [Output screening](output/screening.md) From 6a87c2cf64e1746a7ef14596cf0be3af1f783ced Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 7 Nov 2023 15:44:14 +0100 Subject: [PATCH 101/133] Update docs/output.md MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- docs/output.md | 1 - 1 file changed, 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 1e115eee..5e44a3f0 100644 --- a/docs/output.md +++ b/docs/output.md @@ -5,7 +5,6 @@ This document describes the output produced by the pipeline. This document describes the output produced by the pipeline. Most of the plots are taken from the MultiQC report, which summarises results at the end of the pipeline. -Please refer to the respective Output documentation: The directories listed below will be created in the results directory after the pipeline has finished. All paths are relative to the top-level results directory. - [Output targeted](output/targeted.md) From 82b874c6e56367ff9af2896c03b6abebc383844a Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 7 Nov 2023 15:44:25 +0100 Subject: [PATCH 102/133] Update docs/output.md MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 5e44a3f0..ca0c8c42 100644 --- a/docs/output.md +++ b/docs/output.md @@ -2,7 +2,7 @@ ## Introduction -This document describes the output produced by the pipeline. This document describes the output produced by the pipeline. Most of the plots are taken from the MultiQC report, which summarises results at the end of the pipeline. +This document describes the output produced by the pipeline. From 14f28c6cc97f154d7f80b6ed61c9af8b7ecd742c Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Tue, 7 Nov 2023 14:45:35 +0000 Subject: [PATCH 103/133] [automated] Fix linting with Prettier --- README.md | 5 +---- docs/output.md | 4 ---- 2 files changed, 1 insertion(+), 8 deletions(-) diff --git a/README.md b/README.md index 832a0ec1..38a74435 100644 --- a/README.md +++ b/README.md @@ -70,7 +70,6 @@ to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/usage/i with `-profile test` before running the workflow on actual data. ::: - First, prepare a samplesheet with your input data that looks as follows: `samplesheet.csv`: @@ -113,13 +112,11 @@ For more details about the output files and reports, please refer to the ## Credits - -nf-core/crisprseq targeted is based on [CRISPR-A](https://doi.org/10.1101/2022.09.02.506351) [[Sanvicente-García, et.al. (2023)](https://doi.org/10.1371/journal.pcbi.1011137)], originally written by Marta Sanvicente García at [Translational Synthetic Biology](https://synbio.upf.edu/) from [Universitat Pompeu Fabra](https://www.upf.edu/home). nf-core/crisprseq was originally written by Júlia Mir Pedrol, Laurence Kuhlburger. +nf-core/crisprseq targeted is based on [CRISPR-A](https://doi.org/10.1101/2022.09.02.506351) [[Sanvicente-García, et.al. (2023)](https://doi.org/10.1371/journal.pcbi.1011137)], originally written by Marta Sanvicente García at [Translational Synthetic Biology](https://synbio.upf.edu/) from [Universitat Pompeu Fabra](https://www.upf.edu/home). nf-core/crisprseq was originally written by Júlia Mir Pedrol, Laurence Kuhlburger. It was re-written in Nextflow DSL2 and is primarily maintained by Júlia Mir Pedrol (@mirpedrol) at [Quantitative Biology Center (QBiC)](https://www.qbic.uni-tuebingen.de/) from [Universität Tübingen](https://uni-tuebingen.de/en/). nf-core/crisprseq screening was written and is primarly maintained by Laurence Kuhlburger (@LaurenceKuhl) at [Quantitative Biology Center (QBiC)](https://www.qbic.uni-tuebingen.de/) from [Universität Tübingen](https://uni-tuebingen.de/en/). - diff --git a/docs/output.md b/docs/output.md index ca0c8c42..f112182b 100644 --- a/docs/output.md +++ b/docs/output.md @@ -4,9 +4,5 @@ This document describes the output produced by the pipeline. - - - - [Output targeted](output/targeted.md) - [Output screening](output/screening.md) - From 7e16e87d9935772ddf377bbe04e6adc1febd4012 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 7 Nov 2023 15:54:59 +0100 Subject: [PATCH 104/133] Fix fastqc --- modules/nf-core/fastqc/main.nf | 6 +----- 1 file changed, 1 insertion(+), 5 deletions(-) diff --git a/modules/nf-core/fastqc/main.nf b/modules/nf-core/fastqc/main.nf index 249f9064..07d5e433 100644 --- a/modules/nf-core/fastqc/main.nf +++ b/modules/nf-core/fastqc/main.nf @@ -29,11 +29,7 @@ process FASTQC { printf "%s %s\\n" $rename_to | while read old_name new_name; do [ -f "\${new_name}" ] || ln -s \$old_name \$new_name done - - fastqc \\ - $args \\ - --threads $task.cpus \\ - $renamed_files + fastqc $args --threads $task.cpus $renamed_files cat <<-END_VERSIONS > versions.yml "${task.process}": From 5d83f6d9b9acffb0eef7fcd48217289a4f2efa8e Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 7 Nov 2023 15:57:19 +0100 Subject: [PATCH 105/133] Update README.md MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 38a74435..803e70ed 100644 --- a/README.md +++ b/README.md @@ -112,7 +112,7 @@ For more details about the output files and reports, please refer to the ## Credits -nf-core/crisprseq targeted is based on [CRISPR-A](https://doi.org/10.1101/2022.09.02.506351) [[Sanvicente-García, et.al. (2023)](https://doi.org/10.1371/journal.pcbi.1011137)], originally written by Marta Sanvicente García at [Translational Synthetic Biology](https://synbio.upf.edu/) from [Universitat Pompeu Fabra](https://www.upf.edu/home). nf-core/crisprseq was originally written by Júlia Mir Pedrol, Laurence Kuhlburger. +nf-core/crisprseq targeted is based on [CRISPR-A](https://doi.org/10.1101/2022.09.02.506351) [[Sanvicente-García, et.al. (2023)](https://doi.org/10.1371/journal.pcbi.1011137)], originally written by Marta Sanvicente García at [Translational Synthetic Biology](https://synbio.upf.edu/) from [Universitat Pompeu Fabra](https://www.upf.edu/home). It was re-written in Nextflow DSL2 and is primarily maintained by Júlia Mir Pedrol (@mirpedrol) at [Quantitative Biology Center (QBiC)](https://www.qbic.uni-tuebingen.de/) from [Universität Tübingen](https://uni-tuebingen.de/en/). nf-core/crisprseq screening was written and is primarly maintained by Laurence Kuhlburger (@LaurenceKuhl) at [Quantitative Biology Center (QBiC)](https://www.qbic.uni-tuebingen.de/) from [Universität Tübingen](https://uni-tuebingen.de/en/). From cfe1fd82fe8ad5e08583c4cd300cfd27b68d9ad4 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 7 Nov 2023 15:57:28 +0100 Subject: [PATCH 106/133] Update docs/output.md MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- docs/output.md | 1 + 1 file changed, 1 insertion(+) diff --git a/docs/output.md b/docs/output.md index f112182b..f6e1cc39 100644 --- a/docs/output.md +++ b/docs/output.md @@ -3,6 +3,7 @@ ## Introduction This document describes the output produced by the pipeline. +Please refer to the respective Output documentation: - [Output targeted](output/targeted.md) - [Output screening](output/screening.md) From 5f191109e5a151913006d29877b125a794bcc5e6 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 7 Nov 2023 15:57:47 +0100 Subject: [PATCH 107/133] Update README.md MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit Co-authored-by: Júlia Mir Pedrol --- README.md | 1 + 1 file changed, 1 insertion(+) diff --git a/README.md b/README.md index 803e70ed..fbacd7bb 100644 --- a/README.md +++ b/README.md @@ -1,5 +1,6 @@ # ![nf-core/crisprseq](docs/images/nf-core-crisprseq_logo_light.png#gh-light-mode-only) ![nf-core/crisprseq](docs/images/nf-core-crisprseq_logo_dark.png#gh-dark-mode-only) +[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.8116072-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.8116072) [![GitHub Actions CI Status](https://github.com/nf-core/crisprseq/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/crisprseq/actions?query=workflow%3A%22nf-core+CI%22) [![GitHub Actions Linting Status](https://github.com/nf-core/crisprseq/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/crisprseq/actions?query=workflow%3A%22nf-core+linting%22)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX) From 8583f1ab8cb214fdc511df8bef4ca6a562cc8682 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 8 Nov 2023 09:26:02 +0100 Subject: [PATCH 108/133] Bump version --- CHANGELOG.md | 3 ++- assets/multiqc_config.yml | 4 ++-- nextflow.config | 2 +- 3 files changed, 5 insertions(+), 4 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 8fd0a0dc..3453b0ec 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ 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). -## [v2.1.0dev] +## [v2.1.0 - Margarita Salas](https://github.com/nf-core/crisprseq/releases/tag/2.1.0) - [08.11.2023] ### Added @@ -15,6 +15,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed - Change to `process_high` for the mageck mle module ([#60](https://github.com/nf-core/crisprseq/pull/60) +- Fix paired-end samplesheet file for screening ([#60](https://github.com/nf-core/crisprseq/pull/60) - Summary processes don't modify the input file anymore, allowing resuming these processes ([#66](https://github.com/nf-core/crisprseq/pull/66)) - Do not stash unexistent files, use empty lists instead. Fixes AWS tests ([#67](https://github.com/nf-core/crisprseq/pull/67)) - Rename process `merging_summary` to `preprocessing_summary` to improve clarity ([#69](https://github.com/nf-core/crisprseq/pull/69)) diff --git a/assets/multiqc_config.yml b/assets/multiqc_config.yml index 368215a8..a754e6ab 100644 --- a/assets/multiqc_config.yml +++ b/assets/multiqc_config.yml @@ -1,7 +1,7 @@ report_comment: > - This report has been generated by the       nf-core/crisprseq + This report has been generated by the nf-core/crisprseq analysis pipeline. For information about how to interpret these results, please see the - documentation. + documentation. report_section_order: "nf-core-crisprseq-methods-description": order: -1000 diff --git a/nextflow.config b/nextflow.config index adcec17c..1be05b5b 100644 --- a/nextflow.config +++ b/nextflow.config @@ -265,7 +265,7 @@ manifest { description = """Pipeline for the analysis of CRISPR data""" mainScript = 'main.nf' nextflowVersion = '!>=23.04.0' - version = '2.1.0dev' + version = '2.1.0' doi = '' } From 4bb1ef4c1fb7935764287b51d6e2ae15bf04c565 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Wed, 8 Nov 2023 11:30:19 +0100 Subject: [PATCH 109/133] Modify surname into a chips flavour --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 3453b0ec..c88307c2 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ 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). -## [v2.1.0 - Margarita Salas](https://github.com/nf-core/crisprseq/releases/tag/2.1.0) - [08.11.2023] +## [v2.1.0 - Jamon Salas](https://github.com/nf-core/crisprseq/releases/tag/2.1.0) - [08.11.2023] ### Added From 63000ada5044bf387dc409e4f86986e63c840e4c Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Thu, 9 Nov 2023 16:34:50 +0100 Subject: [PATCH 110/133] Add versions --- modules/local/bagel2/bf.nf | 7 ++++++- modules/local/bagel2/fc.nf | 12 ++++++++---- modules/local/bagel2/graph.nf | 12 +++++++++++- modules/local/bagel2/pr.nf | 9 +++++++-- modules/local/mageck/graphrra.nf | 15 +++++++++++++++ workflows/crisprseq_screening.nf | 19 ++++++++++++++++++- 6 files changed, 65 insertions(+), 9 deletions(-) diff --git a/modules/local/bagel2/bf.nf b/modules/local/bagel2/bf.nf index 9f5fe090..3ff86603 100644 --- a/modules/local/bagel2/bf.nf +++ b/modules/local/bagel2/bf.nf @@ -16,7 +16,7 @@ process BAGEL2_BF { output: tuple val(meta), path("*.bf"), emit: bf - //path "versions.yml" , emit: versions + path "versions.yml" , emit: versions when: task.ext.when == null || task.ext.when @@ -28,6 +28,11 @@ process BAGEL2_BF { """ BAGEL.py bf -i $foldchange -o '${meta.treatment}_vs_${meta.reference}.bf' $args -e $reference_essentials -n $reference_nonessentials -c ${meta.treatment} + cat <<-END_VERSIONS > versions.yml + "${task.process}": + python: \$(python --version | sed 's/Python //g') + BAGEL2: \$( BAGEL.py version | grep -o 'Version: [0-9.]*' | awk '{print \$2}' | grep -v '^\$') + END_VERSIONS """ } diff --git a/modules/local/bagel2/fc.nf b/modules/local/bagel2/fc.nf index bec12be1..b65afaea 100644 --- a/modules/local/bagel2/fc.nf +++ b/modules/local/bagel2/fc.nf @@ -11,9 +11,9 @@ process BAGEL2_FC { tuple val(meta), path(count_table) output: - tuple val(meta), path("*.foldchange"), emit: foldchange - tuple val(meta), path("*.normed_readcount"), emit: normed_counts - //path "versions.yml" , emit: versions + tuple val(meta), path("*.foldchange") , emit: foldchange + tuple val(meta), path("*.normed_readcount") , emit: normed_counts + path "versions.yml" , emit: versions when: task.ext.when == null || task.ext.when @@ -24,7 +24,11 @@ process BAGEL2_FC { """ BAGEL.py fc -i $count_table -o ${meta.treatment}_vs_${meta.reference} -c $meta.reference $args - + cat <<-END_VERSIONS > versions.yml + "${task.process}": + python: \$(python --version | sed 's/Python //g') + BAGEL2: \$( BAGEL.py version | grep -o 'Version: [0-9.]*' | awk '{print \$2}' | grep -v '^\$') + END_VERSIONS """ } diff --git a/modules/local/bagel2/graph.nf b/modules/local/bagel2/graph.nf index c14e7810..270c265f 100644 --- a/modules/local/bagel2/graph.nf +++ b/modules/local/bagel2/graph.nf @@ -13,7 +13,7 @@ process BAGEL2_GRAPH { output: path("*.png") , emit: pictures - + path "versions.yml" , emit: versions when: task.ext.when == null || task.ext.when @@ -60,6 +60,16 @@ process BAGEL2_GRAPH { plt.savefig(file_name) plt.show() + # Output version information + version = pd. __version__ + matplotlib_version = plt.matplotlib.