From e1b45110c6faf890be1f00d745f092f615d5cf1f Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Thu, 28 Sep 2023 14:27:02 -0400 Subject: [PATCH 001/100] chore: create module & subworkflow scaffolding for prepare_genome --- modules/local/prepare_genome.nf | 0 subworkflows/local/prepare_genome.nf | 4 ++++ 2 files changed, 4 insertions(+) create mode 100644 modules/local/prepare_genome.nf create mode 100644 subworkflows/local/prepare_genome.nf diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf new file mode 100644 index 00000000..e69de29b diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf new file mode 100644 index 00000000..b465acaa --- /dev/null +++ b/subworkflows/local/prepare_genome.nf @@ -0,0 +1,4 @@ +workflow PREPARE_GENOME { + ch_fasta = file(params.fasta) + ch_gtf = file(params.fasta) +} From 691349ca799477ea338ea22fa3203eb515b84e91 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 09:07:05 -0400 Subject: [PATCH 002/100] feat: support paired-end reads in ALIGN_BLACKLIST towards #24 --- modules/local/align.nf | 13 +++++++------ 1 file changed, 7 insertions(+), 6 deletions(-) diff --git a/modules/local/align.nf b/modules/local/align.nf index 69598378..c91bb9b8 100644 --- a/modules/local/align.nf +++ b/modules/local/align.nf @@ -13,17 +13,18 @@ process ALIGN_BLACKLIST { output: tuple val(meta), path("${meta.id}.no_blacklist.fastq.gz"), emit: reads - script: // TODO use samtools -f4 for single-end and -f12 for paired to get unmapped reads https://broadinstitute.github.io/picard/explain-flags.html + script: def prefix = task.ext.prefix ?: "${meta.id}" + def filter_flag = meta.single_end ? '4' : '12' """ bwa mem -t $task.cpus ${params.genomes[ params.genome ].blacklist} $fastq > ${prefix}.sam samtools view \\ - -@ ${task.cpus} \\ - -f4 \\ - -b \\ - ${prefix}.sam |\ + -@ ${task.cpus} \\ + -f${filter_flag} \\ + -b \\ + ${prefix}.sam |\ samtools bam2fq |\ - pigz -p $task.cpus > ${prefix}.no_blacklist.fastq.gz + pigz -p $task.cpus > ${prefix}.no_blacklist.fastq.gz """ stub: From c218d806b78d73e86312047c5707dad3b55fcb30 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 09:15:55 -0400 Subject: [PATCH 003/100] feat: use tmpdir in ppqt --- modules/local/qc.nf | 7 ++++++- 1 file changed, 6 insertions(+), 1 deletion(-) diff --git a/modules/local/qc.nf b/modules/local/qc.nf index 0bab8a45..19c31196 100644 --- a/modules/local/qc.nf +++ b/modules/local/qc.nf @@ -156,8 +156,13 @@ process PHANTOM_PEAKS { script: // TODO: for PE, just use first read of each pair def prefix = task.ext.prefix ?: "${meta.id}" """ + # current working directory is a tmpdir when 'scratch' is set + TMP=tmp/ + mkdir \$TMP + trap 'rm -rf "\$TMP"' EXIT + RUN_SPP=\$(which run_spp.R) - Rscript \$RUN_SPP -c=${bam} -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out + Rscript \$RUN_SPP -c=${bam} -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP frag_len=`cut -f 3 ${prefix}.spp.out | sed 's/,.*//g'` echo \$frag_len > "${meta.id}.fraglen.txt" """ From 6a79397b9d70075190a2e165cb765bf227719be2 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 09:36:58 -0400 Subject: [PATCH 004/100] feat: support paired end & tmpdir in ppqt --- assets/dag.png | Bin 258554 -> 108630 bytes conf/test.config | 2 +- modules/local/qc.nf | 53 +++++++++++++++++++++++++++++++------------- 3 files changed, 39 insertions(+), 16 deletions(-) diff --git a/assets/dag.png b/assets/dag.png index 8b7b53ee11fc3ba999e5226ba98cf8ae5140923c..f3ef3e94066f247218465b7f49c1d9cc1c754644 100644 GIT binary patch literal 108630 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a/modules/local/qc.nf b/modules/local/qc.nf index 19c31196..3c0c2f86 100644 --- a/modules/local/qc.nf +++ b/modules/local/qc.nf @@ -138,7 +138,6 @@ process PARSE_PRESEQ_LOG { } process PHANTOM_PEAKS { - // TODO: set tmpdir as lscratch if available https://github.com/CCBR/Pipeliner/blob/86c6ccaa3d58381a0ffd696bbf9c047e4f991f9e/Rules/InitialChIPseqQC.snakefile#L504 tag { meta.id } label 'qc' label 'ppqt' @@ -149,23 +148,47 @@ process PHANTOM_PEAKS { tuple val(meta), path(bam), path(bai) output: - path("${meta.id}.ppqt.pdf"), emit: pdf - path("${meta.id}.spp.out"), emit: spp + path("${meta.id}.ppqt.pdf") , emit: pdf + path("${meta.id}.spp.out") , emit: spp tuple val(meta), path("${meta.id}.fraglen.txt"), emit: fraglen + path "versions.yml" , emit: versions - script: // TODO: for PE, just use first read of each pair + script: def prefix = task.ext.prefix ?: "${meta.id}" - """ - # current working directory is a tmpdir when 'scratch' is set - TMP=tmp/ - mkdir \$TMP - trap 'rm -rf "\$TMP"' EXIT - - RUN_SPP=\$(which run_spp.R) - Rscript \$RUN_SPP -c=${bam} -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP - frag_len=`cut -f 3 ${prefix}.spp.out | sed 's/,.*//g'` - echo \$frag_len > "${meta.id}.fraglen.txt" - """ + def VERSION = '1.2.2' // WARN: Version information not provided by tool on CLI. Please update this string when bumping container versions. + def prep_env = """ + # current working directory is a tmpdir when 'scratch' is set + TMP=tmp/ + mkdir \$TMP + trap 'rm -rf "\$TMP"' EXIT + # get spp path + RUN_SPP=\$(which run_spp.R) + """ + def cleanup = """ + # get fragment length + frag_len=`cut -f 3 ${prefix}.spp.out | sed 's/,.*//g'` + echo \$frag_len > "${meta.id}.fraglen.txt" + # export versions + cat <<-END_VERSIONS > versions.yml + "${task.process}": + phantompeakqualtools: $VERSION + END_VERSIONS + """ + if (meta.single_end) { + """ + ${prep_env} + Rscript \$RUN_SPP -c=${bam} -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP + ${cleanup} + """ + } else { // for PE, just use first read of each pair + """ + ${prep_env} + samtools view -b -f 66 -o \$TMP/${bam.baseName}.f66.bam ${bam} + samtools index \$TMP/${bam.baseName}.f66.bam + Rscript \$RUN_SPP -c=\$TMP/${bam.baseName}.f66.bam -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP + ${cleanup} + """ + } stub: """ From 73c6cdc455070e86c4ace33328d8c7fd9b873b33 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 10:58:50 -0400 Subject: [PATCH 005/100] feat: use Picard markduplicates for paired end; refactor with dedup as separate subworkflow --- conf/containers.config | 1 + main.nf | 12 ++-- modules/local/deduplicate.nf | 91 +++++++++++++++++++++++++++++++ modules/local/qc.nf | 52 ------------------ modules/local/samtools_index.nf | 36 ++++++++++++ subworkflows/local/deduplicate.nf | 35 ++++++++++++ 6 files changed, 169 insertions(+), 58 deletions(-) create mode 100644 modules/local/deduplicate.nf create mode 100644 modules/local/samtools_index.nf create mode 100644 subworkflows/local/deduplicate.nf diff --git a/conf/containers.config b/conf/containers.config index 8387c517..b334600d 100644 --- a/conf/containers.config +++ b/conf/containers.config @@ -11,6 +11,7 @@ params { multiqc = 'nciccbr/ccbr_multiqc_1.15:v1' ngsqc = 'nciccbr/ccbr_ngsqc_0.31:v1' phantom_peaks = 'quay.io/biocontainers/phantompeakqualtools:1.2.2--hdfd78af_1' + picard = 'quay.io/biocontainers/picard:2.27.4--hdfd78af_0' preseq = 'nciccbr/ccbr_preseq_v2.0:v1' r = 'nciccbr/ccbr_r_4.3.0:v1' sicer = 'nciccbr/ccbr_sicer2_1.0.3:v1' diff --git a/main.nf b/main.nf index 34c14339..e03dc482 100644 --- a/main.nf +++ b/main.nf @@ -18,15 +18,15 @@ log.info """\ .stripIndent() // SUBWORKFLOWS -include { INPUT_CHECK } from './subworkflows/local/input_check.nf' -include { QC } from './subworkflows/local/qc.nf' -include { CALL_PEAKS } from './subworkflows/local/peaks.nf' +include { INPUT_CHECK } from './subworkflows/local/input_check.nf' +include { DEDUPLICATE } from "./subworkflows/local/deduplicate.nf" +include { QC } from './subworkflows/local/qc.nf' +include { CALL_PEAKS } from './subworkflows/local/peaks.nf' // MODULES -include { CUTADAPT } from "./modules/CCBR/cutadapt" +include { CUTADAPT } from "./modules/CCBR/cutadapt" include { ALIGN_BLACKLIST } from "./modules/local/align.nf" include { ALIGN_GENOME } from "./modules/local/align.nf" -include { DEDUPLICATE } from "./modules/local/qc.nf" include { PHANTOM_PEAKS } from "./modules/local/qc.nf" include { PPQT_PROCESS MULTIQC } from "./modules/local/qc.nf" @@ -51,7 +51,7 @@ workflow { Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) .set{ chrom_sizes } - aligned_bam.combine(chrom_sizes) | DEDUPLICATE + DEDUPLICATE(aligned_bam, chrom_sizes) DEDUPLICATE.out.bam.set{ deduped_bam } DEDUPLICATE.out.tag_align.set{ deduped_tagalign } diff --git a/modules/local/deduplicate.nf b/modules/local/deduplicate.nf new file mode 100644 index 00000000..7beb369e --- /dev/null +++ b/modules/local/deduplicate.nf @@ -0,0 +1,91 @@ + +process MACS2_DEDUP { + tag { meta.id } + label 'dedup' + label 'process_medium' + + container = "${params.containers.macs2}" + + input: + tuple val(meta), path(bam), path(chrom_sizes) + + output: + tuple val(meta), path("${meta.id}.TagAlign.dedup.bed"), emit: bed + tuple val(meta), path("${meta.id}.TagAlign.dedup.bam"), emit: bam + + script: + """ + # current working directory is a tmpdir when 'scratch' is set + TMP=tmp + mkdir \$TMP + trap 'rm -rf "\$TMP"' EXIT + + macs2 filterdup -i ${bam} -g ${params.genomes[ params.genome ].effective_genome_size} --keep-dup="auto" -o TmpTagAlign1 + awk -F"\\t" -v OFS="\\t" '{{if (\$2>0 && \$3>0) {{print}}}}' TmpTagAlign1 > TmpTagAlign2 + awk -F"\\t" -v OFS="\\t" '{{print \$1,1,\$2}}' ${chrom_sizes} > \$TMP/GenomeFile_unsorted.bed + sort \\ + -k1,1 -k2,2n \\ + -T \$TMP \\ + -S 2G \\ + --parallel ${task.cpus} \\ + \$TMP/GenomeFile_unsorted.bed > GenomeFile.bed + bedtools intersect -wa -f 1.0 -a TmpTagAlign2 -b GenomeFile.bed | awk -F"\\t" -v OFS="\\t" '{\$5="0"; print}' > ${meta.id}.TagAlign.dedup.bed + bedtools bedtobam -i ${meta.id}.TagAlign.dedup.bed -g ${chrom_sizes} > ${meta.id}.TagAlign.dedup.bam + """ + + stub: + """ + for ext in bed bam; do + touch ${meta.id}.TagAlign.dedup.\$ext + done + """ +} + +process PICARD_DEDUP { + tag { meta.id } + label 'dedup' + label 'process_medium' + + container = "${params.containers.picard}" + + input: + tuple val(meta), path(bam) + + output: + tuple val(meta), path("${bam.baseName}.dedup.bam") , emit: bam + tuple val(meta), path("${bam.baseName}.MarkDuplicates.metrics.txt"), emit: metrics + path "versions.yml" , emit: versions + + script: + """ + # current working directory is a tmpdir when 'scratch' is set + TMP=tmp + mkdir \$TMP + trap 'rm -rf "\$TMP"' EXIT + + picard -Xmx${task.memory.toGiga()}G \\ + MarkDuplicates \\ + --INPUT ${bam} \\ + --OUTPUT ${bam.baseName}.dedup.bam \\ + --TMP_DIR \$TMP \\ + --VALIDATION_STRINGENCY SILENT \\ + --REMOVE_DUPLICATES true \\ + --METRICS_FILE ${bam.baseName}.MarkDuplicates.metrics.txt + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + picard: \$(echo \$(picard MarkDuplicates --version 2>&1) | grep -o 'Version:.*' | cut -f2- -d:) + END_VERSIONS + """ + + stub: + """ + for ext in dedup.bam MarkDuplicates.metrics.txt; do + touch ${bam.baseName}.\$ext + done + cat <<-END_VERSIONS > versions.yml + "${task.process}": + picard: \$(echo \$(picard MarkDuplicates --version 2>&1) | grep -o 'Version:.*' | cut -f2- -d:) + END_VERSIONS + """ +} diff --git a/modules/local/qc.nf b/modules/local/qc.nf index 3c0c2f86..1e5ff261 100644 --- a/modules/local/qc.nf +++ b/modules/local/qc.nf @@ -221,58 +221,6 @@ process PPQT_PROCESS { """ } -process DEDUPLICATE { - tag { meta.id } - label 'qc' - label 'process_medium' - - container = "${params.containers.macs2}" - - input: - tuple val(meta), path(bam), path(chrom_sizes) - - output: - tuple val(meta), path("${meta.id}.TagAlign.bed"), emit: tag_align - tuple val(meta), path("${bam.baseName}.dedup.bam"), path("${bam.baseName}.dedup.bam.bai"), emit: bam - tuple path("${bam.baseName}.dedup.bam.flagstat"), path("${bam.baseName}.dedup.bam.idxstat"), emit: flagstat - - script: - """ - # current working directory is a tmpdir when 'scratch' is set - TMP=tmp - mkdir \$TMP - trap 'rm -rf "\$TMP"' EXIT - - macs2 filterdup -i ${bam} -g ${params.genomes[ params.genome ].effective_genome_size} --keep-dup="auto" -o TmpTagAlign1 - awk -F"\\t" -v OFS="\\t" '{{if (\$2>0 && \$3>0) {{print}}}}' TmpTagAlign1 > TmpTagAlign2 - awk -F"\\t" -v OFS="\\t" '{{print \$1,1,\$2}}' ${chrom_sizes} > \$TMP/GenomeFile_unsorted.bed - sort \\ - -k1,1 -k2,2n \\ - -T \$TMP \\ - -S 2G \\ - --parallel ${task.cpus} \\ - \$TMP/GenomeFile_unsorted.bed > GenomeFile.bed - bedtools intersect -wa -f 1.0 -a TmpTagAlign2 -b GenomeFile.bed | awk -F"\\t" -v OFS="\\t" '{\$5="0"; print}' > ${meta.id}.TagAlign.bed - bedtools bedtobam -i ${meta.id}.TagAlign.bed -g ${chrom_sizes} > \$TMP/${meta.id}.TagAlign.bed.bam - samtools sort \\ - -@ ${task.cpus} \\ - -m 2G \\ - -T \$TMP \\ - \$TMP/${meta.id}.TagAlign.bed.bam > ${bam.baseName}.dedup.bam - samtools index ${bam.baseName}.dedup.bam - samtools flagstat ${bam.baseName}.dedup.bam > ${bam.baseName}.dedup.bam.flagstat - samtools idxstats ${bam.baseName}.dedup.bam > ${bam.baseName}.dedup.bam.idxstat - """ - - stub: - """ - touch ${meta.id}.TagAlign.bed - for ext in dedup.bam dedup.bam.bai dedup.bam.flagstat dedup.bam.idxstat; do - touch ${bam.baseName}.\${ext} - done - """ -} - process NGSQC_GEN { // TODO segfault - https://github.com/CCBR/CHAMPAGNE/issues/13 tag { meta.id } label 'qc' diff --git a/modules/local/samtools_index.nf b/modules/local/samtools_index.nf new file mode 100644 index 00000000..997041d3 --- /dev/null +++ b/modules/local/samtools_index.nf @@ -0,0 +1,36 @@ +process SAMTOOLS_INDEX { + tag { meta.id } + label 'process_medium' + + container = "${params.containers.base}" + + input: + tuple val(meta), path(bam) + + output: + tuple val(meta), path("${bam.baseName}.sort.bam"), path("${bam.baseName}.sort.bam.bai"), emit: bam + tuple path("${bam.baseName}.sort.bam.flagstat"), path("${bam.baseName}.sort.bam.idxstat"), emit: flagstat + + script: + """ + # current working directory is a tmpdir when 'scratch' is set + TMP=tmp + mkdir \$TMP + trap 'rm -rf "\$TMP"' EXIT + + samtools sort \\ + -@ ${task.cpus} \\ + -m 2G \\ + -T \$TMP \\ + ${bam} > ${bam.baseName}.sort.bam + samtools index ${bam.baseName}.sort.bam + samtools flagstat ${bam.baseName}.sort.bam > ${bam.baseName}.sort.bam.flagstat + samtools idxstats ${bam.baseName}.sort.bam > ${bam.baseName}.sort.bam.idxstat + """ + stub: + """ + for ext in sort.bam sort.bam.bai sort.bam.flagstat sort.bam.idxstat; do + touch ${bam.baseName}.\${ext} + done + """ +} diff --git a/subworkflows/local/deduplicate.nf b/subworkflows/local/deduplicate.nf new file mode 100644 index 00000000..9552670b --- /dev/null +++ b/subworkflows/local/deduplicate.nf @@ -0,0 +1,35 @@ +include { MACS2_DEDUP; PICARD_DEDUP } from "../../modules/local/deduplicate.nf" +include { SAMTOOLS_INDEX as INDEX_SINGLE + SAMTOOLS_INDEX as INDEX_PAIRED } from "../../modules/local/samtools_index.nf" +workflow DEDUPLICATE { + take: + aligned_bam + chrom_sizes + + main: + + // branch reads: single end to macs2 filterdup, paired end to picard markduplicates + aligned_bam.branch { meta, bam -> + single: meta.single_end + paired: !meta.single_end + + }.set{ bam } + bam.single.combine(chrom_sizes) | MACS2_DEDUP + MACS2_DEDUP.out.bam | INDEX_SINGLE + bam.paired | PICARD_DEDUP + PICARD_DEDUP.out.bam | INDEX_PAIRED + INDEX_PAIRED.out.bam + .mix(INDEX_SINGLE.out.bam) + .set{ ch_bam_bai } + + // mix single bed and paired bam channel for peak callers, with variable to designate format + INDEX_PAIRED.out.bam.combine(Channel.value("BAMPE")) + .mix(MACS2_DEDUP.out.bed.combine(Channel.value("NO_BAI")).combine(Channel.value("TagAlign"))) + .set{ mixed_tagAlign } + mixed_tagAlign | view + + + emit: + bam = ch_bam_bai + tag_align = mixed_tagAlign +} From 18a4b0f5c10a1e4b94ac7260701ec5211748d82b Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 11:49:00 -0400 Subject: [PATCH 006/100] refactor: likely don't need bai w/ picard bam --- subworkflows/local/deduplicate.nf | 10 +++++++--- 1 file changed, 7 insertions(+), 3 deletions(-) diff --git a/subworkflows/local/deduplicate.nf b/subworkflows/local/deduplicate.nf index 9552670b..188e342a 100644 --- a/subworkflows/local/deduplicate.nf +++ b/subworkflows/local/deduplicate.nf @@ -23,12 +23,16 @@ workflow DEDUPLICATE { .set{ ch_bam_bai } // mix single bed and paired bam channel for peak callers, with variable to designate format - INDEX_PAIRED.out.bam.combine(Channel.value("BAMPE")) - .mix(MACS2_DEDUP.out.bed.combine(Channel.value("NO_BAI")).combine(Channel.value("TagAlign"))) + INDEX_PAIRED.out.bam + .map{ meta, bam, bai -> + [ meta, bam ] + } + .combine(Channel.value("BAMPE")) + .mix(MACS2_DEDUP.out.bed.combine(Channel.value("BED"))) .set{ mixed_tagAlign } + // [meta, bed/bam, bampe/bed] mixed_tagAlign | view - emit: bam = ch_bam_bai tag_align = mixed_tagAlign From 72ffc5b472f6c3b953eaa8af8ce2d92346622c8f Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 12:10:18 -0400 Subject: [PATCH 007/100] feat: use paired test dataset --- conf/test_paired.config | 21 +++++++++++++++++++++ nextflow.config | 3 +++ 2 files changed, 24 insertions(+) diff --git a/conf/test_paired.config b/conf/test_paired.config index 8c4c2dee..5f616304 100644 --- a/conf/test_paired.config +++ b/conf/test_paired.config @@ -4,4 +4,25 @@ params { outdir = 'results/test_paired' input = 'https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/samplesheet/v2.0/samplesheet_test.csv' + genome = 'hg38' + + max_cpus = 32 // for interactive node on biowulf + max_memory = '120 GB' // for interactive node on biowulf + + deeptools.bin_size = 10000 // this value is only to make bamCoverage run faster. use smaller value for real data. + run { + qc = false + normalize_input = false + call_peaks = false // no peaks exist with this yeast dataset with hg38 as the reference + } +} +dag { + enabled = true + overwrite = true + file = "assets/dag.png" +} +report { + enabled = true + overwrite = true + file = "${params.outdir}/pipeline_info/execution_report.html" } diff --git a/nextflow.config b/nextflow.config index 91f50bbe..9aed1821 100644 --- a/nextflow.config +++ b/nextflow.config @@ -87,6 +87,9 @@ profiles { test { includeConfig "conf/test.config" } + test_paired { + includeConfig "conf/test_paired.config" + } test_mm10 { includeConfig "conf/test_mm10.config" } From 305ab38bbb93c6d1bebc9bc8aa88fe2f634d5aaf Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 12:13:06 -0400 Subject: [PATCH 008/100] feat: accept paired end reads for all peak callers --- modules/local/bedtools.nf | 20 ++++++++++++++ modules/local/peaks.nf | 12 +++++---- subworkflows/local/deduplicate.nf | 4 +-- subworkflows/local/peaks.nf | 45 ++++++++++++++++++++++++------- 4 files changed, 63 insertions(+), 18 deletions(-) create mode 100644 modules/local/bedtools.nf diff --git a/modules/local/bedtools.nf b/modules/local/bedtools.nf new file mode 100644 index 00000000..4fe44c7d --- /dev/null +++ b/modules/local/bedtools.nf @@ -0,0 +1,20 @@ +process BAM_TO_BED { + tag { meta.id } + + container "${params.containers.base}" + + input: + tuple val(meta), path(bam) + + output: + tuple val(meta), path(bed), emit: bed + + script: + """ + bedtools bamtobed -i ${bam} > ${bam.baseName}.bed + """ + stub: + """ + touch ${bam.baseName}.bed + """ +} diff --git a/modules/local/peaks.nf b/modules/local/peaks.nf index 4a320424..83584e21 100644 --- a/modules/local/peaks.nf +++ b/modules/local/peaks.nf @@ -31,7 +31,7 @@ process MACS_BROAD { container = "${params.containers.macs2}" input: - tuple val(meta), path(chip), path(input), val(fraglen), val(genome_frac) + tuple val(meta), path(chip), path(input), val(format), val(fraglen), val(genome_frac) output: tuple val(meta), path("${meta.id}_peaks.broadPeak"), val("${task.process.tokenize(':')[-1].toLowerCase()}"), emit: peak @@ -49,7 +49,7 @@ process MACS_BROAD { --nomodel \\ -q ${params.macs.broad.q} \\ --keep-dup='all' \\ - --format BED \\ + --format ${format} \\ --broad \\ --broad-cutoff ${params.macs.broad.cutoff} """ @@ -70,7 +70,7 @@ process MACS_NARROW { container = "${params.containers.macs2}" input: - tuple val(meta), path(chip), path(input), val(fraglen), val(genome_frac) + tuple val(meta), path(chip), path(input), val(format), val(fraglen), val(genome_frac) output: tuple val(meta), path("${meta.id}_peaks.narrowPeak"), val("${task.process.tokenize(':')[-1].toLowerCase()}"), emit: peak @@ -88,7 +88,7 @@ process MACS_NARROW { --nomodel \\ -q ${params.macs.narrow.q} \\ --keep-dup='all' \\ - --format BED + --format ${format} """ stub: @@ -217,7 +217,7 @@ process GEM { container = "${params.containers.gem}" input: - tuple val(meta), path(chip), path(input), path(read_dists), path(chrom_sizes), path(chrom_dir) + tuple val(meta), path(chip), path(input), val(format), path(read_dists), path(chrom_sizes), path(chrom_dir) output: tuple val(meta), path("${meta.id}/*GEM_events.narrowPeak"), val("${task.process.tokenize(':')[-1].toLowerCase()}"), emit: peak @@ -225,6 +225,7 @@ process GEM { script: // $GEMJAR is defined in the docker container + def f = format == 'BAMPE' ? 'SAM' : format """ java -Xmx${task.memory.toGiga()}G -jar \$GEMJAR \\ --t ${task.cpus} \\ @@ -234,6 +235,7 @@ process GEM { --s ${params.genomes[ params.genome ].effective_genome_size} \\ --expt ${chip} \\ --ctrl ${input} \\ + --f ${f} \\ --out ${meta.id} \\ --fold ${params.gem.fold} \\ --k_min ${params.gem.k_min} \\ diff --git a/subworkflows/local/deduplicate.nf b/subworkflows/local/deduplicate.nf index 188e342a..35d660cd 100644 --- a/subworkflows/local/deduplicate.nf +++ b/subworkflows/local/deduplicate.nf @@ -29,9 +29,7 @@ workflow DEDUPLICATE { } .combine(Channel.value("BAMPE")) .mix(MACS2_DEDUP.out.bed.combine(Channel.value("BED"))) - .set{ mixed_tagAlign } - // [meta, bed/bam, bampe/bed] - mixed_tagAlign | view + .set{ mixed_tagAlign } // [meta, bed/bam, bampe/bed] emit: bam = ch_bam_bai diff --git a/subworkflows/local/peaks.nf b/subworkflows/local/peaks.nf index bcec880b..7abec492 100644 --- a/subworkflows/local/peaks.nf +++ b/subworkflows/local/peaks.nf @@ -14,6 +14,7 @@ include { CALC_GENOME_FRAC GET_PEAK_META CONCAT_PEAK_META PLOT_PEAK_WIDTHS } from "../../modules/local/peaks.nf" +include { BAM_TO_BED } from "../../modules/local/bedtools.nf" workflow CALL_PEAKS { @@ -26,31 +27,55 @@ workflow CALL_PEAKS { main: // peak calling genome_frac = CALC_GENOME_FRAC(chrom_sizes) - // create channel with [ meta, chip_tag, input_tag, fraglen, genome_frac] + + // create channel with [ meta, chip_tag, input_tag, format ] deduped_tagalign .combine(deduped_tagalign) .map { - meta1, tag1, meta2, tag2 -> - meta1.control == meta2.id ? [ meta1, tag1, tag2 ]: null + meta1, tag1, format1, meta2, tag2, format2 -> + meta1.control == meta2.id && format1 == format2 ? [ meta1, tag1, tag2, format1 ]: null } + .set{ ch_tagalign } + + // create macs channel with [ meta, chip_tag, input_tag, format, fraglen, genome_frac] + ch_tagalign .join(frag_lengths) .combine(genome_frac) - .set { ch_tagalign } + .set { ch_macs } + // create sicer channel containing only bed files with [ meta, chip_tag, input_tag, fraglen, genome_frac] deduped_tagalign - .combine(deduped_tagalign) + .branch{ meta, tag, format -> + bam: format == 'BAMPE' + return(tuple(meta, tag)) + bed: format == 'BED' + return(tuple(meta, tag)) + }.set{ tag_split } + tag_split.bam | BAM_TO_BED + BAM_TO_BED.out.bed.set{ tag_split_converted } + tag_split.bed.mix(tag_split_converted).set{ tag_all_bed } + tag_all_bed + .combine(tag_all_bed) .map { - meta1, tag1, meta2, tag2 -> - meta1.control == meta2.id ? [ meta1, tag1, tag2 ]: null + meta1, tag1, format1, meta2, tag2, format2 -> + meta1.control == meta2.id && format1 == format2 ? [ meta1, tag1, tag2 ]: null } + .join(frag_lengths) + .combine(genome_frac) + .set { ch_sicer } + + + // create gem channel with [ meta, chip_tag, input_tag, format, read_dists, chrom_sizes, chrom_dir ] + ch_tagalign .combine(Channel.fromPath(params.gem.read_dists, checkIfExists: true)) .combine(chrom_sizes) .combine(Channel.fromPath("${params.genomes[ params.genome ].chromosomes_dir}", type: 'dir', checkIfExists: true)) .set { ch_gem } - ch_tagalign | MACS_BROAD - ch_tagalign | MACS_NARROW - ch_tagalign | SICER | CONVERT_SICER + + ch_macs | MACS_BROAD + ch_macs | MACS_NARROW + ch_sicer | SICER | CONVERT_SICER GEM(ch_gem) CONVERT_SICER.out.peak From caa2ec230dee2845255481f3b7f435f467f64459 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 12:26:06 -0400 Subject: [PATCH 009/100] fix: ppqt accepts paired BAM see https://github.com/nf-core/modules/blob/29595ea381f4be8308c4af075eb64764c0249739/modules/nf-core/phantompeakqualtools/main.nf --- modules/local/qc.nf | 54 +++++++++++++++++++++------------------------ 1 file changed, 25 insertions(+), 29 deletions(-) diff --git a/modules/local/qc.nf b/modules/local/qc.nf index 1e5ff261..1f240cc1 100644 --- a/modules/local/qc.nf +++ b/modules/local/qc.nf @@ -157,42 +157,38 @@ process PHANTOM_PEAKS { def prefix = task.ext.prefix ?: "${meta.id}" def VERSION = '1.2.2' // WARN: Version information not provided by tool on CLI. Please update this string when bumping container versions. def prep_env = """ - # current working directory is a tmpdir when 'scratch' is set - TMP=tmp/ - mkdir \$TMP - trap 'rm -rf "\$TMP"' EXIT - # get spp path - RUN_SPP=\$(which run_spp.R) + """ def cleanup = """ - # get fragment length - frag_len=`cut -f 3 ${prefix}.spp.out | sed 's/,.*//g'` - echo \$frag_len > "${meta.id}.fraglen.txt" - # export versions - cat <<-END_VERSIONS > versions.yml - "${task.process}": - phantompeakqualtools: $VERSION - END_VERSIONS - """ - if (meta.single_end) { - """ - ${prep_env} - Rscript \$RUN_SPP -c=${bam} -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP - ${cleanup} """ - } else { // for PE, just use first read of each pair - """ - ${prep_env} - samtools view -b -f 66 -o \$TMP/${bam.baseName}.f66.bam ${bam} - samtools index \$TMP/${bam.baseName}.f66.bam - Rscript \$RUN_SPP -c=\$TMP/${bam.baseName}.f66.bam -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP - ${cleanup} - """ - } + """ + # current working directory is a tmpdir when 'scratch' is set + TMP=tmp/ + mkdir \$TMP + trap 'rm -rf "\$TMP"' EXIT + + Rscript \$(which run_spp.R) -c=${bam} -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP + + # get fragment length + frag_len=`cut -f 3 ${prefix}.spp.out | sed 's/,.*//g'` + echo \$frag_len > "${meta.id}.fraglen.txt" + + # export versions + cat <<-END_VERSIONS > versions.yml + "${task.process}": + phantompeakqualtools: $VERSION + END_VERSIONS + """ stub: """ touch ${meta.id}.ppqt.pdf ${meta.id}.spp.out "${meta.id}.fraglen.txt" + + # export versions + cat <<-END_VERSIONS > versions.yml + "${task.process}": + phantompeakqualtools: $VERSION + END_VERSIONS """ } From fa01f9185b5d02a103a61eeb4a49f162e0f587eb Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 12:26:34 -0400 Subject: [PATCH 010/100] chore: cleanup unused variables --- modules/local/qc.nf | 5 ----- 1 file changed, 5 deletions(-) diff --git a/modules/local/qc.nf b/modules/local/qc.nf index 1f240cc1..944e8929 100644 --- a/modules/local/qc.nf +++ b/modules/local/qc.nf @@ -156,11 +156,6 @@ process PHANTOM_PEAKS { script: def prefix = task.ext.prefix ?: "${meta.id}" def VERSION = '1.2.2' // WARN: Version information not provided by tool on CLI. Please update this string when bumping container versions. - def prep_env = """ - - """ - def cleanup = """ - """ """ # current working directory is a tmpdir when 'scratch' is set TMP=tmp/ From 36347d36b10057a0b70f7cc69a1c3f216768cc76 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 12:43:30 -0400 Subject: [PATCH 011/100] fix: bedtools output filename --- modules/local/bedtools.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/bedtools.nf b/modules/local/bedtools.nf index 4fe44c7d..914cce00 100644 --- a/modules/local/bedtools.nf +++ b/modules/local/bedtools.nf @@ -7,7 +7,7 @@ process BAM_TO_BED { tuple val(meta), path(bam) output: - tuple val(meta), path(bed), emit: bed + tuple val(meta), path("${bam.baseName}.bed"), emit: bed script: """ From 4079b3c25fcec246a5c74237590084a4866db4b9 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 12:43:47 -0400 Subject: [PATCH 012/100] fix: sicer channel doesn't need format --- subworkflows/local/peaks.