__version__ + # alas, no `pyyaml` pre-installed in the cellranger container + with open("versions.yml", "w") as f: + f.write('"${task.process}":\\n') + f.write(f' pandas: "{version}"\\n') + f.write(f' matplotlib.pyplot: "{matplotlib_version}"\\n') + + """ diff --git a/modules/local/bagel2/pr.nf b/modules/local/bagel2/pr.nf index b5cd1793..cc4f3157 100644 --- a/modules/local/bagel2/pr.nf +++ b/modules/local/bagel2/pr.nf @@ -13,8 +13,7 @@ process BAGEL2_PR { output: tuple val(meta), path("*.pr") , emit: pr - - //path "versions.yml" , emit: versions + path "versions.yml" , emit: versions when: task.ext.when == null || task.ext.when @@ -25,6 +24,12 @@ process BAGEL2_PR { """ BAGEL.py pr -i $bf -o '${meta.treatment}_vs_${meta.reference}.pr' -e $reference_essentials -n $reference_nonessentials + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + python: \$(python --version | sed 's/Python //g') + BAGEL2: \$( BAGEL.py version | grep -o 'Version: [0-9.]*' | awk '{print \$2}' | grep -v '^\$') + END_VERSIONS """ } diff --git a/modules/local/mageck/graphrra.nf b/modules/local/mageck/graphrra.nf index 2c531d5d..ec11bf55 100644 --- a/modules/local/mageck/graphrra.nf +++ b/modules/local/mageck/graphrra.nf @@ -48,6 +48,21 @@ process MAGECK_GRAPHRRA { print(p1) dev.off() + version_file_path <- "versions.yml" + version_flute <- paste(unlist(packageVersion("MAGeCKFlute")), collapse = ".") + version_ggplot <- paste(unlist(packageVersion("MAGeCKFlute")), collapse = ".") + + f <- file(version_file_path, "w") + writeLines('"${task.process}":', f) + writeLines(" MAGeCKFlute: ", f, sep = "") + writeLines(version_flute, f) + writeLines(" ggplot2: ", f, sep = "") + writeLines(version_ggplot, f) + close(f) + + + + """ diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index ff2a61b5..95a9e22d 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -116,7 +116,7 @@ workflow CRISPRSEQ_SCREENING { [[id: condition, single_end: single_end], fastqs] } .set { joined } - joined.dump(tag:"metadata") + // // MODULE: Run mageck count @@ -147,6 +147,9 @@ workflow CRISPRSEQ_SCREENING { params.min_targeted_genes ) + ch_versions = ch_versions.mix(CRISPRCLEANR_NORMALIZE.out.versions) + + CRISPRCLEANR_NORMALIZE.out.norm_count_file.map { it -> it[1] }.set { ch_counts } @@ -162,9 +165,12 @@ workflow CRISPRSEQ_SCREENING { counts ) + ch_versions = ch_versions.mix(MAGECK_TEST.out.versions) + MAGECK_GRAPHRRA ( MAGECK_TEST.out.gene_summary ) + //TODO: Missing versions } if(params.rra_contrasts) { @@ -180,6 +186,7 @@ workflow CRISPRSEQ_SCREENING { BAGEL2_FC ( counts ) + ch_versions = ch_versions.mix(BAGEL2_FC.out.versions) BAGEL2_BF ( BAGEL2_FC.out.foldchange, @@ -187,12 +194,18 @@ workflow CRISPRSEQ_SCREENING { ch_bagel_reference_nonessentials ) + ch_versions = ch_versions.mix(BAGEL2_BF.out.versions) + + ch_bagel_pr = BAGEL2_BF.out.bf.combine(ch_bagel_reference_essentials) .combine(ch_bagel_reference_nonessentials) BAGEL2_PR ( ch_bagel_pr ) + + ch_versions = ch_versions.mix(BAGEL2_PR.out.versions) + BAGEL2_GRAPH ( BAGEL2_PR.out.pr ) @@ -207,6 +220,10 @@ workflow CRISPRSEQ_SCREENING { MAGECK_MLE ( ch_designed_mle ) + + ch_versions = ch_versions.mix(MAGECK_MLE.out.versions) + + } CUSTOM_DUMPSOFTWAREVERSIONS ( From c0f0e7455b710536d8cc153428893bc4ab3d0e31 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Thu, 9 Nov 2023 16:38:06 +0100 Subject: [PATCH 111/133] fix zenodo links citing all versions --- README.md | 6 +++--- lib/WorkflowMain.groovy | 2 +- 2 files changed, 4 insertions(+), 4 deletions(-) diff --git a/README.md b/README.md index 09ff2465..ffc72899 100644 --- a/README.md +++ b/README.md @@ -1,8 +1,8 @@ # ![nf-core/crisprseq](docs/images/nf-core-crisprseq_logo_light.png#gh-light-mode-only) ![nf-core/crisprseq](docs/images/nf-core-crisprseq_logo_dark.png#gh-dark-mode-only) -[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.8116072-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.8116072) +[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.7598496-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.7598496) [![GitHub Actions CI Status](https://github.com/nf-core/crisprseq/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/crisprseq/actions?query=workflow%3A%22nf-core+CI%22) -[![GitHub Actions Linting Status](https://github.com/nf-core/crisprseq/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/crisprseq/actions?query=workflow%3A%22nf-core+linting%22)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/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/crisprseq/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/crisprseq/actions?query=workflow%3A%22nf-core+linting%22)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results) [![Nextflow](https://img.shields.io/badge/nextflow%20DSL2-%E2%89%A523.04.0-23aa62.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/) @@ -139,7 +139,7 @@ For further information or help, don't hesitate to get in touch on the [Slack `# ## Citations -If you use nf-core/crisprseq for your analysis, please cite it using the following doi: [10.5281/zenodo.8116072](https://doi.org/10.5281/zenodo.8116072) +If you use nf-core/crisprseq for your analysis, please cite it using the following doi: [10.5281/zenodo.7598496](https://doi.org/10.5281/zenodo.7598496) An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file. diff --git a/lib/WorkflowMain.groovy b/lib/WorkflowMain.groovy index 36db450c..0e40058b 100755 --- a/lib/WorkflowMain.groovy +++ b/lib/WorkflowMain.groovy @@ -12,7 +12,7 @@ class WorkflowMain { public static String citation(workflow) { return "If you use ${workflow.manifest.name} for your analysis please cite:\n\n" + "* The pipeline\n" + - " https://doi.org/10.5281/zenodo.8116072\n\n" + + " https://doi.org/10.5281/zenodo.7598496\n\n" + "* The nf-core framework\n" + " https://doi.org/10.1038/s41587-020-0439-x\n\n" + "* Software dependencies\n" + From d40255956fd9062a0ec4e858ef3984cac4560cd8 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Fri, 10 Nov 2023 08:57:32 +0100 Subject: [PATCH 112/133] add template updates to all aws fulltests --- .github/workflows/awsfulltest.yml | 2 +- .github/workflows/awsfulltest_screening.yml | 10 +++++++--- 2 files changed, 8 insertions(+), 4 deletions(-) diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml index fb0e299f..df8badba 100644 --- a/.github/workflows/awsfulltest.yml +++ b/.github/workflows/awsfulltest.yml @@ -25,7 +25,7 @@ jobs: parameters: | { "hook_url": "${{ secrets.MEGATESTS_ALERTS_SLACK_HOOK_URL }}", - "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/crisprseq/results-${{ github.sha }}" + "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/crisprseq/results-${{ github.sha }}/targeted_test" } profiles: test_full diff --git a/.github/workflows/awsfulltest_screening.yml b/.github/workflows/awsfulltest_screening.yml index df435c92..8c91e9cf 100644 --- a/.github/workflows/awsfulltest_screening.yml +++ b/.github/workflows/awsfulltest_screening.yml @@ -14,18 +14,22 @@ jobs: runs-on: ubuntu-latest steps: - name: Launch workflow via tower - uses: nf-core/tower-action@v3 + uses: seqeralabs/action-tower-launch@v2 with: workspace_id: ${{ secrets.TOWER_WORKSPACE_ID }} access_token: ${{ secrets.TOWER_ACCESS_TOKEN }} compute_env: ${{ secrets.TOWER_COMPUTE_ENV }} - workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/crisprseq/work-${{ github.sha }}/screening_test + revision: ${{ github.sha }} + workdir: s3://${{ secrets.AWS_S3_BUCKET }}/work/crisprseq/work-${{ github.sha }} parameters: | { + "hook_url": "${{ secrets.MEGATESTS_ALERTS_SLACK_HOOK_URL }}", "outdir": "s3://${{ secrets.AWS_S3_BUCKET }}/crisprseq/results-${{ github.sha }}/screening_test" } profiles: test_screening_full - uses: actions/upload-artifact@v3 with: name: Tower debug log file - path: tower_action_*.log + path: | + tower_action_*.log + tower_action_*.json From 516519d73a033bb04e59b541195275ad60109120 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Fri, 10 Nov 2023 09:55:23 +0100 Subject: [PATCH 113/133] fix dump_parameters function according to dev version of the nf-core template --- lib/NfcoreTemplate.groovy | 13 ++++++------- 1 file changed, 6 insertions(+), 7 deletions(-) diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy index 01b8653d..783c3fab 100755 --- a/lib/NfcoreTemplate.groovy +++ b/lib/NfcoreTemplate.groovy @@ -227,15 +227,14 @@ class NfcoreTemplate { // Dump pipeline parameters in a json file // public static void dump_parameters(workflow, params) { - def output_d = new File("${params.outdir}/pipeline_info/") - if (!output_d.exists()) { - output_d.mkdirs() - } - def timestamp = new java.util.Date().format( 'yyyy-MM-dd_HH-mm-ss') - def output_pf = new File(output_d, "params_${timestamp}.json") + def filename = "params_${timestamp}.json" + def temp_pf = new File(workflow.launchDir.toString(), ".${filename}") def jsonStr = JsonOutput.toJson(params) - output_pf.text = JsonOutput.prettyPrint(jsonStr) + temp_pf.text = JsonOutput.prettyPrint(jsonStr) + + FilesEx.copyTo(temp_pf.toPath(), "${params.outdir}/pipeline_info/params_${timestamp}.json") + temp_pf.delete() } // From 4ca08acce4003aeb0556a48b7e57a792810ca89c Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Fri, 10 Nov 2023 10:21:08 +0100 Subject: [PATCH 114/133] add NfcoreTemplate.groovy to .nf-core.yml to fix linting --- .nf-core.yml | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/.nf-core.yml b/.nf-core.yml index 065203c3..94c07a2e 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -1,4 +1,4 @@ repository_type: pipeline lint: - # We skip these linting as we have splitted tests between targeted and screening - files_exist: conf/test.config + files_unchanged: + - lib/NfcoreTemplate.groovy # Introduced a change ahead of the nf-core/tools release From eec96e66ac3bd5d213419d388e2f6eec2471d1b0 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Fri, 10 Nov 2023 10:49:45 +0100 Subject: [PATCH 115/133] copy NfcoreTemplate.groovy from nf-core template 2.11dev --- lib/NfcoreTemplate.groovy | 19 ++++++++++++------- 1 file changed, 12 insertions(+), 7 deletions(-) diff --git a/lib/NfcoreTemplate.groovy b/lib/NfcoreTemplate.groovy index 783c3fab..e248e4c3 100755 --- a/lib/NfcoreTemplate.groovy +++ b/lib/NfcoreTemplate.groovy @@ -4,6 +4,7 @@ import org.yaml.snakeyaml.Yaml import groovy.json.JsonOutput +import nextflow.extension.FilesEx class NfcoreTemplate { @@ -141,12 +142,14 @@ class NfcoreTemplate { try { if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } // Try to send HTML e-mail using sendmail + def sendmail_tf = new File(workflow.launchDir.toString(), ".sendmail_tmp.html") + sendmail_tf.withWriter { w -> w << sendmail_html } [ 'sendmail', '-t' ].execute() << sendmail_html log.info "-${colors.purple}[$workflow.manifest.name]${colors.green} Sent summary e-mail to $email_address (sendmail)-" } catch (all) { // Catch failures and try with plaintext def mail_cmd = [ 'mail', '-s', subject, '--content-type=text/html', email_address ] - if ( mqc_report.size() <= max_multiqc_email_size.toBytes() ) { + if ( mqc_report != null && mqc_report.size() <= max_multiqc_email_size.toBytes() ) { mail_cmd += [ '-A', mqc_report ] } mail_cmd.execute() << email_html @@ -155,14 +158,16 @@ class NfcoreTemplate { } // Write summary e-mail HTML to a file - def output_d = new File("${params.outdir}/pipeline_info/") - if (!output_d.exists()) { - output_d.mkdirs() - } - def output_hf = new File(output_d, "pipeline_report.html") + def output_hf = new File(workflow.launchDir.toString(), ".pipeline_report.html") output_hf.withWriter { w -> w << email_html } - def output_tf = new File(output_d, "pipeline_report.txt") + FilesEx.copyTo(output_hf.toPath(), "${params.outdir}/pipeline_info/pipeline_report.html"); + output_hf.delete() + + // Write summary e-mail TXT to a file + def output_tf = new File(workflow.launchDir.toString(), ".pipeline_report.txt") output_tf.withWriter { w -> w << email_txt } + FilesEx.copyTo(output_tf.toPath(), "${params.outdir}/pipeline_info/pipeline_report.txt"); + output_tf.delete() } // From b24e0ce13aa229278e82e29f1dc8a3a4973c4d10 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Fri, 10 Nov 2023 11:00:35 +0100 Subject: [PATCH 116/133] fix .nf-core.yml --- .nf-core.yml | 2 ++ 1 file changed, 2 insertions(+) diff --git a/.nf-core.yml b/.nf-core.yml index 94c07a2e..085dbd0a 100644 --- a/.nf-core.yml +++ b/.nf-core.yml @@ -1,4 +1,6 @@ repository_type: pipeline lint: + files_exist: + - conf/test.config # We skip these linting as we have splitted tests between targeted and screening files_unchanged: - lib/NfcoreTemplate.groovy # Introduced a change ahead of the nf-core/tools release From 06b98c4ee8af18dc6cd84f6b6ddce430d023aada Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C3=BAlia=20Mir=20Pedrol?= Date: Fri, 10 Nov 2023 11:20:35 +0100 Subject: [PATCH 117/133] Apply suggestions from code review --- CHANGELOG.md | 4 ++-- conf/test_screening.config | 2 +- nextflow.config | 2 +- 3 files changed, 4 insertions(+), 4 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index c88307c2..9f846e3b 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -14,8 +14,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 ### Fixed -- Change to `process_high` for the mageck mle module ([#60](https://github.com/nf-core/crisprseq/pull/60) -- Fix paired-end samplesheet file for screening ([#60](https://github.com/nf-core/crisprseq/pull/60) +- Change to `process_high` for the mageck mle module ([#60](https://github.com/nf-core/crisprseq/pull/60)) +- Fix paired-end samplesheet file for screening ([#60](https://github.com/nf-core/crisprseq/pull/60)) - Summary processes don't modify the input file anymore, allowing resuming these processes ([#66](https://github.com/nf-core/crisprseq/pull/66)) - Do not stash unexistent files, use empty lists instead. Fixes AWS tests ([#67](https://github.com/nf-core/crisprseq/pull/67)) - Rename process `merging_summary` to `preprocessing_summary` to improve clarity ([#69](https://github.com/nf-core/crisprseq/pull/69)) diff --git a/conf/test_screening.config b/conf/test_screening.config index 0ac8ed89..7661aacf 100644 --- a/conf/test_screening.config +++ b/conf/test_screening.config @@ -25,5 +25,5 @@ params { crisprcleanr = "Brunello_Library" mle_design_matrix = "https://raw.githubusercontent.com/nf-core/test-datasets/crisprseq/testdata/design_matrix.txt" library = "https://raw.githubusercontent.com/nf-core/test-datasets/crisprseq/testdata/brunello_target_sequence.txt" - rra_contrasts = "https://raw.githubusercontent.com/nf-core/test-datasets/crisprseq/testdata/rra_contrasts.txt" + rra_contrasts = "https://raw.githubusercontent.com/nf-core/test-datasets/crisprseq/testdata/rra_contrasts.txt" } diff --git a/nextflow.config b/nextflow.config index 1be05b5b..5f898729 100644 --- a/nextflow.config +++ b/nextflow.config @@ -266,7 +266,7 @@ manifest { mainScript = 'main.nf' nextflowVersion = '!>=23.04.0' version = '2.1.0' - doi = '' + doi = 'https://doi.org/10.5281/zenodo.7598496' } // Load modules.config for DSL2 module specific options From 3818dd28c3e333fd589c641faf9b28c33aff16cc Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Fri, 10 Nov 2023 14:02:29 +0100 Subject: [PATCH 118/133] Add versions for graph rra --- modules/local/mageck/graphrra.nf | 2 +- workflows/crisprseq_screening.nf | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/mageck/graphrra.nf b/modules/local/mageck/graphrra.nf index ec11bf55..4e349b4c 100644 --- a/modules/local/mageck/graphrra.nf +++ b/modules/local/mageck/graphrra.nf @@ -12,7 +12,7 @@ process MAGECK_GRAPHRRA { output: tuple val(meta), path("*.png"), emit: graphs - + path "versions.yml" , emit: versions when: task.ext.when == null || task.ext.when diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index d198f831..6e393238 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -170,7 +170,7 @@ workflow CRISPRSEQ_SCREENING { MAGECK_GRAPHRRA ( MAGECK_TEST.out.gene_summary ) - //TODO: Missing versions + ch_versions = ch_versions.mix(MAGECK_GRAPHRRA.out.versions) } if(params.rra_contrasts) { From f3c6ace64a16399d0c82c44e0448cec9c4089105 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Mon, 13 Nov 2023 09:34:53 +0100 Subject: [PATCH 119/133] Try to add versions --- modules/local/bagel2/graph.nf | 1 - modules/local/bagel2/pr.nf | 2 +- modules/local/clustering_summary.nf | 2 ++ modules/local/preprocessing_summary.nf | 2 ++ modules/nf-core/mageck/count/main.nf | 9 ++++----- templates/clustering_summary.py | 3 +++ templates/preprocessing_summary.py | 8 ++++++++ workflows/crisprseq_screening.nf | 4 +++- workflows/crisprseq_targeted.nf | 4 +++- 9 files changed, 26 insertions(+), 9 deletions(-) diff --git a/modules/local/bagel2/graph.nf b/modules/local/bagel2/graph.nf index 270c265f..62cb12d0 100644 --- a/modules/local/bagel2/graph.nf +++ b/modules/local/bagel2/graph.nf @@ -20,7 +20,6 @@ process BAGEL2_GRAPH { script: def args = task.ext.args ?: '' - def prefix = task.ext.prefix ?: "${meta.id}" """ #!/usr/bin/env python3 diff --git a/modules/local/bagel2/pr.nf b/modules/local/bagel2/pr.nf index cc4f3157..e27366d6 100644 --- a/modules/local/bagel2/pr.nf +++ b/modules/local/bagel2/pr.nf @@ -23,7 +23,7 @@ process BAGEL2_PR { def prefix = task.ext.prefix ?: "${meta.id}" """ - BAGEL.py pr -i $bf -o '${meta.treatment}_vs_${meta.reference}.pr' -e $reference_essentials -n $reference_nonessentials + BAGEL.py pr -i $bf -o '${meta.treatment}_vs_${meta.reference}.pr' -e $reference_essentials -n $reference_nonessentials $args cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/local/clustering_summary.nf b/modules/local/clustering_summary.nf index b31d4f36..a48fa270 100644 --- a/modules/local/clustering_summary.nf +++ b/modules/local/clustering_summary.nf @@ -12,6 +12,8 @@ process CLUSTERING_SUMMARY { output: tuple val(meta), path("*_clustering_summary.csv"), emit: summary + path "versions.yml", emit: versions + when: task.ext.when == null || task.ext.when diff --git a/modules/local/preprocessing_summary.nf b/modules/local/preprocessing_summary.nf index 52d29e17..f1aafa7c 100644 --- a/modules/local/preprocessing_summary.nf +++ b/modules/local/preprocessing_summary.nf @@ -13,6 +13,8 @@ process PREPROCESSING_SUMMARY { output: tuple val(meta), path("*_preprocessing_summary.csv"), emit: summary + path "versions.yml", emit: versions + when: task.ext.when == null || task.ext.when diff --git a/modules/nf-core/mageck/count/main.nf b/modules/nf-core/mageck/count/main.nf index ced51e85..f47ad55e 100644 --- a/modules/nf-core/mageck/count/main.nf +++ b/modules/nf-core/mageck/count/main.nf @@ -12,13 +12,12 @@ process MAGECK_COUNT { path(library) output: - tuple val(meta), path("*count.txt"), emit: count + tuple val(meta), path("*count.txt"), emit: count tuple val(meta), path("*.count_normalized.txt"), emit: norm - tuple val(meta), path("*.countsummary.txt"), emit: summary + tuple val(meta), path("*.countsummary.txt"), emit: summary tuple val(meta), path("*.count_normalized.txt"), emit: normalized - tuple val(meta), path("*.log"), emit: logs - - path "versions.yml" , emit: versions + tuple val(meta), path("*.log"), emit: logs + path "versions.yml", emit: versions when: task.ext.when == null || task.ext.when diff --git a/templates/clustering_summary.py b/templates/clustering_summary.py index 5b1dfd42..33feb1b1 100644 --- a/templates/clustering_summary.py +++ b/templates/clustering_summary.py @@ -22,3 +22,6 @@ add_line = False if add_line: output_file.write(f"clustered-reads, {clusters_count}\\n") + +with open("versions.yml", "w") as f: + f.write('"${task.process}":\\n') diff --git a/templates/preprocessing_summary.py b/templates/preprocessing_summary.py index 10cb94b1..c2a30578 100755 --- a/templates/preprocessing_summary.py +++ b/templates/preprocessing_summary.py @@ -49,3 +49,11 @@ output_file.write( f"quality-filtered-reads, {trimmed_reads_count} ({round(trimmed_reads_count * 100 / assembled_reads_count,1)}%)\\n" ) + + # Output version information + #version = pd.__version__ + #matplotlib_version = plt.matplotlib.