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/subworkflows/local/peaks.nf b/subworkflows/local/peaks.nf index 7abec492..7128a515 100644 --- a/subworkflows/local/peaks.nf +++ b/subworkflows/local/peaks.nf @@ -57,8 +57,8 @@ workflow CALL_PEAKS { tag_all_bed .combine(tag_all_bed) .map { - meta1, tag1, format1, meta2, tag2, format2 -> - meta1.control == meta2.id && format1 == format2 ? [ meta1, tag1, tag2 ]: null + meta1, tag1, meta2, tag2 -> + meta1.control == meta2.id ? [ meta1, tag1, tag2 ]: null } .join(frag_lengths) .combine(genome_frac) From 27a90b496c575d2e5e45b96d3a8870cba10dea78 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 12:44:18 -0400 Subject: [PATCH 013/100] feat: create test samplesheet w/ mixed single & paired end --- assets/samplesheet_test_paired.csv | 7 +++++++ conf/test_paired.config | 6 +++--- 2 files changed, 10 insertions(+), 3 deletions(-) create mode 100644 assets/samplesheet_test_paired.csv diff --git a/assets/samplesheet_test_paired.csv b/assets/samplesheet_test_paired.csv new file mode 100644 index 00000000..58a50b71 --- /dev/null +++ b/assets/samplesheet_test_paired.csv @@ -0,0 +1,7 @@ +sample,fastq_1,fastq_2,antibody,control +SPT5_T0_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822153_1.fastq.gz,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822153_2.fastq.gz,SPT5,SPT5_INPUT_REP1 +SPT5_T0_REP2,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822154_1.fastq.gz,,SPT5,SPT5_INPUT_REP2 +SPT5_T15_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822157_1.fastq.gz,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822157_2.fastq.gz,SPT5,SPT5_INPUT_REP1 +SPT5_T15_REP2,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822158_1.fastq.gz,,SPT5,SPT5_INPUT_REP2 +SPT5_INPUT_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/testdata/SRR5204809_Spt5-ChIP_Input1_SacCer_ChIP-Seq_ss100k_R1.fastq.gz,https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/testdata/SRR5204809_Spt5-ChIP_Input1_SacCer_ChIP-Seq_ss100k_R2.fastq.gz,, +SPT5_INPUT_REP2,https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/testdata/SRR5204810_Spt5-ChIP_Input2_SacCer_ChIP-Seq_ss100k_R1.fastq.gz,,, diff --git a/conf/test_paired.config b/conf/test_paired.config index 5f616304..395d282a 100644 --- a/conf/test_paired.config +++ b/conf/test_paired.config @@ -1,9 +1,9 @@ params { - config_profile_name = 'Test paired-end' - config_profile_description = 'Minimal paired-end test dataset to check pipeline function' + config_profile_name = 'Test paired-end and single-end mixed' + config_profile_description = 'Minimal test dataset to check paired & single end handling' outdir = 'results/test_paired' - input = 'https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/samplesheet/v2.0/samplesheet_test.csv' + input = 'assets/samplesheet_test_paired.csv' // adapted from 'https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/samplesheet/v2.0/samplesheet_test.csv' genome = 'hg38' max_cpus = 32 // for interactive node on biowulf From 91190833558bb813857ae844d76bd0abe97bb8a0 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 12:55:24 -0400 Subject: [PATCH 014/100] fix: output dedup flagstat for qc table --- subworkflows/local/deduplicate.nf | 5 ++++- 1 file changed, 4 insertions(+), 1 deletion(-) diff --git a/subworkflows/local/deduplicate.nf b/subworkflows/local/deduplicate.nf index 35d660cd..87f2371c 100644 --- a/subworkflows/local/deduplicate.nf +++ b/subworkflows/local/deduplicate.nf @@ -21,7 +21,9 @@ workflow DEDUPLICATE { INDEX_PAIRED.out.bam .mix(INDEX_SINGLE.out.bam) .set{ ch_bam_bai } - + INDEX_PAIRED.out.flagstat + .mix(INDEX_SINGLE.out.flagstat) + .set{ ch_flagstat } // mix single bed and paired bam channel for peak callers, with variable to designate format INDEX_PAIRED.out.bam .map{ meta, bam, bai -> @@ -34,4 +36,5 @@ workflow DEDUPLICATE { emit: bam = ch_bam_bai tag_align = mixed_tagAlign + flagstat = ch_flagstat } From dbcdc329f0232f124f07cc0d9f34353bf0a80018 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 12:56:25 -0400 Subject: [PATCH 015/100] feat: support paired end in deeptools fingerprint & bamCoverage --- modules/local/deeptools.nf | 26 ++++++++++++++------------ 1 file changed, 14 insertions(+), 12 deletions(-) diff --git a/modules/local/deeptools.nf b/modules/local/deeptools.nf index baff9d4e..bb05f900 100644 --- a/modules/local/deeptools.nf +++ b/modules/local/deeptools.nf @@ -15,18 +15,19 @@ process BAM_COVERAGE { val(meta), emit: meta path("${meta.id}.bw"), emit: bigwig - script: // https://deeptools.readthedocs.io/en/2.1.0/content/tools/bamCoverage.html + script: + def args = meta.single_end ? "--extendReads ${fraglen}" : '--centerReads' """ - bamCoverage \ - --bam ${bam} \ - -o ${meta.id}.bw \ - --binSize ${params.deeptools.bin_size} \ - --smoothLength ${params.deeptools.smooth_length} \ - --ignoreForNormalization ${params.deeptools.excluded_chroms} \ - --numberOfProcessors ${task.cpus} \ - --normalizeUsing ${params.deeptools.normalize_using} \ - --effectiveGenomeSize ${params.genomes[ params.genome ].effective_genome_size} \ - --extendReads ${fraglen} + bamCoverage \\ + --bam ${bam} \\ + -o ${meta.id}.bw \\ + --binSize ${params.deeptools.bin_size} \\ + --smoothLength ${params.deeptools.smooth_length} \\ + --ignoreForNormalization ${params.deeptools.excluded_chroms} \\ + --numberOfProcessors ${task.cpus} \\ + --normalizeUsing ${params.deeptools.normalize_using} \\ + --effectiveGenomeSize ${params.genomes[ params.genome ].effective_genome_size} \\ + ${args} """ stub: @@ -140,6 +141,7 @@ process PLOT_FINGERPRINT { script: // TODO handle extendReads for single vs paired https://github.com/nf-core/chipseq/blob/51eba00b32885c4d0bec60db3cb0a45eb61e34c5/modules/nf-core/modules/deeptools/plotfingerprint/main.nf def prefix = task.ext.prefix ?: "${meta.id}" + def args = meta.single_end ? '--extendReads 200' : '' """ plotFingerprint \\ --bamfiles ${bams.join(' ')} \\ @@ -149,7 +151,7 @@ process PLOT_FINGERPRINT { --numberOfProcessors ${task.cpus} \\ --skipZeros \\ --smartLabels \\ - --extendReads 200 + ${args} """ stub: From 5df4ea414f9ab2a3a8fa1379c1739f43ecc09cd5 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 13:37:51 -0400 Subject: [PATCH 016/100] fix: don't use getBaseName(2) in fastqc breaks for paired end --- modules/local/qc.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/modules/local/qc.nf b/modules/local/qc.nf index 944e8929..66597735 100644 --- a/modules/local/qc.nf +++ b/modules/local/qc.nf @@ -10,8 +10,8 @@ process FASTQC { input: tuple val(meta), path(fastq), val(fqtype) output: - path("${fastq.getBaseName(2)}*.html"), emit: html - path ("${fastq.getBaseName(2)}*.zip"), emit: zip + path("*.html"), emit: html + path ("*.zip"), emit: zip script: """ @@ -23,7 +23,7 @@ process FASTQC { stub: """ - touch ${fastq.getBaseName(2)}_fastqc.html ${fastq.getBaseName(2)}_fastqc.zip + touch ${meta.id}_fastqc.html ${meta.id}_fastqc.zip """ } From 8bf1ff6ef5a1ddceb306964c2d5ac8f1311cbe54 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 13:39:03 -0400 Subject: [PATCH 017/100] fix: Revert "fix: ppqt accepts paired BAM" we do need to use -f 66 with ppqt. will need to fix samtools view -q 6 for bam with vishal's custom script This reverts commit caa2ec230dee2845255481f3b7f435f467f64459. --- modules/local/qc.nf | 57 ++++++++++++++++++++++++++------------------- 1 file changed, 33 insertions(+), 24 deletions(-) diff --git a/modules/local/qc.nf b/modules/local/qc.nf index 66597735..3da57228 100644 --- a/modules/local/qc.nf +++ b/modules/local/qc.nf @@ -156,34 +156,43 @@ process PHANTOM_PEAKS { script: def prefix = task.ext.prefix ?: "${meta.id}" def VERSION = '1.2.2' // WARN: Version information not provided by tool on CLI. Please update this string when bumping container versions. - """ - # current working directory is a tmpdir when 'scratch' is set - TMP=tmp/ - mkdir \$TMP - trap 'rm -rf "\$TMP"' EXIT - - Rscript \$(which run_spp.R) -c=${bam} -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP - - # get fragment length - frag_len=`cut -f 3 ${prefix}.spp.out | sed 's/,.*//g'` - echo \$frag_len > "${meta.id}.fraglen.txt" - - # export versions - cat <<-END_VERSIONS > versions.yml - "${task.process}": - phantompeakqualtools: $VERSION - END_VERSIONS - """ + def prep_env = """ + # current working directory is a tmpdir when 'scratch' is set + TMP=tmp/ + mkdir \$TMP + trap 'rm -rf "\$TMP"' EXIT + # get spp path + RUN_SPP=\$(which run_spp.R) + """ + def cleanup = """ + # get fragment length + frag_len=`cut -f 3 ${prefix}.spp.out | sed 's/,.*//g'` + echo \$frag_len > "${meta.id}.fraglen.txt" + # export versions + cat <<-END_VERSIONS > versions.yml + "${task.process}": + phantompeakqualtools: $VERSION + END_VERSIONS + """ + if (meta.single_end) { + """ + ${prep_env} + Rscript \$RUN_SPP -c=${bam} -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP + ${cleanup} + """ + } else { // for PE, just use first read of each pair + """ + ${prep_env} + samtools view -b -f 66 -o \$TMP/${bam.baseName}.f66.bam ${bam} + samtools index \$TMP/${bam.baseName}.f66.bam + Rscript \$RUN_SPP -c=\$TMP/${bam.baseName}.f66.bam -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP + ${cleanup} + """ + } stub: """ touch ${meta.id}.ppqt.pdf ${meta.id}.spp.out "${meta.id}.fraglen.txt" - - # export versions - cat <<-END_VERSIONS > versions.yml - "${task.process}": - phantompeakqualtools: $VERSION - END_VERSIONS """ } From 94271607d950b5d1b8195f69ab0107bd425a0fe1 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 13:45:39 -0400 Subject: [PATCH 018/100] feat: add special bam PE filter script from old pipeliner source https://github.com/CCBR/Pipeliner/blob/86c6ccaa3d58381a0ffd696bbf9c047e4f991f9e/Results-template/Scripts/bam_filter_by_mapq.py --- bin/bam_filter_by_mapq.py | 51 +++++++++++++++++++++++++++++++++++++++ 1 file changed, 51 insertions(+) create mode 100755 bin/bam_filter_by_mapq.py diff --git a/bin/bam_filter_by_mapq.py b/bin/bam_filter_by_mapq.py new file mode 100755 index 00000000..ef345084 --- /dev/null +++ b/bin/bam_filter_by_mapq.py @@ -0,0 +1,51 @@ +#!/usr/bin/env python + +""" +source https://github.com/CCBR/Pipeliner/blob/86c6ccaa3d58381a0ffd696bbf9c047e4f991f9e/Results-template/Scripts/bam_filter_by_mapq.py +""" + +import pysam, sys +import argparse + +parser = argparse.ArgumentParser(description="filter PE bamfile by mapQ values") +parser.add_argument("-i", dest="inBam", required=True, help="Input Bam File") +parser.add_argument("-o", dest="outBam", required=True, help="Output Bam File") +parser.add_argument( + "-q", + dest="mapQ", + type=int, + required=False, + help="mapQ value ... default 6", + default=6, +) +args = parser.parse_args() +samfile = pysam.AlignmentFile(args.inBam, "rb") +mapq = dict() +for read in samfile.fetch(): + if read.is_unmapped: + continue + if read.is_supplementary: + continue + if read.is_secondary: + continue + if read.is_duplicate: + continue + if read.is_proper_pair: + if read.mapping_quality < args.mapQ and read.query_name in mapq: + del mapq[read.query_name] + if read.mapping_quality >= args.mapQ and not read.query_name in mapq: + mapq[read.query_name] = 1 +samfile.close() +samfile = pysam.AlignmentFile(args.inBam, "rb") +pairedreads = pysam.AlignmentFile(args.outBam, "wb", template=samfile) +for read in samfile.fetch(): + if read.query_name in mapq: + if read.is_supplementary: + continue + if read.is_secondary: + continue + if read.is_duplicate: + continue + pairedreads.write(read) +samfile.close() +pairedreads.close() From 3c06d393e7d5796000964fc6a0ec2d975e70d822 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 15:11:37 -0400 Subject: [PATCH 019/100] fix: use picard for bam_to_fastq when paired reads --- modules/local/align.nf | 65 +++++++++++++++++++++++++++++++++++------- 1 file changed, 54 insertions(+), 11 deletions(-) diff --git a/modules/local/align.nf b/modules/local/align.nf index c91bb9b8..3241347c 100644 --- a/modules/local/align.nf +++ b/modules/local/align.nf @@ -11,25 +11,56 @@ process ALIGN_BLACKLIST { path(blacklist_files) output: - tuple val(meta), path("${meta.id}.no_blacklist.fastq.gz"), emit: reads + tuple val(meta), path("${meta.id}.no_blacklist.bam"), emit: bam script: def prefix = task.ext.prefix ?: "${meta.id}" def filter_flag = meta.single_end ? '4' : '12' """ - bwa mem -t $task.cpus ${params.genomes[ params.genome ].blacklist} $fastq > ${prefix}.sam + bwa mem -t $task.cpus ${params.genomes[ params.genome ].blacklist} $fastq > ${prefix}.bam samtools view \\ -@ ${task.cpus} \\ -f${filter_flag} \\ -b \\ - ${prefix}.sam |\ - samtools bam2fq |\ - pigz -p $task.cpus > ${prefix}.no_blacklist.fastq.gz + ${prefix}.bam > ${meta.id}.no_blacklist.bam """ stub: """ - touch ${meta.id}.no_blacklist.fastq.gz + touch ${meta.id}.no_blacklist.bam + """ +} + +process BAM_TO_FASTQ { + tag { meta.id } + label 'align' + container "${ meta.single_end ? params.containers.base : params.containers.picard }" + + input: + tuple val(meta), path(bam) + + output: + tuple val(meta), path("*.R?.fastq*"), emit: reads + + script: + if (meta.single_end) { + """ + samtools bam2fq ${bam} | pigz -p ${task.cpus} > ${bam.baseName}.R1.fastq.gz + """ + } else { + """ + picard -Xmx${task.memory.toGiga()}G SamToFastq \\ + --VALIDATION_STRINGENCY SILENT \\ + --INPUT ${bam} \\ + --FASTQ ${bam.baseName}.R1.fastq \\ + --SECOND_END_FASTQ ${bam.baseName}.R2.fastq \\ + --UNPAIRED_FASTQ ${bam.baseName}.unpaired.fastq + # TODO: gzip output. need container w/ both picard & pigz + """ + } + stub: + """ + touch ${bam.baseName}.fastq """ } @@ -50,6 +81,21 @@ process ALIGN_GENOME { script: def prefix = task.ext.prefix ?: "${meta.id}" + def filter = '' + if (meta.single_end) { + filter = """samtools view \\ + -@ ${task.cpus} \\ + -q ${params.align_min_quality} \\ + -b \\ + ${prefix}.sorted.bam > ${prefix}.aligned.filtered.bam + """ + } else { + filter = """bam_filter_by_mapq.py \\ + -q ${params.align_min_quality} \\ + -i ${prefix}.sorted.bam \\ + -o ${prefix}.aligned.filtered.bam + """ + } """ # current working directory is a tmpdir when 'scratch' is set TMP=tmp/ @@ -62,11 +108,8 @@ process ALIGN_GENOME { -m 2G \\ -T \$TMP \\ ${prefix}.bam > ${prefix}.sorted.bam - samtools view \\ - -@ ${task.cpus} \\ - -q ${params.align_min_quality} \\ - -b \\ - ${prefix}.sorted.bam > ${prefix}.aligned.filtered.bam + samtools index ${prefix}.sorted.bam + ${filter} samtools flagstat ${prefix}.aligned.filtered.bam > ${prefix}.aligned.filtered.bam.flagstat """ From d1e2f2a008607901f3b83a1080888273704ac0c4 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 15:12:14 -0400 Subject: [PATCH 020/100] style: don't need equals sign in process container def --- modules/local/deduplicate.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/deduplicate.nf b/modules/local/deduplicate.nf index 7beb369e..7ae8368e 100644 --- a/modules/local/deduplicate.nf +++ b/modules/local/deduplicate.nf @@ -4,7 +4,7 @@ process MACS2_DEDUP { label 'dedup' label 'process_medium' - container = "${params.containers.macs2}" + container "${params.containers.macs2}" input: tuple val(meta), path(bam), path(chrom_sizes) @@ -46,7 +46,7 @@ process PICARD_DEDUP { label 'dedup' label 'process_medium' - container = "${params.containers.picard}" + container "${params.containers.picard}" input: tuple val(meta), path(bam) From 31c4ff6a6fbc89f19d3e670bff89d49e7f7943be Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 15:12:44 -0400 Subject: [PATCH 021/100] fix: use BAM_TO_FASTQ --- main.nf | 9 +++++---- 1 file changed, 5 insertions(+), 4 deletions(-) diff --git a/main.nf b/main.nf index e03dc482..63a74075 100644 --- a/main.nf +++ b/main.nf @@ -25,8 +25,9 @@ include { CALL_PEAKS } from './subworkflows/local/peaks.nf' // MODULES include { CUTADAPT } from "./modules/CCBR/cutadapt" -include { ALIGN_BLACKLIST } from "./modules/local/align.nf" -include { ALIGN_GENOME } from "./modules/local/align.nf" +include { ALIGN_BLACKLIST + BAM_TO_FASTQ + ALIGN_GENOME } from "./modules/local/align.nf" include { PHANTOM_PEAKS } from "./modules/local/qc.nf" include { PPQT_PROCESS MULTIQC } from "./modules/local/qc.nf" @@ -42,11 +43,11 @@ workflow { Channel.fromPath(params.genomes[ params.genome ].blacklist_files, checkIfExists: true) .collect() .set{ blacklist_files } - ALIGN_BLACKLIST(trimmed_fastqs, blacklist_files) + ALIGN_BLACKLIST(trimmed_fastqs, blacklist_files) | BAM_TO_FASTQ Channel.fromPath(params.genomes[ params.genome ].reference_files, checkIfExists: true) .collect() .set{ reference_files } - ALIGN_GENOME(ALIGN_BLACKLIST.out.reads, reference_files) + ALIGN_GENOME(BAM_TO_FASTQ.out.reads, reference_files) ALIGN_GENOME.out.bam.set{ aligned_bam } Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) From 93f9b421480db29939cd0ea1b0b8a010ceddbb70 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 29 Sep 2023 15:53:11 -0400 Subject: [PATCH 022/100] refactor: simplify logic for ppqt bam preparation --- modules/local/qc.nf | 48 +++++++++++++++++++-------------------------- 1 file changed, 20 insertions(+), 28 deletions(-) diff --git a/modules/local/qc.nf b/modules/local/qc.nf index 3da57228..7958656a 100644 --- a/modules/local/qc.nf +++ b/modules/local/qc.nf @@ -156,39 +156,31 @@ process PHANTOM_PEAKS { script: def prefix = task.ext.prefix ?: "${meta.id}" def VERSION = '1.2.2' // WARN: Version information not provided by tool on CLI. Please update this string when bumping container versions. - def prep_env = """ - # current working directory is a tmpdir when 'scratch' is set - TMP=tmp/ - mkdir \$TMP - trap 'rm -rf "\$TMP"' EXIT - # get spp path - RUN_SPP=\$(which run_spp.R) - """ - def cleanup = """ - # get fragment length - frag_len=`cut -f 3 ${prefix}.spp.out | sed 's/,.*//g'` - echo \$frag_len > "${meta.id}.fraglen.txt" - # export versions - cat <<-END_VERSIONS > versions.yml - "${task.process}": - phantompeakqualtools: $VERSION - END_VERSIONS - """ + def prep_bam = '' if (meta.single_end) { - """ - ${prep_env} - Rscript \$RUN_SPP -c=${bam} -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP - ${cleanup} + prep_bam = """ + BAM=${bam} """ } else { // for PE, just use first read of each pair - """ - ${prep_env} - samtools view -b -f 66 -o \$TMP/${bam.baseName}.f66.bam ${bam} - samtools index \$TMP/${bam.baseName}.f66.bam - Rscript \$RUN_SPP -c=\$TMP/${bam.baseName}.f66.bam -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out -tmpdir=\$TMP - ${cleanup} + prep_bam = """ + BAM=${bam.baseName}.f66.bam + samtools view -b -f 64 -o \$BAM ${bam} + samtools index \$BAM """ } + """ + ${prep_bam} + Rscript \$(which run_spp.R) -c=\$BAM -savp=${prefix}.ppqt.pdf -out=${prefix}.spp.out + # get fragment length + frag_len=`cut -f 3 ${prefix}.spp.out | sed 's/,.*//g'` + echo \$frag_len > "${meta.id}.fraglen.txt" + + # export versions + cat <<-END_VERSIONS > versions.yml + "${task.process}": + phantompeakqualtools: $VERSION + END_VERSIONS + """ stub: """ From 247e4fe215496180c75bd47d11ed4b6a3fdbc38b Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 2 Oct 2023 10:58:51 -0400 Subject: [PATCH 023/100] refactor: get index files from PREPARE_GENOME subworkflow [WIP] --- main.nf | 22 ++++++++-------------- modules/local/align.nf | 4 +++- nextflow.config | 3 ++- subworkflows/local/prepare_genome.nf | 21 +++++++++++++++++++-- 4 files changed, 32 insertions(+), 18 deletions(-) diff --git a/main.nf b/main.nf index b46f4024..e1580c88 100644 --- a/main.nf +++ b/main.nf @@ -18,9 +18,10 @@ log.info """\ .stripIndent() // SUBWORKFLOWS -include { INPUT_CHECK } from './subworkflows/local/input_check.nf' -include { QC } from './subworkflows/local/qc.nf' -include { CALL_PEAKS } from './subworkflows/local/peaks.nf' +include { INPUT_CHECK } from './subworkflows/local/input_check.nf' +include { PREPARE_GENOME } from './subworkflows/local/prepare_genome.nf' +include { QC } from './subworkflows/local/qc.nf' +include { CALL_PEAKS } from './subworkflows/local/peaks.nf' // MODULES include { TRIM_SE } from "./modules/local/trim.nf" @@ -38,19 +39,12 @@ workflow { raw_fastqs | TRIM_SE TRIM_SE.out.set{ trimmed_fastqs } - Channel.fromPath(params.genomes[ params.genome ].blacklist_files, checkIfExists: true) - .collect() - .set{ blacklist_files } - ALIGN_BLACKLIST(trimmed_fastqs, blacklist_files) - Channel.fromPath(params.genomes[ params.genome ].reference_files, checkIfExists: true) - .collect() - .set{ reference_files } - ALIGN_GENOME(ALIGN_BLACKLIST.out.reads, reference_files) + PREPARE_GENOME() + ALIGN_BLACKLIST(trimmed_fastqs, PREPARE_GENOME.out.blacklist_files) + ALIGN_GENOME(ALIGN_BLACKLIST.out.reads, PREPARE_GENOME.out.reference_files) ALIGN_GENOME.out.bam.set{ aligned_bam } - Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) - .set{ chrom_sizes } - aligned_bam.combine(chrom_sizes) | DEDUPLICATE + aligned_bam.combine(PREPARE_GENOME.out.chrom_sizes) | DEDUPLICATE DEDUPLICATE.out.bam.set{ deduped_bam } DEDUPLICATE.out.tag_align.set{ deduped_tagalign } diff --git a/modules/local/align.nf b/modules/local/align.nf index 69598378..0217f6fd 100644 --- a/modules/local/align.nf +++ b/modules/local/align.nf @@ -16,7 +16,9 @@ process ALIGN_BLACKLIST { script: // TODO use samtools -f4 for single-end and -f12 for paired to get unmapped reads https://broadinstitute.github.io/picard/explain-flags.html def prefix = task.ext.prefix ?: "${meta.id}" """ - bwa mem -t $task.cpus ${params.genomes[ params.genome ].blacklist} $fastq > ${prefix}.sam + INDEX=`find -L ./ -name "*.amb" | sed 's/.amb//' | sed 's|^./||'` + + bwa mem -t $task.cpus \$INDEX $fastq > ${prefix}.sam samtools view \\ -@ ${task.cpus} \\ -f4 \\ diff --git a/nextflow.config b/nextflow.config index 64962ba0..7580f0e8 100644 --- a/nextflow.config +++ b/nextflow.config @@ -4,7 +4,8 @@ params { input = null seq_center = null genome = null - genome_base = null + fasta = null + gtf = null // Boilerplate options outdir = 'results' diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index b465acaa..8e9564ea 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -1,4 +1,21 @@ workflow PREPARE_GENOME { - ch_fasta = file(params.fasta) - ch_gtf = file(params.fasta) + main: + if (params.fasta && params.gtf) { + ch_fasta = file(params.fasta) + ch_gtf = file(params.gtf) + } else { + Channel.fromPath(params.genomes[ params.genome ].blacklist_files, checkIfExists: true) + .collect() + .set{ ch_blacklist_files } + Channel.fromPath(params.genomes[ params.genome ].reference_files, checkIfExists: true) + .collect() + .set{ ch_reference_files } + Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) + .set{ ch_chrom_sizes } + } + + emit: + blacklist_files = ch_blacklist_files + reference_files = ch_reference_files + chrom_sizes = ch_chrom_sizes } From 8b5fd5823e3ef78dfe27361486c88e7caefe0ca2 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 2 Oct 2023 11:11:57 -0400 Subject: [PATCH 024/100] fix: explicitly pass blacklist index name --- main.nf | 2 +- modules/local/align.nf | 7 +++---- subworkflows/local/prepare_genome.nf | 1 + 3 files changed, 5 insertions(+), 5 deletions(-) diff --git a/main.nf b/main.nf index e1580c88..aab7ce11 100644 --- a/main.nf +++ b/main.nf @@ -40,7 +40,7 @@ workflow { TRIM_SE.out.set{ trimmed_fastqs } PREPARE_GENOME() - ALIGN_BLACKLIST(trimmed_fastqs, PREPARE_GENOME.out.blacklist_files) + ALIGN_BLACKLIST(trimmed_fastqs, PREPARE_GENOME.out.blacklist_files, PREPARE_GENOME.out.blacklist_name) ALIGN_GENOME(ALIGN_BLACKLIST.out.reads, PREPARE_GENOME.out.reference_files) ALIGN_GENOME.out.bam.set{ aligned_bam } diff --git a/modules/local/align.nf b/modules/local/align.nf index 0217f6fd..ff5e4dbb 100644 --- a/modules/local/align.nf +++ b/modules/local/align.nf @@ -8,7 +8,8 @@ process ALIGN_BLACKLIST { input: tuple val(meta), path(fastq) - path(blacklist_files) + path(index_files) + val(index_name) output: tuple val(meta), path("${meta.id}.no_blacklist.fastq.gz"), emit: reads @@ -16,9 +17,7 @@ process ALIGN_BLACKLIST { script: // TODO use samtools -f4 for single-end and -f12 for paired to get unmapped reads https://broadinstitute.github.io/picard/explain-flags.html def prefix = task.ext.prefix ?: "${meta.id}" """ - INDEX=`find -L ./ -name "*.amb" | sed 's/.amb//' | sed 's|^./||'` - - bwa mem -t $task.cpus \$INDEX $fastq > ${prefix}.sam + bwa mem -t ${task.cpus} ${index_name} ${fastq} > ${prefix}.sam samtools view \\ -@ ${task.cpus} \\ -f4 \\ diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index 8e9564ea..8e91b53d 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -16,6 +16,7 @@ workflow PREPARE_GENOME { emit: blacklist_files = ch_blacklist_files + blacklist_name = Channel.value(params.genomes[ params.genome ].blacklist) reference_files = ch_reference_files chrom_sizes = ch_chrom_sizes } From bce5ba4b4149b4ec7a4077b750654e9bebc27efa Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 2 Oct 2023 11:14:55 -0400 Subject: [PATCH 025/100] refactor: use ternary operator for blacklist name --- subworkflows/local/prepare_genome.nf | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index 8e91b53d..ea45b533 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -1,5 +1,7 @@ workflow PREPARE_GENOME { main: + ch_blacklist_name = params.genomes[ params.genome ].blacklist ? Channel.value(params.genomes[ params.genome ].blacklist) : "${params.genome}.blacklist" + if (params.fasta && params.gtf) { ch_fasta = file(params.fasta) ch_gtf = file(params.gtf) @@ -16,7 +18,7 @@ workflow PREPARE_GENOME { emit: blacklist_files = ch_blacklist_files - blacklist_name = Channel.value(params.genomes[ params.genome ].blacklist) + blacklist_name = ch_blacklist_name reference_files = ch_reference_files chrom_sizes = ch_chrom_sizes } From f1304ad6d0b2dfd7f1c15486f985accd995661e4 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 2 Oct 2023 13:31:02 -0400 Subject: [PATCH 026/100] refactor: get effective genome size from PREP_GENOME subwf --- main.nf | 9 ++++++--- modules/local/deeptools.nf | 5 ++--- modules/local/peaks.nf | 11 ++++++----- modules/local/qc.nf | 3 ++- subworkflows/local/peaks.nf | 3 ++- subworkflows/local/prepare_genome.nf | 7 +++++++ subworkflows/local/qc.nf | 7 ++++--- 7 files changed, 29 insertions(+), 16 deletions(-) diff --git a/main.nf b/main.nf index aab7ce11..3c6456f4 100644 --- a/main.nf +++ b/main.nf @@ -40,11 +40,12 @@ workflow { TRIM_SE.out.set{ trimmed_fastqs } PREPARE_GENOME() + effective_genome_size = PREPARE_GENOME.out.effective_genome_size ALIGN_BLACKLIST(trimmed_fastqs, PREPARE_GENOME.out.blacklist_files, PREPARE_GENOME.out.blacklist_name) ALIGN_GENOME(ALIGN_BLACKLIST.out.reads, PREPARE_GENOME.out.reference_files) ALIGN_GENOME.out.bam.set{ aligned_bam } - aligned_bam.combine(PREPARE_GENOME.out.chrom_sizes) | DEDUPLICATE + DEDUPLICATE(aligned_bam.combine(PREPARE_GENOME.out.chrom_sizes), effective_genome_size) DEDUPLICATE.out.bam.set{ deduped_bam } DEDUPLICATE.out.tag_align.set{ deduped_tagalign } @@ -56,7 +57,9 @@ workflow { QC(raw_fastqs, trimmed_fastqs, aligned_bam, ALIGN_GENOME.out.flagstat, deduped_bam, DEDUPLICATE.out.flagstat, - PHANTOM_PEAKS.out.spp, frag_lengths + PHANTOM_PEAKS.out.spp, frag_lengths, + PREPARE_GENOME.out.gene_info, + effective_genome_size ) QC.out.bigwigs.set{ ch_ip_ctrl_bigwig } @@ -66,6 +69,6 @@ workflow { } if (params.run.call_peaks) { - CALL_PEAKS(chrom_sizes, deduped_tagalign, frag_lengths) + CALL_PEAKS(chrom_sizes, deduped_tagalign, frag_lengths, effective_genome_size) } } diff --git a/modules/local/deeptools.nf b/modules/local/deeptools.nf index baff9d4e..c9e7d38b 100644 --- a/modules/local/deeptools.nf +++ b/modules/local/deeptools.nf @@ -8,8 +8,7 @@ process BAM_COVERAGE { container = "${params.containers.deeptools}" input: - tuple val(meta), path(bam), path(bai) - tuple val(meta), val(fraglen) + tuple val(meta), path(bam), path(bai), val(fraglen), val(effective_genome_size) output: val(meta), emit: meta @@ -25,7 +24,7 @@ process BAM_COVERAGE { --ignoreForNormalization ${params.deeptools.excluded_chroms} \ --numberOfProcessors ${task.cpus} \ --normalizeUsing ${params.deeptools.normalize_using} \ - --effectiveGenomeSize ${params.genomes[ params.genome ].effective_genome_size} \ + --effectiveGenomeSize ${effective_genome_size} \ --extendReads ${fraglen} """ diff --git a/modules/local/peaks.nf b/modules/local/peaks.nf index 0f8b3909..27cbe0af 100644 --- a/modules/local/peaks.nf +++ b/modules/local/peaks.nf @@ -6,13 +6,14 @@ process CALC_GENOME_FRAC { input: path(chrom_sizes) + val(effective_genome_size) output: env(genome_frac), emit: genome_frac script: """ - genome_frac=`calc_effective_genome_fraction.py ${params.genomes[ params.genome ].effective_genome_size} ${chrom_sizes} ${params.deeptools.excluded_chroms}` + genome_frac=`calc_effective_genome_fraction.py effective_genome_size ${chrom_sizes} ${params.deeptools.excluded_chroms}` echo \$genome_frac """ @@ -31,7 +32,7 @@ process MACS_BROAD { container = "${params.containers.macs2}" input: - tuple val(meta), path(chip), path(input), val(fraglen), val(genome_frac) + tuple val(meta), path(chip), path(input), val(fraglen), val(genome_frac), val(effective_genome_size) output: path("${meta.id}_peaks.xls") @@ -43,7 +44,7 @@ process MACS_BROAD { macs2 callpeak \\ -t ${chip} \\ -c ${input} \\ - -g ${params.genomes[ params.genome ].effective_genome_size} \\ + -g ${effective_genome_size} \\ -n ${meta.id} \\ --extsize ${fraglen} \\ --nomodel \\ @@ -69,7 +70,7 @@ process MACS_NARROW { container = "${params.containers.macs2}" input: - tuple val(meta), path(chip), path(input), val(fraglen), val(genome_frac) + tuple val(meta), path(chip), path(input), val(fraglen), val(genome_frac), val(effective_genome_size) output: path("${meta.id}_peaks.xls") @@ -81,7 +82,7 @@ process MACS_NARROW { macs2 callpeak \\ -t ${chip} \\ -c ${input} \\ - -g ${params.genomes[ params.genome ].effective_genome_size} \\ + -g ${effective_genome_size} \\ -n ${meta.id} \\ --extsize ${fraglen} \\ --nomodel \\ diff --git a/modules/local/qc.nf b/modules/local/qc.nf index c80fc1ed..7e69f0d2 100644 --- a/modules/local/qc.nf +++ b/modules/local/qc.nf @@ -202,6 +202,7 @@ process DEDUPLICATE { input: tuple val(meta), path(bam), path(chrom_sizes) + val(effective_genome_size) output: tuple val(meta), path("${meta.id}.TagAlign.bed"), emit: tag_align @@ -215,7 +216,7 @@ process DEDUPLICATE { mkdir \$TMP trap 'rm -rf "\$TMP"' EXIT - macs2 filterdup -i ${bam} -g ${params.genomes[ params.genome ].effective_genome_size} --keep-dup="auto" -o TmpTagAlign1 + macs2 filterdup -i ${bam} -g ${effective_genome_size} --keep-dup="auto" -o TmpTagAlign1 awk -F"\\t" -v OFS="\\t" '{{if (\$2>0 && \$3>0) {{print}}}}' TmpTagAlign1 > TmpTagAlign2 awk -F"\\t" -v OFS="\\t" '{{print \$1,1,\$2}}' ${chrom_sizes} > \$TMP/GenomeFile_unsorted.bed sort \\ diff --git a/subworkflows/local/peaks.nf b/subworkflows/local/peaks.nf index 04759788..1210a97f 100644 --- a/subworkflows/local/peaks.nf +++ b/subworkflows/local/peaks.nf @@ -11,10 +11,11 @@ workflow CALL_PEAKS { chrom_sizes deduped_tagalign frag_lengths + effective_genome_size main: // peak calling - genome_frac = CALC_GENOME_FRAC(chrom_sizes) + genome_frac = CALC_GENOME_FRAC(chrom_sizes, effective_genome_size) // create channel with [ meta, chip_tag, input_tag, fraglen, genome_frac] deduped_tagalign .combine(deduped_tagalign) diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index ea45b533..e077007d 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -14,6 +14,11 @@ workflow PREPARE_GENOME { .set{ ch_reference_files } Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) .set{ ch_chrom_sizes } + Channel.fromPath(params.genomes[ params.genome ].gene_info, + checkIfExists: true) + .set{ ch_gene_info } + Channel.value(params.genomes[ params.genome ].effective_genome_size) + .set{ ch_egs} } emit: @@ -21,4 +26,6 @@ workflow PREPARE_GENOME { blacklist_name = ch_blacklist_name reference_files = ch_reference_files chrom_sizes = ch_chrom_sizes + gene_info = ch_gene_info + effective_genome_size = ch_egs } diff --git a/subworkflows/local/qc.nf b/subworkflows/local/qc.nf index 946a8aea..099cf4c7 100644 --- a/subworkflows/local/qc.nf +++ b/subworkflows/local/qc.nf @@ -29,6 +29,8 @@ workflow QC { deduped_flagstat ppqt_spp frag_lengths + gene_info + effective_genome_size main: raw_fastqs.combine(Channel.value("raw")) | FASTQC_RAW @@ -66,7 +68,7 @@ workflow QC { // Deeptools - BAM_COVERAGE(deduped_bam, frag_lengths) + deduped_bam.join(frag_lengths).combine(effective_genome_size) | BAM_COVERAGE BAM_COVERAGE.out.bigwig.collect().set{ bigwig_list } BIGWIG_SUM(bigwig_list) BIGWIG_SUM.out.array.combine(Channel.from('heatmap', 'scatterplot')) | PLOT_CORRELATION @@ -81,8 +83,7 @@ workflow QC { } .set { ch_ip_ctrl_bam_bai } ch_ip_ctrl_bam_bai | PLOT_FINGERPRINT - Channel.fromPath(params.genomes[ params.genome ].gene_info, - checkIfExists: true) | BED_PROTEIN_CODING + gene_info | BED_PROTEIN_CODING COMPUTE_MATRIX(bigwig_list, BED_PROTEIN_CODING.out.bed.combine(Channel.from('metagene','TSS')) ) From e020e277244f47c5a5906512e31dbfd632301858 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 2 Oct 2023 13:34:32 -0400 Subject: [PATCH 027/100] refactor: macs accepts efg as input --- subworkflows/local/peaks.