__version__ + # alas, no `pyyaml` pre-installed in the cellranger container + with open("versions.yml", "w") as f: + f.write('"${task.process}":\\n') + diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 6e393238..031fe86a 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -128,7 +128,6 @@ workflow CRISPRSEQ_SCREENING { ch_versions = ch_versions.mix(MAGECK_COUNT.out.versions.first()) - MAGECK_COUNT.out.count.map { it -> it[1] }.set { ch_counts } @@ -209,6 +208,9 @@ workflow CRISPRSEQ_SCREENING { BAGEL2_GRAPH ( BAGEL2_PR.out.pr ) + + ch_versions = ch_versions.mix(BAGEL2_GRAPH.out.versions) + } if(params.mle_design_matrix) { diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index a94f939a..87c4cb89 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -601,6 +601,9 @@ workflow CRISPRSEQ_TARGETED { .join(PREPROCESSING_SUMMARY.out.summary) ) + ch_versions = ch_versions.mix(CLUSTERING_SUMMARY.out.versions) + + // // MODULE: Mapping with Minimap2 @@ -702,7 +705,6 @@ workflow CRISPRSEQ_TARGETED { } } .set { ch_template_bam } - ch_versions = ch_versions.mix(MINIMAP2_ALIGN_TEMPLATE.out.versions) ch_mapped_bam .join(SAMTOOLS_INDEX.out.bai) From 1abc804a7e7973231a857ad1b1eed95a5d8ffae7 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Mon, 13 Nov 2023 10:30:49 +0100 Subject: [PATCH 120/133] Update modules/local/bagel2/graph.nf --- modules/local/bagel2/graph.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/modules/local/bagel2/graph.nf b/modules/local/bagel2/graph.nf index 62cb12d0..270c265f 100644 --- a/modules/local/bagel2/graph.nf +++ b/modules/local/bagel2/graph.nf @@ -20,6 +20,7 @@ process BAGEL2_GRAPH { script: def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${meta.id}" """ #!/usr/bin/env python3 From c8ad5a24611b46703739db99c843f57e46201587 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Mon, 13 Nov 2023 13:12:14 +0100 Subject: [PATCH 121/133] Fix typo --- modules/local/mageck/graphrra.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/mageck/graphrra.nf b/modules/local/mageck/graphrra.nf index 4e349b4c..d6d35b66 100644 --- a/modules/local/mageck/graphrra.nf +++ b/modules/local/mageck/graphrra.nf @@ -50,7 +50,7 @@ process MAGECK_GRAPHRRA { version_file_path <- "versions.yml" version_flute <- paste(unlist(packageVersion("MAGeCKFlute")), collapse = ".") - version_ggplot <- paste(unlist(packageVersion("MAGeCKFlute")), collapse = ".") + version_ggplot <- paste(unlist(packageVersion("ggplot2")), collapse = ".") f <- file(version_file_path, "w") writeLines('"${task.process}":', f) From 0034a4d9cb171e7c6d1fa37cece24420ce890bcf Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 13 Nov 2023 13:03:48 +0100 Subject: [PATCH 122/133] add versions to template scripts --- templates/alignment_summary.py | 4 ++++ templates/clustering_summary.py | 5 +++-- templates/preprocessing_summary.py | 17 +++++++---------- 3 files changed, 14 insertions(+), 12 deletions(-) diff --git a/templates/alignment_summary.py b/templates/alignment_summary.py index 12dd6109..a6b12c06 100644 --- a/templates/alignment_summary.py +++ b/templates/alignment_summary.py @@ -22,3 +22,7 @@ add_line = False if add_line: output_file.write(f"aligned-reads, {mapped_reads_count} ({round(mapped_reads_percentage, 1)}%)\\n") + +with open("versions.yml", "w") as f: + f.write('"${task.process}":\\n') + f.write(f' pysam: "{pysam.__version__}"\\n') diff --git a/templates/clustering_summary.py b/templates/clustering_summary.py index 33feb1b1..4981351a 100644 --- a/templates/clustering_summary.py +++ b/templates/clustering_summary.py @@ -3,10 +3,10 @@ import gzip import sys -from Bio import SeqIO +import Bio with gzip.open("$reads", "rt") as handle: - clusters_count = len(list(SeqIO.parse(handle, "fastq"))) + clusters_count = len(list(Bio.SeqIO.parse(handle, "fastq"))) with open("$summary", "r") as summary: summary_lines = summary.readlines() @@ -25,3 +25,4 @@ with open("versions.yml", "w") as f: f.write('"${task.process}":\\n') + f.write(f' biopython: "{Bio.__version__}"\\n') diff --git a/templates/preprocessing_summary.py b/templates/preprocessing_summary.py index c2a30578..2089eb7c 100755 --- a/templates/preprocessing_summary.py +++ b/templates/preprocessing_summary.py @@ -2,19 +2,19 @@ import gzip -from Bio import SeqIO +import Bio with gzip.open("${raw_reads[0]}", "rt") as handle: - raw_reads_count = len(list(SeqIO.parse(handle, "fastq"))) + raw_reads_count = len(list(Bio.SeqIO.parse(handle, "fastq"))) if "$assembled_reads" == "": assembled_reads_count = 0 else: with gzip.open("$assembled_reads", "rt") as handle: - assembled_reads_count = len(list(SeqIO.parse(handle, "fastq"))) # Merged reads R1+R2 + assembled_reads_count = len(list(Bio.SeqIO.parse(handle, "fastq"))) # Merged reads R1+R2 with gzip.open("$trimmed_reads", "rt") as handle: - trimmed_reads_count = len(list(SeqIO.parse(handle, "fastq"))) # Filtered reads + trimmed_reads_count = len(list(Bio.SeqIO.parse(handle, "fastq"))) # Filtered reads if "$trimmed_adapters" == "": adapters_count = 0 @@ -50,10 +50,7 @@ f"quality-filtered-reads, {trimmed_reads_count} ({round(trimmed_reads_count * 100 / assembled_reads_count,1)}%)\\n" ) - # Output version information - #version = pd.__version__ - #matplotlib_version = plt.matplotlib.__version__ - # alas, no `pyyaml` pre-installed in the cellranger container - with open("versions.yml", "w") as f: - f.write('"${task.process}":\\n') +with open("versions.yml", "w") as f: + f.write('"${task.process}":\\n') + f.write(f' biopython: "{Bio.__version__}"\\n') From 2e58467bbf21f56556d1048f65c6107bd90047e8 Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 13 Nov 2023 13:05:59 +0100 Subject: [PATCH 123/133] collect versions --- workflows/crisprseq_targeted.nf | 3 +++ 1 file changed, 3 insertions(+) diff --git a/workflows/crisprseq_targeted.nf b/workflows/crisprseq_targeted.nf index 87c4cb89..abf67bb2 100644 --- a/workflows/crisprseq_targeted.nf +++ b/workflows/crisprseq_targeted.nf @@ -370,6 +370,7 @@ workflow CRISPRSEQ_TARGETED { PREPROCESSING_SUMMARY { ch_preprocessing_summary_data } + ch_versions = ch_versions.mix(PREPROCESSING_SUMMARY.out.versions) if (params.umi_clustering) { @@ -663,6 +664,7 @@ workflow CRISPRSEQ_TARGETED { ch_mapped_bam .join(CLUSTERING_SUMMARY.out.summary) ) + ch_versions = ch_versions.mix(ALIGNMENT_SUMMARY.out.versions) // @@ -705,6 +707,7 @@ workflow CRISPRSEQ_TARGETED { } } .set { ch_template_bam } + ch_versions = ch_versions.mix(MINIMAP2_ALIGN_TEMPLATE.out.versions) ch_mapped_bam .join(SAMTOOLS_INDEX.out.bai) From 69d20c1b74dbc6e567da64ae03870f41ea7e2aab Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 13 Nov 2023 13:21:16 +0100 Subject: [PATCH 124/133] add versions to R scripts --- modules/local/cigar_parser.nf | 8 +++++++- modules/local/crisprseq_plotter.nf | 14 +++++++++++++- 2 files changed, 20 insertions(+), 2 deletions(-) diff --git a/modules/local/cigar_parser.nf b/modules/local/cigar_parser.nf index 5b87bba4..45dfd7d5 100644 --- a/modules/local/cigar_parser.nf +++ b/modules/local/cigar_parser.nf @@ -41,7 +41,13 @@ process CIGAR_PARSER { cat <<-END_VERSIONS > versions.yml "${task.process}": - Rscript: \$(Rscript --version) + seqinr: \$(Rscript -e "cat(paste(packageVersion('seqinr'), collapse='.'))") + Rsamtools: \$(Rscript -e "cat(paste(packageVersion('Rsamtools'), collapse='.'))") + dplyr: \$(Rscript -e "cat(paste(packageVersion('dplyr'), collapse='.'))") + ShortRead: \$(Rscript -e "cat(paste(packageVersion('ShortRead'), collapse='.'))") + jsonlite: \$(Rscript -e "cat(paste(packageVersion('jsonlite'), collapse='.'))") + stringr: \$(Rscript -e "cat(paste(packageVersion('stringr'), collapse='.'))") + plotly: \$(Rscript -e "cat(paste(packageVersion('plotly'), collapse='.'))") END_VERSIONS """ } diff --git a/modules/local/crisprseq_plotter.nf b/modules/local/crisprseq_plotter.nf index 5612260e..c3b0ed62 100644 --- a/modules/local/crisprseq_plotter.nf +++ b/modules/local/crisprseq_plotter.nf @@ -35,7 +35,19 @@ process CRISPRSEQ_PLOTTER { cat <<-END_VERSIONS > versions.yml "${task.process}": - Rscript: \$(Rscript --version) + ggplot2: \$(Rscript -e "cat(paste(packageVersion('ggplot2'), collapse='.'))") + ShortRead: \$(Rscript -e "cat(paste(packageVersion('ShortRead'), collapse='.'))") + plyr: \$(Rscript -e "cat(paste(packageVersion('plyr'), collapse='.'))") + dplyr: \$(Rscript -e "cat(paste(packageVersion('dplyr'), collapse='.'))") + seqinr: \$(Rscript -e "cat(paste(packageVersion('seqinr'), collapse='.'))") + ggpubr: \$(Rscript -e "cat(paste(packageVersion('ggpubr'), collapse='.'))") + ggmsa: \$(Rscript -e "cat(paste(packageVersion('ggmsa'), collapse='.'))") + seqmagick: \$(Rscript -e "cat(paste(packageVersion('seqmagick'), collapse='.'))") + stringr: \$(Rscript -e "cat(paste(packageVersion('stringr'), collapse='.'))") + tidyr: \$(Rscript -e "cat(paste(packageVersion('tidyr'), collapse='.'))") + ggseqlogo: \$(Rscript -e "cat(paste(packageVersion('ggseqlogo'), collapse='.'))") + plotly: \$(Rscript -e "cat(paste(packageVersion('plotly'), collapse='.'))") + cowplot: \$(Rscript -e "cat(paste(packageVersion('cowplot'), collapse='.'))") END_VERSIONS """ } From b134f12d61b0928565d68d90bbd868200723765a Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 13 Nov 2023 13:35:30 +0100 Subject: [PATCH 125/133] add versions channel to alignment_summary --- modules/local/alignment_summary.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/modules/local/alignment_summary.nf b/modules/local/alignment_summary.nf index 3520689f..1947812c 100644 --- a/modules/local/alignment_summary.nf +++ b/modules/local/alignment_summary.nf @@ -12,6 +12,7 @@ process ALIGNMENT_SUMMARY { output: tuple val(meta), path("*_alignment_summary.csv"), emit: summary + path "versions.yml", emit: versions when: task.ext.when == null || task.ext.when From ea081e0c557e019e63ded3ed2efc2a310d40533c Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Mon, 13 Nov 2023 13:48:55 +0100 Subject: [PATCH 126/133] fix import SeqIO --- templates/clustering_summary.py | 3 ++- templates/preprocessing_summary.py | 7 ++++--- 2 files changed, 6 insertions(+), 4 deletions(-) diff --git a/templates/clustering_summary.py b/templates/clustering_summary.py index 4981351a..1f2d7eb8 100644 --- a/templates/clustering_summary.py +++ b/templates/clustering_summary.py @@ -4,9 +4,10 @@ import sys import Bio +from Bio import SeqIO with gzip.open("$reads", "rt") as handle: - clusters_count = len(list(Bio.SeqIO.parse(handle, "fastq"))) + clusters_count = len(list(SeqIO.parse(handle, "fastq"))) with open("$summary", "r") as summary: summary_lines = summary.readlines() diff --git a/templates/preprocessing_summary.py b/templates/preprocessing_summary.py index 2089eb7c..16f8c74a 100755 --- a/templates/preprocessing_summary.py +++ b/templates/preprocessing_summary.py @@ -3,18 +3,19 @@ import gzip import Bio +from Bio import SeqIO with gzip.open("${raw_reads[0]}", "rt") as handle: - raw_reads_count = len(list(Bio.SeqIO.parse(handle, "fastq"))) + raw_reads_count = len(list(SeqIO.parse(handle, "fastq"))) if "$assembled_reads" == "": assembled_reads_count = 0 else: with gzip.open("$assembled_reads", "rt") as handle: - assembled_reads_count = len(list(Bio.SeqIO.parse(handle, "fastq"))) # Merged reads R1+R2 + assembled_reads_count = len(list(SeqIO.parse(handle, "fastq"))) # Merged reads R1+R2 with gzip.open("$trimmed_reads", "rt") as handle: - trimmed_reads_count = len(list(Bio.SeqIO.parse(handle, "fastq"))) # Filtered reads + trimmed_reads_count = len(list(SeqIO.parse(handle, "fastq"))) # Filtered reads if "$trimmed_adapters" == "": adapters_count = 0 From d7fa6291e64d53c83207136e62f1933e84b9c122 Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Mon, 13 Nov 2023 16:04:16 +0100 Subject: [PATCH 127/133] Last todo statement --- docs/usage/screening.md | 3 +-- modules/local/bagel2/graph.nf | 3 --- 2 files changed, 1 insertion(+), 5 deletions(-) diff --git a/docs/usage/screening.md b/docs/usage/screening.md index bdbdd4c9..63fba93e 100644 --- a/docs/usage/screening.md +++ b/docs/usage/screening.md @@ -59,8 +59,7 @@ MAGeCK RRA performs robust ranking aggregation to identify genes that are consis | ----------------------- | ------------------- | | treatment1 | control1 | -A full example can be found here : -#TODO: add link to rawcontent of nf-core test +A full example can be found [here](https://raw.githubusercontent.com/nf-core/test-datasets/crisprseq/testdata/full_test/samplesheet_full.csv). ### MAGeCK mle diff --git a/modules/local/bagel2/graph.nf b/modules/local/bagel2/graph.nf index 270c265f..32c96046 100644 --- a/modules/local/bagel2/graph.nf +++ b/modules/local/bagel2/graph.nf @@ -7,7 +7,6 @@ process BAGEL2_GRAPH { 'https://depot.galaxyproject.org/singularity/mulled-v2-54e0353146eca1531516863e8235bf7385d76663:c9ff1a9eec871c54cbea815eae778da702623978-0': 'biocontainers/mulled-v2-54e0353146eca1531516863e8235bf7385d76663:c9ff1a9eec871c54cbea815eae778da702623978-0' }" - input: tuple val(meta), path(pr) @@ -30,7 +29,6 @@ process BAGEL2_GRAPH { #### #### Orient a reference sequence according to reads orientation. - import pandas as pd import matplotlib.pyplot as plt @@ -69,7 +67,6 @@ process BAGEL2_GRAPH { f.write(f' pandas: "{version}"\\n') f.write(f' matplotlib.pyplot: "{matplotlib_version}"\\n') - """ From 510c563f043ebccf153b3e3939262590b697cc4d Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C3=BAlia=20Mir=20Pedrol?= Date: Mon, 13 Nov 2023 16:08:38 +0100 Subject: [PATCH 128/133] Update docs/usage/screening.md Co-authored-by: LaurenceKuhl --- docs/usage/screening.