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/subworkflows/local/peaks.nf b/subworkflows/local/peaks.nf index 1210a97f..739fb639 100644 --- a/subworkflows/local/peaks.nf +++ b/subworkflows/local/peaks.nf @@ -39,8 +39,8 @@ workflow CALL_PEAKS { Channel.fromPath("${params.genomes[ params.genome ].chromosomes_dir}", type: 'dir', checkIfExists: true) .set{ chrom_files } - ch_tagalign | MACS_BROAD - ch_tagalign | MACS_NARROW + ch_tagalign.combine(effective_genome_size) | MACS_BROAD + ch_tagalign.combine(effective_genome_size) | MACS_NARROW ch_tagalign | SICER GEM(ch_gem, chrom_files) From bf5bca182a8401e05682d407da19011576113912 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 2 Oct 2023 16:52:29 -0400 Subject: [PATCH 028/100] fix: typo --- modules/local/peaks.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/peaks.nf b/modules/local/peaks.nf index 27cbe0af..53d9fb12 100644 --- a/modules/local/peaks.nf +++ b/modules/local/peaks.nf @@ -13,7 +13,7 @@ process CALC_GENOME_FRAC { script: """ - genome_frac=`calc_effective_genome_fraction.py effective_genome_size ${chrom_sizes} ${params.deeptools.excluded_chroms}` + genome_frac=`calc_effective_genome_fraction.py ${effective_genome_size} ${chrom_sizes} ${params.deeptools.excluded_chroms}` echo \$genome_frac """ From 62c2b5a4ea4b4a35dc1226b994cfa8fdda19f077 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 2 Oct 2023 16:52:54 -0400 Subject: [PATCH 029/100] fix: path to mqc config --- nextflow.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index 7580f0e8..10645258 100644 --- a/nextflow.config +++ b/nextflow.config @@ -38,7 +38,7 @@ params { normalize_using = "RPGC" excluded_chroms = "chrM chrX chrY" } - multiqc_config = "${baseDir}assets/multiqc_config.yaml" + multiqc_config = "${baseDir}/assets/multiqc_config.yaml" min_fragment_length = 200 // https://github.com/CCBR/Pipeliner/blob/86c6ccaa3d58381a0ffd696bbf9c047e4f991f9e/Rules/InitialChIPseqQC.snakefile#L539 gem_read_dists = 'https://groups.csail.mit.edu/cgs/gem/download/Read_Distribution_default.txt' From a891a64693fcf2d3861a331ec8d2bdb091595dc1 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 2 Oct 2023 16:53:37 -0400 Subject: [PATCH 030/100] refactor: get chrom_files channel in PREPARE_GENOME subwf --- main.nf | 6 ++++-- subworkflows/local/peaks.nf | 3 +-- subworkflows/local/prepare_genome.nf | 3 +++ 3 files changed, 8 insertions(+), 4 deletions(-) diff --git a/main.nf b/main.nf index 3c6456f4..2e2ae991 100644 --- a/main.nf +++ b/main.nf @@ -40,12 +40,14 @@ workflow { TRIM_SE.out.set{ trimmed_fastqs } PREPARE_GENOME() + chrom_sizes = PREPARE_GENOME.out.chrom_sizes + effective_genome_size = PREPARE_GENOME.out.effective_genome_size ALIGN_BLACKLIST(trimmed_fastqs, PREPARE_GENOME.out.blacklist_files, PREPARE_GENOME.out.blacklist_name) ALIGN_GENOME(ALIGN_BLACKLIST.out.reads, PREPARE_GENOME.out.reference_files) ALIGN_GENOME.out.bam.set{ aligned_bam } - DEDUPLICATE(aligned_bam.combine(PREPARE_GENOME.out.chrom_sizes), effective_genome_size) + DEDUPLICATE(aligned_bam.combine(chrom_sizes), effective_genome_size) DEDUPLICATE.out.bam.set{ deduped_bam } DEDUPLICATE.out.tag_align.set{ deduped_tagalign } @@ -69,6 +71,6 @@ workflow { } if (params.run.call_peaks) { - CALL_PEAKS(chrom_sizes, deduped_tagalign, frag_lengths, effective_genome_size) + CALL_PEAKS(chrom_sizes, PREPARE_GENOME.out.chrom_files, deduped_tagalign, frag_lengths, effective_genome_size) } } diff --git a/subworkflows/local/peaks.nf b/subworkflows/local/peaks.nf index 739fb639..61427b38 100644 --- a/subworkflows/local/peaks.nf +++ b/subworkflows/local/peaks.nf @@ -9,6 +9,7 @@ include { GEM } from "../../modules/local/peaks.nf" workflow CALL_PEAKS { take: chrom_sizes + chrom_files deduped_tagalign frag_lengths effective_genome_size @@ -36,8 +37,6 @@ workflow CALL_PEAKS { .combine(Channel.fromPath(params.gem_read_dists, checkIfExists: true)) .combine(chrom_sizes) .set { ch_gem } - Channel.fromPath("${params.genomes[ params.genome ].chromosomes_dir}", type: 'dir', checkIfExists: true) - .set{ chrom_files } ch_tagalign.combine(effective_genome_size) | MACS_BROAD ch_tagalign.combine(effective_genome_size) | MACS_NARROW diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index e077007d..d6e16a0a 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -14,6 +14,8 @@ workflow PREPARE_GENOME { .set{ ch_reference_files } Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) .set{ ch_chrom_sizes } + Channel.fromPath("${params.genomes[ params.genome ].chromosomes_dir}", type: 'dir', checkIfExists: true) + .set{ ch_chrom_files } Channel.fromPath(params.genomes[ params.genome ].gene_info, checkIfExists: true) .set{ ch_gene_info } @@ -26,6 +28,7 @@ workflow PREPARE_GENOME { blacklist_name = ch_blacklist_name reference_files = ch_reference_files chrom_sizes = ch_chrom_sizes + chrom_files = ch_chrom_files gene_info = ch_gene_info effective_genome_size = ch_egs } From 9c56dd9f6600bbb199f95f7f74df2e14037bd4f9 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 2 Oct 2023 19:04:24 -0400 Subject: [PATCH 031/100] feat: use bwa/index & khmer/uniquekmers from nf-core --- conf/test.config | 3 ++ conf/test_mm10.config | 1 + conf/test_paired.config | 2 + modules.json | 33 ++++++++++++++ modules/CCBR/bwa/mem/main.nf | 38 ++++++++++++++++ modules/CCBR/bwa/mem/meta.yml | 47 +++++++++++++++++++ modules/local/align.nf | 2 +- modules/nf-core/bwa/index/main.nf | 53 ++++++++++++++++++++++ modules/nf-core/bwa/index/meta.yml | 42 +++++++++++++++++ modules/nf-core/khmer/uniquekmers/main.nf | 38 ++++++++++++++++ modules/nf-core/khmer/uniquekmers/meta.yml | 42 +++++++++++++++++ nextflow.config | 1 + subworkflows/local/prepare_genome.nf | 7 +++ 13 files changed, 308 insertions(+), 1 deletion(-) create mode 100644 modules.json create mode 100644 modules/CCBR/bwa/mem/main.nf create mode 100644 modules/CCBR/bwa/mem/meta.yml create mode 100644 modules/nf-core/bwa/index/main.nf create mode 100644 modules/nf-core/bwa/index/meta.yml create mode 100644 modules/nf-core/khmer/uniquekmers/main.nf create mode 100644 modules/nf-core/khmer/uniquekmers/meta.yml diff --git a/conf/test.config b/conf/test.config index 9df0a609..204ec63a 100644 --- a/conf/test.config +++ b/conf/test.config @@ -4,6 +4,9 @@ params { outdir = 'results/test' input = 'assets/samplesheet_test.csv' // adapted from https://github.com/nf-core/test-datasets/blob/chipseq/samplesheet/v2.0/samplesheet_test.csv + + read_length = 50 + genome = 'hg38' max_cpus = 32 // for interactive node on biowulf diff --git a/conf/test_mm10.config b/conf/test_mm10.config index 8e2a3963..0b40733e 100644 --- a/conf/test_mm10.config +++ b/conf/test_mm10.config @@ -5,4 +5,5 @@ params { genome = 'mm10' outdir = "results/test_mm10" input = "assets/samplesheet_test_mm10.csv" + read_length = 50 } diff --git a/conf/test_paired.config b/conf/test_paired.config index 8c4c2dee..e2cb364f 100644 --- a/conf/test_paired.config +++ b/conf/test_paired.config @@ -4,4 +4,6 @@ params { outdir = 'results/test_paired' input = 'https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/samplesheet/v2.0/samplesheet_test.csv' + + read_length = 50 } diff --git a/modules.json b/modules.json new file mode 100644 index 00000000..e4b4b417 --- /dev/null +++ b/modules.json @@ -0,0 +1,33 @@ +{ + "name": "CCBR/CHAMPAGNE", + "homePage": "https://github.com/CCBR/CHAMPAGNE", + "repos": { + "https://github.com/CCBR/nf-modules": { + "modules": { + "CCBR": { + "bwa/mem": { + "branch": "main", + "git_sha": "a4b80fe5424b4bef8ebb6c35c2e7f3c0a9caaa81", + "installed_by": ["modules"] + } + } + } + }, + "https://github.com/nf-core/modules.git": { + "modules": { + "nf-core": { + "bwa/index": { + "branch": "master", + "git_sha": "bfed129da5134b4439b1821c917972570d44d39c", + "installed_by": ["modules"] + }, + "khmer/uniquekmers": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"] + } + } + } + } + } +} diff --git a/modules/CCBR/bwa/mem/main.nf b/modules/CCBR/bwa/mem/main.nf new file mode 100644 index 00000000..65bb3a61 --- /dev/null +++ b/modules/CCBR/bwa/mem/main.nf @@ -0,0 +1,38 @@ +process BWA_MEM { + tag { meta.id } + label 'process_higher' + + container = 'nciccbr/ccbr_ubuntu_base_20.04:v5' + + input: + tuple val(meta), path(fastq) + path(index_files) + val(index_name) + + output: + tuple val(meta), path("*.bam"), emit: bam + path "versions.yml" , emit: versions + + script: + def prefix = task.ext.prefix ?: "${meta.id}" + """ + # current working directory is a tmpdir when 'scratch' is set + TMP=tmp/ + mkdir \$TMP + trap 'rm -rf "\$TMP"' EXIT + + bwa mem -t ${task.cpus} ${index_name} ${fastq} -o \$TMP/align.bam + + samtools sort \\ + -@ ${task.cpus} \\ + -m 2G \\ + -T \$TMP \\ + \$TMP/align.bam > ${prefix}.bam + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bwa: \$(echo \$(bwa 2>&1) | sed 's/^.*Version: //; s/Contact:.*\$//') + samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') + END_VERSIONS + """ +} diff --git a/modules/CCBR/bwa/mem/meta.yml b/modules/CCBR/bwa/mem/meta.yml new file mode 100644 index 00000000..3f8677ef --- /dev/null +++ b/modules/CCBR/bwa/mem/meta.yml @@ -0,0 +1,47 @@ +name: bwa_mem +description: Performs fastq alignment to a fasta reference using BWA. Adapted from the nf-core bwa-mem module. +keywords: + - mem + - bwa + - alignment + - map + - fastq + - bam + - sam +tools: + - bwa: + description: | + BWA is a software package for mapping DNA sequences against + a large reference genome, such as the human genome. + homepage: http://bio-bwa.sourceforge.net/ + documentation: http://www.htslib.org/doc/samtools.html + arxiv: arXiv:1303.3997 + licence: ["GPL-3.0-or-later"] +input: + - meta: + type: map + description: | + Groovy Map containing sample information + e.g. [ id:'test', single_end:false ] + - reads: + type: file + description: | + List of input FastQ files of size 1 and 2 for single-end and paired-end data, + respectively. + - index: + type: file + description: BWA genome index files + pattern: "Directory containing BWA index *.{amb,ann,bwt,pac,sa}" +output: + - bam: + type: file + description: Output BAM file containing read alignments + pattern: "*.{bam}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@drpatelh" + - "@jeremy1805" + - "@kelly-sovacool" diff --git a/modules/local/align.nf b/modules/local/align.nf index ff5e4dbb..b2d0a30d 100644 --- a/modules/local/align.nf +++ b/modules/local/align.nf @@ -1,5 +1,5 @@ -process ALIGN_BLACKLIST { +process ALIGN_BLACKLIST { // TODO: refactor this as a subworkflow that combines alignment and filtering processes tag { meta.id } label 'align' label 'process_higher' diff --git a/modules/nf-core/bwa/index/main.nf b/modules/nf-core/bwa/index/main.nf new file mode 100644 index 00000000..c30d194d --- /dev/null +++ b/modules/nf-core/bwa/index/main.nf @@ -0,0 +1,53 @@ +process BWA_INDEX { + tag "$fasta" + label 'process_single' + + conda "bioconda::bwa=0.7.17" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/bwa:0.7.17--hed695b0_7' : + 'biocontainers/bwa:0.7.17--hed695b0_7' }" + + input: + tuple val(meta), path(fasta) + + output: + tuple val(meta), path(bwa) , emit: index + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def prefix = task.ext.prefix ?: "${fasta.baseName}" + def args = task.ext.args ?: '' + """ + mkdir bwa + bwa \\ + index \\ + $args \\ + -p bwa/${prefix} \\ + $fasta + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bwa: \$(echo \$(bwa 2>&1) | sed 's/^.*Version: //; s/Contact:.*\$//') + END_VERSIONS + """ + + stub: + def prefix = task.ext.prefix ?: "${fasta.baseName}" + """ + mkdir bwa + + touch bwa/${prefix}.amb + touch bwa/${prefix}.ann + touch bwa/${prefix}.bwt + touch bwa/${prefix}.pac + touch bwa/${prefix}.sa + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bwa: \$(echo \$(bwa 2>&1) | sed 's/^.*Version: //; s/Contact:.*\$//') + END_VERSIONS + """ +} diff --git a/modules/nf-core/bwa/index/meta.yml b/modules/nf-core/bwa/index/meta.yml new file mode 100644 index 00000000..2c6cfcd7 --- /dev/null +++ b/modules/nf-core/bwa/index/meta.yml @@ -0,0 +1,42 @@ +name: bwa_index +description: Create BWA index for reference genome +keywords: + - index + - fasta + - genome + - reference +tools: + - bwa: + description: | + BWA is a software package for mapping DNA sequences against + a large reference genome, such as the human genome. + homepage: http://bio-bwa.sourceforge.net/ + documentation: http://www.htslib.org/doc/samtools.html + arxiv: arXiv:1303.3997 + licence: ["GPL-3.0-or-later"] +input: + - meta: + type: map + description: | + Groovy Map containing reference information. + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: Input genome fasta file +output: + - meta: + type: map + description: | + Groovy Map containing reference information. + e.g. [ id:'test', single_end:false ] + - index: + type: file + description: BWA genome index files + pattern: "*.{amb,ann,bwt,pac,sa}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@drpatelh" + - "@maxulysse" diff --git a/modules/nf-core/khmer/uniquekmers/main.nf b/modules/nf-core/khmer/uniquekmers/main.nf new file mode 100644 index 00000000..9576034f --- /dev/null +++ b/modules/nf-core/khmer/uniquekmers/main.nf @@ -0,0 +1,38 @@ +process KHMER_UNIQUEKMERS { + tag "$fasta" + label 'process_low' + + conda "bioconda::khmer=3.0.0a3" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/khmer:3.0.0a3--py37haa7609a_2' : + 'biocontainers/khmer:3.0.0a3--py37haa7609a_2' }" + + input: + path fasta + val kmer_size + + output: + path "report.txt" , emit: report + path "kmers.txt" , emit: kmers + path "versions.yml", emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + """ + unique-kmers.py \\ + -k $kmer_size \\ + -R report.txt \\ + $args \\ + $fasta + + grep ^number report.txt | sed 's/^.