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/docs/usage/screening.md b/docs/usage/screening.md index bdbdd4c9..17717cf8 100644 --- a/docs/usage/screening.md +++ b/docs/usage/screening.md @@ -59,8 +59,8 @@ MAGeCK RRA performs robust ranking aggregation to identify genes that are consis | ----------------------- | ------------------- | | treatment1 | control1 | -A full example can be found here : -#TODO: add link to rawcontent of nf-core test +A full example can be found [here](https://raw.githubusercontent.com/nf-core/test-datasets/crisprseq/testdata/full_test/samplesheet_full.csv). + ### MAGeCK mle From 8b344a3114ab3da444eecd2603665d25b5794914 Mon Sep 17 00:00:00 2001 From: nf-core-bot Date: Mon, 13 Nov 2023 15:09:50 +0000 Subject: [PATCH 129/133] [automated] Fix linting with Prettier --- docs/usage/screening.md | 1 - 1 file changed, 1 deletion(-) diff --git a/docs/usage/screening.md b/docs/usage/screening.md index 17717cf8..63fba93e 100644 --- a/docs/usage/screening.md +++ b/docs/usage/screening.md @@ -61,7 +61,6 @@ MAGeCK RRA performs robust ranking aggregation to identify genes that are consis A full example can be found [here](https://raw.githubusercontent.com/nf-core/test-datasets/crisprseq/testdata/full_test/samplesheet_full.csv). - ### MAGeCK mle MAGeCK MLE uses a maximum likelihood estimation approach to estimate the effects of gene knockout on cell fitness. It models the read count data of guide RNAs targeting each gene and estimates the dropout probability for each gene. MAGeCK mle requires a design matrix. The design matrix is a `txt` file indicating the effects of different conditions on different samples. From 1c9a8300ac32edc0830a865771f168659b2a5fee Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C3=BAlia=20Mir=20Pedrol?= Date: Tue, 14 Nov 2023 08:32:32 +0100 Subject: [PATCH 130/133] Apply suggestions from code review Co-authored-by: Friederike Hanssen --- README.md | 3 ++- modules/local/bagel2/graph.nf | 1 - modules/local/mageck/graphrra.nf | 4 ---- 3 files changed, 2 insertions(+), 6 deletions(-) diff --git a/README.md b/README.md index ffc72899..b9f2281e 100644 --- a/README.md +++ b/README.md @@ -1,6 +1,7 @@ # ![nf-core/crisprseq](docs/images/nf-core-crisprseq_logo_light.png#gh-light-mode-only) ![nf-core/crisprseq](docs/images/nf-core-crisprseq_logo_dark.png#gh-dark-mode-only) -[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.7598496-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.7598496) +[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results) +[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.7598496-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.7598496) [![GitHub Actions CI Status](https://github.com/nf-core/crisprseq/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/crisprseq/actions?query=workflow%3A%22nf-core+CI%22) [![GitHub Actions Linting Status](https://github.com/nf-core/crisprseq/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/crisprseq/actions?query=workflow%3A%22nf-core+linting%22)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/crisprseq/results) diff --git a/modules/local/bagel2/graph.nf b/modules/local/bagel2/graph.nf index 270c265f..355d2f43 100644 --- a/modules/local/bagel2/graph.nf +++ b/modules/local/bagel2/graph.nf @@ -7,7 +7,6 @@ process BAGEL2_GRAPH { 'https://depot.galaxyproject.org/singularity/mulled-v2-54e0353146eca1531516863e8235bf7385d76663:c9ff1a9eec871c54cbea815eae778da702623978-0': 'biocontainers/mulled-v2-54e0353146eca1531516863e8235bf7385d76663:c9ff1a9eec871c54cbea815eae778da702623978-0' }" - input: tuple val(meta), path(pr) diff --git a/modules/local/mageck/graphrra.nf b/modules/local/mageck/graphrra.nf index d6d35b66..69f48ae7 100644 --- a/modules/local/mageck/graphrra.nf +++ b/modules/local/mageck/graphrra.nf @@ -59,10 +59,6 @@ process MAGECK_GRAPHRRA { writeLines(" ggplot2: ", f, sep = "") writeLines(version_ggplot, f) close(f) - - - - """ From 23caebbab5e97b8144215907f9433541ebda9ebc Mon Sep 17 00:00:00 2001 From: mirpedrol Date: Tue, 14 Nov 2023 08:34:11 +0100 Subject: [PATCH 131/133] add licenses to template scripts --- templates/alignment_summary.py | 6 ++++++ templates/clustering_summary.py | 6 ++++++ templates/preprocessing_summary.py | 6 ++++++ 3 files changed, 18 insertions(+) diff --git a/templates/alignment_summary.py b/templates/alignment_summary.py index a6b12c06..035e7b90 100644 --- a/templates/alignment_summary.py +++ b/templates/alignment_summary.py @@ -1,5 +1,11 @@ #!/usr/bin/env python +############################ +#### Summary of alignment +#### author: Júlia Mir @mirpedrol +#### Released under the MIT license. See git repository (https://github.com/nf-core/crisprseq) for full license text. +############################ + import sys import pysam diff --git a/templates/clustering_summary.py b/templates/clustering_summary.py index 1f2d7eb8..b1ad29ed 100644 --- a/templates/clustering_summary.py +++ b/templates/clustering_summary.py @@ -1,5 +1,11 @@ #!/usr/bin/env python +############################ +#### Summary of clustering +#### author: Júlia Mir @mirpedrol +#### Released under the MIT license. See git repository (https://github.com/nf-core/crisprseq) for full license text. +############################ + import gzip import sys diff --git a/templates/preprocessing_summary.py b/templates/preprocessing_summary.py index 16f8c74a..13c698ce 100755 --- a/templates/preprocessing_summary.py +++ b/templates/preprocessing_summary.py @@ -1,5 +1,11 @@ #!/usr/bin/env python +############################ +#### Summary of reads preprocessing +#### author: Júlia Mir @mirpedrol +#### Released under the MIT license. See git repository (https://github.com/nf-core/crisprseq) for full license text. +############################ + import gzip import Bio From fd384fa07623aa34c1753aef6c09b6b1a1aaa81e Mon Sep 17 00:00:00 2001 From: LaurenceKuhl Date: Tue, 14 Nov 2023 11:17:21 +0100 Subject: [PATCH 132/133] Fix version collection --- workflows/crisprseq_screening.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/crisprseq_screening.nf b/workflows/crisprseq_screening.nf index 031fe86a..e37619ce 100644 --- a/workflows/crisprseq_screening.nf +++ b/workflows/crisprseq_screening.nf @@ -229,7 +229,7 @@ workflow CRISPRSEQ_SCREENING { } CUSTOM_DUMPSOFTWAREVERSIONS ( - ch_versions.unique().collectFile(name: 'collated_versions.yml') + ch_versions.unique{ it.text }.collectFile(name: 'collated_versions.yml') ) // From 659177368b3b1bbe049a8d896f541d48ca588aca Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C3=BAlia=20Mir=20Pedrol?= Date: Tue, 14 Nov 2023 12:31:34 +0100 Subject: [PATCH 133/133] Apply suggestions from code review --- CHANGELOG.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 9f846e3b..0b8f63bc 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ 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). -## [v2.1.0 - Jamon Salas](https://github.com/nf-core/crisprseq/releases/tag/2.1.0) - [08.11.2023] +## [v2.1.0 - Jamon Salas](https://github.com/nf-core/crisprseq/releases/tag/2.1.0) - [14.11.2023] ### Added @@ -11,6 +11,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - Use `Channel.fromSamplesheet()` from `nf-validation` to validate input sample sheets and create an input channel ([#58](https://github.com/nf-core/crisprseq/pull/58)) - BAGEL2 as a module which detects gene essentiality ([#60](https://github.com/nf-core/crisprseq/pull/60)) - Add custom plots to MultiQC report (cutadapt module, read processing, edition, edition QC) ([#64](https://github.com/nf-core/crisprseq/pull/64)) +- Template update v2.10 ([#79](https://github.com/nf-core/crisprseq/pull/79)) ### Fixed