*:.[[:blank:]]//g' > kmers.txt + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + khmer: \$( unique-kmers.py --version 2>&1 | grep ^khmer | sed 's/^khmer //;s/ .*\$//' ) + END_VERSIONS + """ +} diff --git a/modules/nf-core/khmer/uniquekmers/meta.yml b/modules/nf-core/khmer/uniquekmers/meta.yml new file mode 100644 index 00000000..f9f63972 --- /dev/null +++ b/modules/nf-core/khmer/uniquekmers/meta.yml @@ -0,0 +1,42 @@ +name: "khmer_uniquekmers" +description: In-memory nucleotide sequence k-mer counting, filtering, graph traversal and more +keywords: + - khmer + - k-mer + - effective genome size + +tools: + - "khmer": + description: khmer k-mer counting library + homepage: https://github.com/dib-lab/khmer + documentation: https://khmer.readthedocs.io/en/latest/ + tool_dev_url: https://github.com/dib-lab/khmer + doi: "10.12688/f1000research.6924.1" + licence: ["BSD License"] + +input: + - fasta: + type: file + description: fasta file + pattern: "*.{fa,fasta}" + - kmer_size: + type: value + description: k-mer size to use + pattern: "[0-9]+" + +output: + - report: + type: file + description: Text file containing unique-kmers.py execution report + pattern: "report.txt" + - kmers: + type: file + description: Text file containing number of kmers + pattern: "kmers.txt" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" + +authors: + - "@JoseEspinosa" diff --git a/nextflow.config b/nextflow.config index 10645258..210b172b 100644 --- a/nextflow.config +++ b/nextflow.config @@ -3,6 +3,7 @@ nextflow.enable.dsl = 2 params { input = null seq_center = null + read_length = null genome = null fasta = null gtf = null diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index d6e16a0a..fa40dfb9 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -1,3 +1,6 @@ +include { BWA_INDEX } from "../../modules/nf-core/bwa/index" +include { KHMER_UNIQUEKMERS } from '../../modules/nf-core/khmer/uniquekmers' + workflow PREPARE_GENOME { main: ch_blacklist_name = params.genomes[ params.genome ].blacklist ? Channel.value(params.genomes[ params.genome ].blacklist) : "${params.genome}.blacklist" @@ -5,6 +8,10 @@ workflow PREPARE_GENOME { if (params.fasta && params.gtf) { ch_fasta = file(params.fasta) ch_gtf = file(params.gtf) + + BWA_INDEX(ch_fasta) + KHMER_UNIQUEKMERS(ch_fasta, params.read_length) + } else { Channel.fromPath(params.genomes[ params.genome ].blacklist_files, checkIfExists: true) .collect() From 693acf0013fa21e8f167864b41a8039bdd2a0542 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 14:03:39 -0400 Subject: [PATCH 032/100] feat: use new picard container has pigz & better picard api resolves #106 --- conf/containers.config | 2 +- modules/local/align.nf | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/conf/containers.config b/conf/containers.config index b334600d..914ddfbd 100644 --- a/conf/containers.config +++ b/conf/containers.config @@ -11,7 +11,7 @@ params { multiqc = 'nciccbr/ccbr_multiqc_1.15:v1' ngsqc = 'nciccbr/ccbr_ngsqc_0.31:v1' phantom_peaks = 'quay.io/biocontainers/phantompeakqualtools:1.2.2--hdfd78af_1' - picard = 'quay.io/biocontainers/picard:2.27.4--hdfd78af_0' + picard = 'nciccbr/ccbr_picard_2.27.5:v1' preseq = 'nciccbr/ccbr_preseq_v2.0:v1' r = 'nciccbr/ccbr_r_4.3.0:v1' sicer = 'nciccbr/ccbr_sicer2_1.0.3:v1' diff --git a/modules/local/align.nf b/modules/local/align.nf index 3241347c..48c6cbac 100644 --- a/modules/local/align.nf +++ b/modules/local/align.nf @@ -55,7 +55,7 @@ process BAM_TO_FASTQ { --FASTQ ${bam.baseName}.R1.fastq \\ --SECOND_END_FASTQ ${bam.baseName}.R2.fastq \\ --UNPAIRED_FASTQ ${bam.baseName}.unpaired.fastq - # TODO: gzip output. need container w/ both picard & pigz + pigz -p ${task.cpus} *.fastq """ } stub: From 307fcd7fe38bbef5b5888d39754634d75f5f8ff4 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 14:07:19 -0400 Subject: [PATCH 033/100] chore: update ccbr nf-modules --- modules.json | 10 +++++-- modules/CCBR/bwa/index/main.nf | 47 +++++++++++++++++++++++++++++++++ modules/CCBR/bwa/index/meta.yml | 41 ++++++++++++++++++++++++++++ modules/CCBR/bwa/mem/main.nf | 12 ++++++--- modules/CCBR/bwa/mem/meta.yml | 2 -- 5 files changed, 104 insertions(+), 8 deletions(-) create mode 100644 modules/CCBR/bwa/index/main.nf create mode 100644 modules/CCBR/bwa/index/meta.yml diff --git a/modules.json b/modules.json index bfc19361..cbdfea8a 100644 --- a/modules.json +++ b/modules.json @@ -5,9 +5,14 @@ "https://github.com/CCBR/nf-modules": { "modules": { "CCBR": { + "bwa/index": { + "branch": "main", + "git_sha": "b39dceb9d9dd55804f8cd81181a315a48dc0f4c6", + "installed_by": ["modules"] + }, "bwa/mem": { "branch": "main", - "git_sha": "a4b80fe5424b4bef8ebb6c35c2e7f3c0a9caaa81", + "git_sha": "520860abefe5e45ac604e92051d2daebdae4aa81", "installed_by": ["modules"] }, "cutadapt": { @@ -28,7 +33,8 @@ }, "khmer/uniquekmers": { "branch": "master", - "git_sha": "911696ea0b62df80e900ef244d7867d177971f73" + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"] } } } diff --git a/modules/CCBR/bwa/index/main.nf b/modules/CCBR/bwa/index/main.nf new file mode 100644 index 00000000..a30f3d76 --- /dev/null +++ b/modules/CCBR/bwa/index/main.nf @@ -0,0 +1,47 @@ +process BWA_INDEX { + tag { fasta } + label 'process_single' + + container 'nciccbr/ccbr_ubuntu_base_20.04:v5' + + input: + tuple val(meta), path(fasta) + + output: + tuple val(meta), path(bwa) , emit: index + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def prefix = task.ext.prefix ?: "${fasta.baseName}" + def args = task.ext.args ?: '' + """ + mkdir bwa + bwa \\ + index \\ + ${args} \\ + -p bwa/${prefix} \\ + ${fasta} + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bwa: \$(echo \$(bwa 2>&1) | sed 's/^.*Version: //; s/Contact:.*\$//') + END_VERSIONS + """ + + stub: + def prefix = task.ext.prefix ?: "${fasta.baseName}" + """ + mkdir bwa + for ext in amb ann bwt pac sa; do + touch bwa/${prefix}.\$ext + done + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bwa: \$(echo \$(bwa 2>&1) | sed 's/^.*Version: //; s/Contact:.*\$//') + END_VERSIONS + """ +} diff --git a/modules/CCBR/bwa/index/meta.yml b/modules/CCBR/bwa/index/meta.yml new file mode 100644 index 00000000..02a14128 --- /dev/null +++ b/modules/CCBR/bwa/index/meta.yml @@ -0,0 +1,41 @@ +name: bwa_index +description: Create BWA index for reference genome. Adapted from the nf-core bwa index module. +keywords: + - index + - fasta + - genome + - reference +tools: + - bwa: + description: | + BWA is a software package for mapping DNA sequences against + a large reference genome, such as the human genome. + homepage: http://bio-bwa.sourceforge.net/ + documentation: http://www.htslib.org/doc/samtools.html + arxiv: arXiv:1303.3997 + licence: ["GPL-3.0-or-later"] +input: + - meta: + type: map + description: | + Groovy Map containing reference information. + e.g. [ id:'test', single_end:false ] + - fasta: + type: file + description: Input genome fasta file +output: + - meta: + type: map + description: | + Groovy Map containing reference information. + e.g. [ id:'test', single_end:false ] + - index: + type: file + description: BWA genome index files + pattern: "*.{amb,ann,bwt,pac,sa}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@kelly-sovacool" diff --git a/modules/CCBR/bwa/mem/main.nf b/modules/CCBR/bwa/mem/main.nf index 65bb3a61..4a8fd260 100644 --- a/modules/CCBR/bwa/mem/main.nf +++ b/modules/CCBR/bwa/mem/main.nf @@ -2,17 +2,19 @@ process BWA_MEM { tag { meta.id } label 'process_higher' - container = 'nciccbr/ccbr_ubuntu_base_20.04:v5' + container 'nciccbr/ccbr_ubuntu_base_20.04:v5' input: tuple val(meta), path(fastq) - path(index_files) - val(index_name) + tuple val(meta_idx), path(index_files) output: tuple val(meta), path("*.bam"), emit: bam path "versions.yml" , emit: versions + when: + task.ext.when == null || task.ext.when + script: def prefix = task.ext.prefix ?: "${meta.id}" """ @@ -21,7 +23,9 @@ process BWA_MEM { mkdir \$TMP trap 'rm -rf "\$TMP"' EXIT - bwa mem -t ${task.cpus} ${index_name} ${fastq} -o \$TMP/align.bam + INDEX=`find -L ./ -name "*.amb" | sed 's/\\.amb\$//'` + + bwa mem -t ${task.cpus} \$INDEX ${fastq} -o \$TMP/align.bam samtools sort \\ -@ ${task.cpus} \\ diff --git a/modules/CCBR/bwa/mem/meta.yml b/modules/CCBR/bwa/mem/meta.yml index 3f8677ef..14feefe9 100644 --- a/modules/CCBR/bwa/mem/meta.yml +++ b/modules/CCBR/bwa/mem/meta.yml @@ -42,6 +42,4 @@ output: description: File containing software versions pattern: "versions.yml" authors: - - "@drpatelh" - - "@jeremy1805" - "@kelly-sovacool" From 6226912a54ad13ee2f9901fe730719a7a3f313a6 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 14:08:16 -0400 Subject: [PATCH 034/100] chore: mark CCBR modules as generated --- .gitattributes | 2 ++ 1 file changed, 2 insertions(+) diff --git a/.gitattributes b/.gitattributes index 0b9f4afd..096d3cf6 100644 --- a/.gitattributes +++ b/.gitattributes @@ -2,6 +2,8 @@ *.nf.test linguist-language=nextflow modules/nf-core/** linguist-generated subworkflows/nf-core/** linguist-generated +modules/CCBR/** linguist-generated +subworkflows/CCBR/** linguist-generated *.config linguist-language=nextflow assets/** linguist-generated docs/**.html linguist-generated From 54cb8b9a3bc6ea0de7b4bf5840cc9d174e53f167 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 14:09:05 -0400 Subject: [PATCH 035/100] chore: assets aren't necessarily generated --- .gitattributes | 1 - 1 file changed, 1 deletion(-) diff --git a/.gitattributes b/.gitattributes index 096d3cf6..daa4fa2d 100644 --- a/.gitattributes +++ b/.gitattributes @@ -5,5 +5,4 @@ subworkflows/nf-core/** linguist-generated modules/CCBR/** linguist-generated subworkflows/CCBR/** linguist-generated *.config linguist-language=nextflow -assets/** linguist-generated docs/**.html linguist-generated From ab9fd7e8c2adeb4dac9ca443b2fb4c96f86e1fe4 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 14:10:24 -0400 Subject: [PATCH 036/100] chore: remove duplicate line in .gitattributes --- .gitattributes | 1 - 1 file changed, 1 deletion(-) diff --git a/.gitattributes b/.gitattributes index daa4fa2d..547ee638 100644 --- a/.gitattributes +++ b/.gitattributes @@ -4,5 +4,4 @@ modules/nf-core/** linguist-generated subworkflows/nf-core/** linguist-generated modules/CCBR/** linguist-generated subworkflows/CCBR/** linguist-generated -*.config linguist-language=nextflow docs/**.html linguist-generated From 69e5f66fab747966685bd75813157e7651d5acd7 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 14:39:55 -0400 Subject: [PATCH 037/100] fix: use effective genome size from prep_genome subwf --- main.nf | 2 +- modules/local/peaks.nf | 4 ++-- subworkflows/local/peaks.nf | 5 +++-- 3 files changed, 6 insertions(+), 5 deletions(-) diff --git a/main.nf b/main.nf index c5f135c4..8ff04754 100644 --- a/main.nf +++ b/main.nf @@ -78,7 +78,7 @@ workflow { } if (params.run.call_peaks) { - CALL_PEAKS(chrom_sizes, PREPARE_GENOME.out.chrom_files, deduped_tagalign, frag_lengths, effective_genome_size) + CALL_PEAKS(chrom_sizes, PREPARE_GENOME.out.chrom_files, deduped_tagalign, deduped_bam, frag_lengths, effective_genome_size) ch_multiqc = ch_multiqc.mix(CALL_PEAKS.out.plots) } diff --git a/modules/local/peaks.nf b/modules/local/peaks.nf index 245d07b7..334937e9 100644 --- a/modules/local/peaks.nf +++ b/modules/local/peaks.nf @@ -218,7 +218,7 @@ process GEM { container = "${params.containers.gem}" input: - tuple val(meta), path(chip), path(input), val(format), path(read_dists), path(chrom_sizes), path(chrom_dir) + tuple val(meta), path(chip), path(input), val(format), path(read_dists), path(chrom_sizes), path(chrom_dir), val(effective_genome_size) output: tuple val(meta), path("${meta.id}/*GEM_events.narrowPeak"), val("${task.process.tokenize(':')[-1].toLowerCase()}"), emit: peak @@ -233,7 +233,7 @@ process GEM { --d ${read_dists} \\ --g ${chrom_sizes} \\ --genome ${chrom_dir} \\ - --s ${params.genomes[ params.genome ].effective_genome_size} \\ + --s ${effective_genome_size} \\ --expt ${chip} \\ --ctrl ${input} \\ --f ${f} \\ diff --git a/subworkflows/local/peaks.nf b/subworkflows/local/peaks.nf index fb49bccf..434f7a27 100644 --- a/subworkflows/local/peaks.nf +++ b/subworkflows/local/peaks.nf @@ -74,13 +74,14 @@ workflow CALL_PEAKS { ch_tagalign .combine(Channel.fromPath(params.gem.read_dists, checkIfExists: true)) .combine(chrom_sizes) - .combine(Channel.fromPath("${params.genomes[ params.genome ].chromosomes_dir}", type: 'dir', checkIfExists: true)) + .combine(chrom_files) + .combine(effective_genome_size) .set { ch_gem } ch_macs | MACS_BROAD ch_macs | MACS_NARROW ch_sicer | SICER | CONVERT_SICER - GEM(ch_gem, chrom_files) + GEM(ch_gem) CONVERT_SICER.out.peak .mix(GEM.out.peak, From e3c658e6294c11a2b9c8fc6f4b3d8312db4f3cea Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 23:05:26 -0400 Subject: [PATCH 038/100] feat: copy splitRef.py from biowulf /data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/Chromsomes/splitRef.py --- bin/splitRef.py | 59 +++++++++++++++++++++++++++++++++++++++++++++++++ 1 file changed, 59 insertions(+) create mode 100644 bin/splitRef.py diff --git a/bin/splitRef.py b/bin/splitRef.py new file mode 100644 index 00000000..6cb32fb0 --- /dev/null +++ b/bin/splitRef.py @@ -0,0 +1,59 @@ +#!/usr/bin/env python + +from __future__ import print_function +import sys + + +def formatSequencelength(seq, stringlen): + fseq = "" + for i in range(len(seq)): + index = i + 1 + if index % 80 == 0: + fseq += "{}{}".format(seq[i], "\n") + else: + fseq += seq[i] + return fseq + + +def parsed(filename): + fh = open(filename, "r") + sequence = "" + chrom = "" + seqindex = 0 + seqlen = 0 + for line in fh: + line = line.strip() + if line.startswith(">") and sequence != "": + yield chrom, formatSequencelength(sequence, seqlen), len(sequence) + chrom = line.split(" ")[0] + sequence = "" + elif line.startswith(">"): + chrom = line.split(" ")[0] + else: + seqindex += 1 + sequence += line + if seqindex == 1: + seqlen = len(line) + else: + # formatSequencelength(sequence, seqlen) + yield chrom, formatSequencelength(sequence, seqlen), len(sequence) + fh.close() + + +def main(): + file = sys.argv[1] + chromsizesfh = open("chrom.sizes", "w") + + for chrom, seq, chromsize in parsed(file): + chromsizesfh.write("{}\t{}\n".format(chrom.replace(">", ""), chromsize)) + outfilename = chrom.replace(">", "") + ".fa" + outfh = open(outfilename, "w") + print("{}\n".format(chrom)) + outfh.write("{}\n{}\n".format(chrom, seq.rstrip())) + outfh.close() + + chromsizesfh.close() + + +if __name__ == "__main__": + main() From 2d7dd9c811079acb355a18a91da7b012c4fb805b Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 23:16:16 -0400 Subject: [PATCH 039/100] feat: use better blacklists w/ MT & Ribosomal seqs --- conf/genomes.config | 10 ++++------ 1 file changed, 4 insertions(+), 6 deletions(-) diff --git a/conf/genomes.config b/conf/genomes.config index 376054e0..e75b2404 100644 --- a/conf/genomes.config +++ b/conf/genomes.config @@ -1,18 +1,16 @@ params { genomes { 'hg38' { - blacklist = 'hg38.blacklist' - blacklist_files = "${params.index_dir}/hg38_basic/indexes/hg38.blacklist*" - reference_files = "${params.index_dir}/hg38_basic/indexes/hg38*" + blacklist_index = "${params.index_dir}/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.*" + reference_index = "${params.index_dir}/hg38_basic/indexes/hg38*" effective_genome_size = 2700000000 chrom_sizes = "${params.index_dir}/hg38_basic/indexes/hg38.fa.sizes" gene_info = "${params.index_dir}/hg38_basic/geneinfo.bed" chromosomes_dir = "${params.index_dir}/hg38_basic/Chromsomes/" } 'mm10' { - blacklist = 'mm10.blacklist' - blacklist_files = "${params.index_dir}/mm10_basic/indexes/mm10.blacklist*" - reference_files = "${params.index_dir}/mm10_basic/indexes/mm10*" + blacklist_index = "${params.index_dir}/mm10_basic/indexes/mm10.blacklist.chrM.chr_rDNA.*" + reference_index = "${params.index_dir}/mm10_basic/indexes/mm10*" effective_genome_size = 2400000000 chrom_sizes = "${params.index_dir}/mm10_basic/indexes/mm10.fa.sizes" gene_info = "${params.index_dir}/mm10_basic/geneinfo.bed" From 9523c2911c70e51e1d07d46b961e08e5a50561fa Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 23:16:56 -0400 Subject: [PATCH 040/100] refactor: splitRef.py to accept CLI args for file names --- bin/splitRef.py | 12 ++++++------ 1 file changed, 6 insertions(+), 6 deletions(-) diff --git a/bin/splitRef.py b/bin/splitRef.py index 6cb32fb0..b5d9bed4 100644 --- a/bin/splitRef.py +++ b/bin/splitRef.py @@ -1,6 +1,7 @@ #!/usr/bin/env python from __future__ import print_function +import os import sys @@ -40,13 +41,12 @@ def parsed(filename): fh.close() -def main(): - file = sys.argv[1] - chromsizesfh = open("chrom.sizes", "w") +def main(fasta_fn, chrom_sizes_fn, outdir): + chromsizesfh = open(chrom_sizes_fn, "w") - for chrom, seq, chromsize in parsed(file): + for chrom, seq, chromsize in parsed(fasta_fn): chromsizesfh.write("{}\t{}\n".format(chrom.replace(">", ""), chromsize)) - outfilename = chrom.replace(">", "") + ".fa" + outfilename = os.path.join(outdir, chrom.replace(">", "") + ".fa") outfh = open(outfilename, "w") print("{}\n".format(chrom)) outfh.write("{}\n{}\n".format(chrom, seq.rstrip())) @@ -56,4 +56,4 @@ def main(): if __name__ == "__main__": - main() + main(sys.argv[1], sys.argv[2], sys.argv[3]) From eb821e224507aecd41e984a525622ba02c4164a3 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 23:18:27 -0400 Subject: [PATCH 041/100] feat: custom split_ref_chroms process and gtf2bed --- modules/local/prepare_genome.nf | 42 +++++++++++++++++++++++++++++++++ 1 file changed, 42 insertions(+) diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index e69de29b..a9e37a8b 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -0,0 +1,42 @@ +process GTF2BED { + tag { gtf } + label 'process_single' + + container 'quay.io/biocontainers/bedops:2.4.41--h4ac6f70_1' + + input: + path(gtf) + + output: + path("${gtf.baseName}.bed") + + script: + """ + cat ${gtf} | gtf2bed > ${gtf.baseName}.bed + """ + stub: + """ + touch ${gtf.baseName}.bed + """ +} +process SPLIT_REF_CHROMS { + tag { fasta } + label 'process_single' + container "${params.containers.base}" + + input: + path(fasta) + + output: + path("${fasta.baseName}.chrom.sizes"), emit: chrom_sizes + path("chroms/*") , emit: chrom_files + + script: + """ + splitRef.py ${fasta} ${fasta.baseName}.chrom.sizes chroms + """ + stub: + """ + touch ${fasta.baseName}.chrom.sizes + """ +} From ca58d49de7ab700f4aa72030645f1f2fad4198a9 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 23:18:50 -0400 Subject: [PATCH 042/100] test: use custom genome --- conf/test.config | 9 +++++++-- 1 file changed, 7 insertions(+), 2 deletions(-) diff --git a/conf/test.config b/conf/test.config index 65bcb0da..67ea9203 100644 --- a/conf/test.config +++ b/conf/test.config @@ -7,7 +7,12 @@ params { read_length = 50 - genome = 'hg38' + // Genome references + genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' + genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' + blacklist = '' + genome = 'custom_test' + save_reference = true // https://github.com/nf-core/chipseq/blob/51eba00b32885c4d0bec60db3cb0a45eb61e34c5/subworkflows/local/prepare_genome.nf#L47-L51 max_cpus = 32 // for interactive node on biowulf max_memory = '120 GB' // for interactive node on biowulf @@ -16,7 +21,7 @@ params { run { qc = false normalize_input = false - call_peaks = false // no peaks exist with this yeast dataset with hg38 as the reference + call_peaks = false } } dag { From aec8aea0ee373369a98e979c73139b63bbba0df9 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 23:19:23 -0400 Subject: [PATCH 043/100] refactor: use custom genome if genome fasta & gtf files given, otherwise use built-in --- subworkflows/local/prepare_genome.nf | 45 ++++++++++++++++++---------- 1 file changed, 29 insertions(+), 16 deletions(-) diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index fa40dfb9..ff08e5ec 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -1,24 +1,38 @@ -include { BWA_INDEX } from "../../modules/nf-core/bwa/index" -include { KHMER_UNIQUEKMERS } from '../../modules/nf-core/khmer/uniquekmers' +include { BWA_INDEX as BWA_INDEX_BL + BWA_INDEX as BWA_INDEX_REF } from "../../modules/CCBR/bwa/index" +include { KHMER_UNIQUEKMERS } from '../../modules/nf-core/khmer/uniquekmers' +include { SPLIT_REF_CHROMS } from '../../modules/local/prepare_genome.nf' +include { GTF2BED } from '../../modules/local/prepare_genome.nf' workflow PREPARE_GENOME { main: - ch_blacklist_name = params.genomes[ params.genome ].blacklist ? Channel.value(params.genomes[ params.genome ].blacklist) : "${params.genome}.blacklist" - - if (params.fasta && params.gtf) { - ch_fasta = file(params.fasta) - ch_gtf = file(params.gtf) - - BWA_INDEX(ch_fasta) + if (params.genome_fasta && params.genes_gtf) { + ch_fasta = file(params.genome_fasta) + ch_blacklist_input = file(params.blacklist) + if (ch_blacklist_input.endsWith('.fa') || ch_blacklist_input.endsWith('.fna') || ch_blacklist_input.endsWith('.fasta')) { + ch_blacklist_fasta = ch_blacklist_input + } else { // TODO optionally convert bed to fasta + error "The blacklist file must be in fasta nucleotide format. \n\tBlacklist: ${params.blacklist}" + } + ch_blacklist_index = BWA_INDEX_BL(ch_blacklist_fasta).index + ch_reference_index = BWA_INDEX_REF(ch_fasta).index KHMER_UNIQUEKMERS(ch_fasta, params.read_length) + ch_gsize = KHMER_UNIQUEKMERS.out.kmers.map { it.text.trim() } + ch_gene_bed = GTF2BED ( file(params.gtf) ).bed + //ch_chrom_sizes = CUSTOM_GETCHROMSIZES ( ch_fasta ).sizes + SPLIT_REF_CHROMS(ch_fasta) + ch_chrom_sizes = SPLIT_REF_CHROMS.out.chrom_sizes + ch_chrom_files = SPLIT_REF_CHROMS.out.chrom_files + ch_bwa_index = BWA_INDEX ( ch_fasta ).index + ch_bowtie2_index = BOWTIE2_BUILD ( ch_fasta ).index } else { Channel.fromPath(params.genomes[ params.genome ].blacklist_files, checkIfExists: true) .collect() - .set{ ch_blacklist_files } + .set{ ch_blacklist_index } Channel.fromPath(params.genomes[ params.genome ].reference_files, checkIfExists: true) .collect() - .set{ ch_reference_files } + .set{ ch_reference_index } Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) .set{ ch_chrom_sizes } Channel.fromPath("${params.genomes[ params.genome ].chromosomes_dir}", type: 'dir', checkIfExists: true) @@ -27,15 +41,14 @@ workflow PREPARE_GENOME { checkIfExists: true) .set{ ch_gene_info } Channel.value(params.genomes[ params.genome ].effective_genome_size) - .set{ ch_egs} + .set{ ch_gsize} } emit: - blacklist_files = ch_blacklist_files - blacklist_name = ch_blacklist_name - reference_files = ch_reference_files + blacklist_index = ch_blacklist_index + reference_index = ch_reference_index chrom_sizes = ch_chrom_sizes chrom_files = ch_chrom_files gene_info = ch_gene_info - effective_genome_size = ch_egs + effective_genome_size = ch_gsize } From b23c733a2f1bd578615639ebfd8da0d37a63f3ae Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 3 Oct 2023 23:19:57 -0400 Subject: [PATCH 044/100] refactor: rename bl/ref files to index list --- main.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/main.nf b/main.nf index 8ff04754..f5c788d8 100644 --- a/main.nf +++ b/main.nf @@ -47,8 +47,8 @@ workflow { chrom_sizes = PREPARE_GENOME.out.chrom_sizes effective_genome_size = PREPARE_GENOME.out.effective_genome_size - ALIGN_BLACKLIST(trimmed_fastqs, PREPARE_GENOME.out.blacklist_files, PREPARE_GENOME.out.blacklist_name) | BAM_TO_FASTQ - ALIGN_GENOME(BAM_TO_FASTQ.out.reads, PREPARE_GENOME.out.reference_files) + ALIGN_BLACKLIST(trimmed_fastqs, PREPARE_GENOME.out.blacklist_index, PREPARE_GENOME.out.blacklist_name) | BAM_TO_FASTQ + ALIGN_GENOME(BAM_TO_FASTQ.out.reads, PREPARE_GENOME.out.reference_index) ALIGN_GENOME.out.bam.set{ aligned_bam } DEDUPLICATE(aligned_bam, chrom_sizes, effective_genome_size) From b248ef6e10347b5f3b6003eeb914b1f81cc69309 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 08:29:21 -0400 Subject: [PATCH 045/100] feat: use bedtools getfasta --- modules.json | 5 +++ modules/nf-core/bedtools/getfasta/main.nf | 37 ++++++++++++++++++++++ modules/nf-core/bedtools/getfasta/meta.yml | 34 ++++++++++++++++++++ 3 files changed, 76 insertions(+) create mode 100644 modules/nf-core/bedtools/getfasta/main.nf create mode 100644 modules/nf-core/bedtools/getfasta/meta.yml diff --git a/modules.json b/modules.json index cbdfea8a..09ad9474 100644 --- a/modules.json +++ b/modules.json @@ -26,6 +26,11 @@ "https://github.com/nf-core/modules.git": { "modules": { "nf-core": { + "bedtools/getfasta": { + "branch": "master", + "git_sha": "911696ea0b62df80e900ef244d7867d177971f73", + "installed_by": ["modules"] + }, "bwa/index": { "branch": "master", "git_sha": "bfed129da5134b4439b1821c917972570d44d39c", diff --git a/modules/nf-core/bedtools/getfasta/main.nf b/modules/nf-core/bedtools/getfasta/main.nf new file mode 100644 index 00000000..84adc4c9 --- /dev/null +++ b/modules/nf-core/bedtools/getfasta/main.nf @@ -0,0 +1,37 @@ +process BEDTOOLS_GETFASTA { + tag "$bed" + label 'process_single' + + conda "bioconda::bedtools=2.30.0" + container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? + 'https://depot.galaxyproject.org/singularity/bedtools:2.30.0--hc088bd4_0' : + 'biocontainers/bedtools:2.30.0--hc088bd4_0' }" + + input: + path bed + path fasta + + output: + path "*.fa" , emit: fasta + path "versions.yml" , emit: versions + + when: + task.ext.when == null || task.ext.when + + script: + def args = task.ext.args ?: '' + def prefix = task.ext.prefix ?: "${bed.baseName}" + """ + bedtools \\ + getfasta \\ + $args \\ + -fi $fasta \\ + -bed $bed \\ + -fo ${prefix}.fa + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + bedtools: \$(bedtools --version | sed -e "s/bedtools v//g") + END_VERSIONS + """ +} diff --git a/modules/nf-core/bedtools/getfasta/meta.yml b/modules/nf-core/bedtools/getfasta/meta.yml new file mode 100644 index 00000000..11335100 --- /dev/null +++ b/modules/nf-core/bedtools/getfasta/meta.yml @@ -0,0 +1,34 @@ +name: bedtools_getfasta +description: extract sequences in a FASTA file based on intervals defined in a feature file. +keywords: + - bed + - fasta + - getfasta +tools: + - bedtools: + description: | + A set of tools for genomic analysis tasks, specifically enabling genome arithmetic (merge, count, complement) on various file types. + documentation: https://bedtools.readthedocs.io/en/latest/content/tools/intersect.html + licence: ["MIT"] +input: + - bed: + type: file + description: Bed feature file + pattern: "*.{bed}" + - fasta: + type: file + description: Input fasta file + pattern: "*.{fa,fasta}" + +output: + - fasta: + type: file + description: Output fasta file with extracted sequences + pattern: "*.{fa}" + - versions: + type: file + description: File containing software versions + pattern: "versions.yml" +authors: + - "@joseespinosa" + - "@drpatelh" From e0a49fa3ab820839753d85d0b459708c5c6d1992 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 10:09:43 -0400 Subject: [PATCH 046/100] fix: make splitRef.py executable --- bin/splitRef.py | 0 1 file changed, 0 insertions(+), 0 deletions(-) mode change 100644 => 100755 bin/splitRef.py diff --git a/bin/splitRef.py b/bin/splitRef.py old mode 100644 new mode 100755 From d34ea9edb30f1c4f0cf6b9585e6302d7c3f4762e Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 10:19:11 -0400 Subject: [PATCH 047/100] feat: create config file for custom genome --- modules/local/prepare_genome.nf | 43 +++++++++++++++++++++++++++++++-- 1 file changed, 41 insertions(+), 2 deletions(-) diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index a9e37a8b..2db8006f 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -2,13 +2,13 @@ process GTF2BED { tag { gtf } label 'process_single' - container 'quay.io/biocontainers/bedops:2.4.41--h4ac6f70_1' + container 'nciccbr/ccbr_ubuntu_base_20.04:v5' input: path(gtf) output: - path("${gtf.baseName}.bed") + path("${gtf.baseName}.bed"), emit: bed script: """ @@ -40,3 +40,42 @@ process SPLIT_REF_CHROMS { touch ${fasta.baseName}.chrom.sizes """ } + +process CREATE_GENOME_CONFIG { + label 'process_single' + container "${params.containers.base}" + + input: + tuple val(meta_ref), path(reference_index) + tuple val(meta_bl), path(blacklist_index) + path(chrom_sizes) + path(chrom_files) + path(gene_info) + val(effective_genome_size) + + output: + path('genome/') + + script: + def genome_name = 'custom_genome' // TODO allow user to set this + """ + #!/usr/bin/env python + import os + import pprint + + os.mkdir('genome/') + genome = dict(reference_index = "${reference_index}", + blacklist_index = "${blacklist_index}", + effective_genome_size = "${effective_genome_size}", + chrom_sizes = "${chrom_sizes}", + gene_info = "${gene_info}", + chromosomes_dir = "${chrom_files}" + ) + print(genome) + with open('genome/${genome_name}.config', 'w') as conf_file: + conf_file.write("'${genome_name}' {") + for k, v in genome: + conf_file.write(f"\t{k} = \${{params.index_dir}}/{v}") + conf_file.write("}") + """ +} From cdd9f4c39c16eb84ffb5342ea83f1dcd80cdc381 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 10:55:45 -0400 Subject: [PATCH 048/100] fix: create output dir --- bin/splitRef.py | 1 + 1 file changed, 1 insertion(+) diff --git a/bin/splitRef.py b/bin/splitRef.py index b5d9bed4..5bac3c6a 100755 --- a/bin/splitRef.py +++ b/bin/splitRef.py @@ -42,6 +42,7 @@ def parsed(filename): def main(fasta_fn, chrom_sizes_fn, outdir): + os.mkdir(outdir) chromsizesfh = open(chrom_sizes_fn, "w") for chrom, seq, chromsize in parsed(fasta_fn): From 65b6147c657dd4e54e3e80fba8322b0b6e30defa Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 10:56:19 -0400 Subject: [PATCH 049/100] feat: copy custom genome files --- conf/modules.config | 6 ++++++ modules/local/prepare_genome.nf | 37 ++++++++++++++++++++------------- 2 files changed, 29 insertions(+), 14 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 2496842b..5723e2bc 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -24,4 +24,10 @@ process { '-b file:/opt2/TruSeq_and_nextera_adapters.consolidated.fa' ].join(' ').trim() } + withName: 'WRITE_GENOME_CONFIG' { + publishDir = [ + path: { "${params.outdir}/genome" }, + mode: "copy" + ] + } } diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index 2db8006f..f45fdfc7 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -41,7 +41,7 @@ process SPLIT_REF_CHROMS { """ } -process CREATE_GENOME_CONFIG { +process WRITE_GENOME_CONFIG { label 'process_single' container "${params.containers.base}" @@ -54,28 +54,37 @@ process CREATE_GENOME_CONFIG { val(effective_genome_size) output: - path('genome/') + path("*.config"), emit: conf + path("custom_genome/*"), emit: files script: - def genome_name = 'custom_genome' // TODO allow user to set this + def genome_name = 'custom_genome' """ #!/usr/bin/env python import os import pprint + import shutil + os.makedirs("${genome_name}/") + for subdir, filelist in (('reference/', "${reference_index}"), ('blacklist', "${blacklist_index}"), ('chroms', "${chrom_files}")): + dirpath = f"${{genome_name}}/{subdir}" + os.mkdir(dirpath) + for file in filelist.split(): + shutil.copy(file, dirpath) + for file in ("${chrom_sizes}", "${gene_info}"): + shutil.copy(file, "${genome_name}/") - os.mkdir('genome/') - genome = dict(reference_index = "${reference_index}", - blacklist_index = "${blacklist_index}", - effective_genome_size = "${effective_genome_size}", - chrom_sizes = "${chrom_sizes}", - gene_info = "${gene_info}", - chromosomes_dir = "${chrom_files}" + genome = dict(reference_index = "\${params.index_dir}/${genome_name}/reference/*", + blacklist_index = "\${params.index_dir}/${genome_name}/blacklist/*", + effective_genome_size = "${effective_genome_size}", + chrom_sizes = "\${params.index_dir}/${genome_name}/${chrom_sizes}", + gene_info = "\${params.index_dir}/${genome_name}/${gene_info}", + chromosomes_dir = "\${params.index_dir}/${genome_name}/chroms/*" ) - print(genome) - with open('genome/${genome_name}.config', 'w') as conf_file: + pprint.pprint(genome) + with open('${genome_name}.config', 'w') as conf_file: conf_file.write("'${genome_name}' {") - for k, v in genome: - conf_file.write(f"\t{k} = \${{params.index_dir}}/{v}") + for k, v in genome.items(): + conf_file.write(f"\\t{k} = {v}\\n") conf_file.write("}") """ } From fe3c9760e79414e0f3ec05641457fd48c8d526a0 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 10:56:55 -0400 Subject: [PATCH 050/100] feat: use improved blacklist --- conf/genomes.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/genomes.config b/conf/genomes.config index e75b2404..28b6ce46 100644 --- a/conf/genomes.config +++ b/conf/genomes.config @@ -1,7 +1,7 @@ params { genomes { 'hg38' { - blacklist_index = "${params.index_dir}/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.*" + blacklist_index = "${params.index_dir}/hg38_basic/indexes/blacklist/*" reference_index = "${params.index_dir}/hg38_basic/indexes/hg38*" effective_genome_size = 2700000000 chrom_sizes = "${params.index_dir}/hg38_basic/indexes/hg38.fa.sizes" From acf2e0f9fbcff9e95ba62cc686a8234aa757fd3a Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 10:57:20 -0400 Subject: [PATCH 051/100] test: use human blacklist for test dataset --- conf/test.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test.config b/conf/test.config index 67ea9203..1c1aeb28 100644 --- a/conf/test.config +++ b/conf/test.config @@ -10,7 +10,7 @@ params { // Genome references genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' - blacklist = '' + blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' genome = 'custom_test' save_reference = true // https://github.com/nf-core/chipseq/blob/51eba00b32885c4d0bec60db3cb0a45eb61e34c5/subworkflows/local/prepare_genome.nf#L47-L51 From d78437df3f6bc2283909e5a2595c30c2074f79fb Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 11:00:06 -0400 Subject: [PATCH 052/100] fix: glob bwa output dir --- modules.json | 2 +- modules/CCBR/bwa/index/main.nf | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/modules.json b/modules.json index 09ad9474..14ccc61a 100644 --- a/modules.json +++ b/modules.json @@ -7,7 +7,7 @@ "CCBR": { "bwa/index": { "branch": "main", - "git_sha": "b39dceb9d9dd55804f8cd81181a315a48dc0f4c6", + "git_sha": "a4830988cb507ff060bf45fbac2e81f72413ab53", "installed_by": ["modules"] }, "bwa/mem": { diff --git a/modules/CCBR/bwa/index/main.nf b/modules/CCBR/bwa/index/main.nf index a30f3d76..16188ff0 100644 --- a/modules/CCBR/bwa/index/main.nf +++ b/modules/CCBR/bwa/index/main.nf @@ -8,7 +8,7 @@ process BWA_INDEX { tuple val(meta), path(fasta) output: - tuple val(meta), path(bwa) , emit: index + tuple val(meta), path("bwa/*") , emit: index path "versions.yml" , emit: versions when: From bc6f59b4ddf6dbb9bee901a42e6890d54adc0a69 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 11:00:38 -0400 Subject: [PATCH 053/100] refactor: make align blacklist use bwa modules --- main.nf | 6 ++++-- modules/local/align.nf | 16 +++++++--------- 2 files changed, 11 insertions(+), 11 deletions(-) diff --git a/main.nf b/main.nf index f5c788d8..8e7542c6 100644 --- a/main.nf +++ b/main.nf @@ -28,7 +28,8 @@ include { CALL_PEAKS } from './subworkflows/local/peaks.nf' // MODULES include { CUTADAPT } from "./modules/CCBR/cutadapt" -include { ALIGN_BLACKLIST +include { BWA_MEM as ALIGN_BLACKLIST } from "./modules/CCBR/bwa/mem" +include { FILTER_BLACKLIST BAM_TO_FASTQ ALIGN_GENOME } from "./modules/local/align.nf" include { PHANTOM_PEAKS } from "./modules/local/qc.nf" @@ -47,7 +48,8 @@ workflow { chrom_sizes = PREPARE_GENOME.out.chrom_sizes effective_genome_size = PREPARE_GENOME.out.effective_genome_size - ALIGN_BLACKLIST(trimmed_fastqs, PREPARE_GENOME.out.blacklist_index, PREPARE_GENOME.out.blacklist_name) | BAM_TO_FASTQ + ALIGN_BLACKLIST(trimmed_fastqs, PREPARE_GENOME.out.blacklist_index) + ALIGN_BLACKLIST.out.bam | FILTER_BLACKLIST | BAM_TO_FASTQ ALIGN_GENOME(BAM_TO_FASTQ.out.reads, PREPARE_GENOME.out.reference_index) ALIGN_GENOME.out.bam.set{ aligned_bam } diff --git a/modules/local/align.nf b/modules/local/align.nf index f47f8b22..d88484ce 100644 --- a/modules/local/align.nf +++ b/modules/local/align.nf @@ -1,5 +1,5 @@ -process ALIGN_BLACKLIST { // TODO: refactor this as a subworkflow that combines alignment and filtering processes +process FILTER_BLACKLIST { // TODO: refactor this as a subworkflow that combines alignment and filtering processes tag { meta.id } label 'align' label 'process_higher' @@ -7,9 +7,7 @@ process ALIGN_BLACKLIST { // TODO: refactor this as a subworkflow that combines container = "${params.containers.base}" input: - tuple val(meta), path(fastq) - path(index_files) - val(index_name) + tuple val(meta), path(bam) output: tuple val(meta), path("${meta.id}.no_blacklist.bam"), emit: bam @@ -18,8 +16,6 @@ process ALIGN_BLACKLIST { // TODO: refactor this as a subworkflow that combines def prefix = task.ext.prefix ?: "${meta.id}" def filter_flag = meta.single_end ? '4' : '12' """ - bwa mem -t ${task.cpus} ${index_name} ${fastq} > ${prefix}.sam - samtools view \\ -@ ${task.cpus} \\ -f${filter_flag} \\ @@ -66,7 +62,7 @@ process BAM_TO_FASTQ { """ } -process ALIGN_GENOME { +process ALIGN_GENOME { // TODO refactor as subworkflow tag { meta.id } label 'align' label 'process_higher' @@ -75,7 +71,7 @@ process ALIGN_GENOME { input: tuple val(meta), path(fastq) - path(reference_files) + tuple val(meta_ref), path(reference_files) output: tuple val(meta), path("*.aligned.filtered.bam"), emit: bam @@ -104,7 +100,9 @@ process ALIGN_GENOME { mkdir \$TMP trap 'rm -rf "\$TMP"' EXIT - bwa mem -t ${task.cpus} ${params.genome} ${fastq} > ${prefix}.bam + INDEX=`find -L ./ -name "*.amb" | sed 's/\\.amb\$//'` + + bwa mem -t ${task.cpus} \$INDEX ${fastq} > ${prefix}.bam samtools sort \\ -@ ${task.cpus} \\ -m 2G \\ From ba2c6fe53a85baebc2297f1141af46d2b4966fd5 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 11:01:01 -0400 Subject: [PATCH 054/100] feat: succcessfully prepare custom reference genome --- subworkflows/local/prepare_genome.nf | 32 ++++++++++++++++++++-------- 1 file changed, 23 insertions(+), 9 deletions(-) diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index ff08e5ec..0d93f60f 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -1,30 +1,44 @@ include { BWA_INDEX as BWA_INDEX_BL BWA_INDEX as BWA_INDEX_REF } from "../../modules/CCBR/bwa/index" include { KHMER_UNIQUEKMERS } from '../../modules/nf-core/khmer/uniquekmers' +include { BEDTOOLS_GETFASTA } from '../../modules/nf-core/bedtools/getfasta/main' include { SPLIT_REF_CHROMS } from '../../modules/local/prepare_genome.nf' include { GTF2BED } from '../../modules/local/prepare_genome.nf' +include { WRITE_GENOME_CONFIG } from "../../modules/local/prepare_genome.nf" workflow PREPARE_GENOME { main: if (params.genome_fasta && params.genes_gtf) { ch_fasta = file(params.genome_fasta) ch_blacklist_input = file(params.blacklist) - if (ch_blacklist_input.endsWith('.fa') || ch_blacklist_input.endsWith('.fna') || ch_blacklist_input.endsWith('.fasta')) { + if (ch_blacklist_input.endsWith('.bed')) { + BEDTOOLS_GETFASTA(ch_blacklist_input, ch_fasta) + ch_blacklist_fasta = BEDTOOLS_GETFASTA.out.fasta + } else { ch_blacklist_fasta = ch_blacklist_input - } else { // TODO optionally convert bed to fasta - error "The blacklist file must be in fasta nucleotide format. \n\tBlacklist: ${params.blacklist}" } - ch_blacklist_index = BWA_INDEX_BL(ch_blacklist_fasta).index - ch_reference_index = BWA_INDEX_REF(ch_fasta).index + ch_blacklist_index = BWA_INDEX_BL([['id': 'blacklist'], ch_blacklist_fasta]).index + ch_reference_index = BWA_INDEX_REF([['id': 'genome'], ch_fasta]).index KHMER_UNIQUEKMERS(ch_fasta, params.read_length) ch_gsize = KHMER_UNIQUEKMERS.out.kmers.map { it.text.trim() } - ch_gene_bed = GTF2BED ( file(params.gtf) ).bed - //ch_chrom_sizes = CUSTOM_GETCHROMSIZES ( ch_fasta ).sizes + ch_gene_info = GTF2BED ( file(params.genes_gtf) ).bed SPLIT_REF_CHROMS(ch_fasta) ch_chrom_sizes = SPLIT_REF_CHROMS.out.chrom_sizes ch_chrom_files = SPLIT_REF_CHROMS.out.chrom_files - ch_bwa_index = BWA_INDEX ( ch_fasta ).index - ch_bowtie2_index = BOWTIE2_BUILD ( ch_fasta ).index + + // save reference files + if (params.save_reference) { + //file("${params.outdir}/genome/").mkdirs() + WRITE_GENOME_CONFIG( + ch_reference_index, + ch_blacklist_index, + ch_chrom_sizes, + ch_chrom_files, + ch_gene_info, + ch_gsize + ) + + } } else { Channel.fromPath(params.genomes[ params.genome ].blacklist_files, checkIfExists: true) From 611ef4c1473b6d4c1d6b859d4055140cbffc96f9 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 12:34:22 -0400 Subject: [PATCH 055/100] feat: optionally turn peak callers on/off --- nextflow.config | 8 ++++++++ subworkflows/local/peaks.nf | 31 +++++++++++++++++++------------ 2 files changed, 27 insertions(+), 12 deletions(-) diff --git a/nextflow.config b/nextflow.config index cc55c7e4..6e7c5832 100644 --- a/nextflow.config +++ b/nextflow.config @@ -57,10 +57,18 @@ params { cutoff = 0.01 } } + sicer { + species = "sacCer1" // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py + } + run { // some steps can be turned on/off for debugging purposes qc = true normalize_input = true call_peaks = true + gem = true + sicer = true + macs_broad = true + macs_narrow = true } } diff --git a/subworkflows/local/peaks.nf b/subworkflows/local/peaks.nf index 434f7a27..ab82e42e 100644 --- a/subworkflows/local/peaks.nf +++ b/subworkflows/local/peaks.nf @@ -20,7 +20,7 @@ include { BAM_TO_BED } from "../../modules/local/bedtools.nf" workflow CALL_PEAKS { take: chrom_sizes - chrom_files + chrom_dir deduped_tagalign deduped_bam frag_lengths @@ -74,20 +74,27 @@ workflow CALL_PEAKS { ch_tagalign .combine(Channel.fromPath(params.gem.read_dists, checkIfExists: true)) .combine(chrom_sizes) - .combine(chrom_files) + .combine(chrom_dir) .combine(effective_genome_size) .set { ch_gem } - ch_macs | MACS_BROAD - ch_macs | MACS_NARROW - ch_sicer | SICER | CONVERT_SICER - GEM(ch_gem) - - CONVERT_SICER.out.peak - .mix(GEM.out.peak, - MACS_BROAD.out.peak, - MACS_NARROW.out.peak - ).set{ ch_peaks } + ch_peaks = Channel.empty() + if (params.run.macs_broad) { + ch_macs | MACS_BROAD + ch_peaks = ch_peaks.mix(MACS_BROAD.out.peak) + } + if (params.run.macs_narrow) { + ch_macs | MACS_NARROW + ch_peaks = ch_peaks.mix(MACS_NARROW.out.peak) + } + if (params.run.sicer) { + ch_sicer | SICER | CONVERT_SICER + ch_peaks = ch_peaks.mix(CONVERT_SICER.out.peak) + } + if (params.run.gem) { + ch_gem | GEM + ch_peaks = ch_peaks.mix(GEM.out.peak) + } // Create Channel with meta, deduped bam, peak file, peak-calling tool, and chrom sizes fasta deduped_bam.cross(ch_peaks) From f6f771f9e3f98e85d03155e92aa03a5c4cff9e59 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 12:35:00 -0400 Subject: [PATCH 056/100] fix: sicer supports a limited number of species see https://github.com/CCBR/CHAMPAGNE/issues/109 --- modules/local/peaks.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/peaks.nf b/modules/local/peaks.nf index 334937e9..16c7306d 100644 --- a/modules/local/peaks.nf +++ b/modules/local/peaks.nf @@ -119,7 +119,7 @@ process SICER { sicer \\ -t ${chip} \\ -c ${input} \\ - -s ${params.genome} \\ + -s ${params.sicer.species} \\ -rt 100 \\ -w 300 \\ -f ${fraglen} \\ From f4abb786b260b467e8580684de1fb6928d091882 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 12:35:38 -0400 Subject: [PATCH 057/100] fix: make sure split_ref_chroms outputs directory --- main.nf | 2 +- modules/local/prepare_genome.nf | 9 ++++++--- subworkflows/local/prepare_genome.nf | 8 ++++---- 3 files changed, 11 insertions(+), 8 deletions(-) diff --git a/main.nf b/main.nf index 8e7542c6..ebdc0aed 100644 --- a/main.nf +++ b/main.nf @@ -80,7 +80,7 @@ workflow { } if (params.run.call_peaks) { - CALL_PEAKS(chrom_sizes, PREPARE_GENOME.out.chrom_files, deduped_tagalign, deduped_bam, frag_lengths, effective_genome_size) + CALL_PEAKS(chrom_sizes, PREPARE_GENOME.out.chrom_dir, deduped_tagalign, deduped_bam, frag_lengths, effective_genome_size) ch_multiqc = ch_multiqc.mix(CALL_PEAKS.out.plots) } diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index f45fdfc7..a97a707e 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -29,7 +29,7 @@ process SPLIT_REF_CHROMS { output: path("${fasta.baseName}.chrom.sizes"), emit: chrom_sizes - path("chroms/*") , emit: chrom_files + path("chroms/") , emit: chrom_dir script: """ @@ -38,6 +38,8 @@ process SPLIT_REF_CHROMS { stub: """ touch ${fasta.baseName}.chrom.sizes + mkdir -p chroms/ + touch chroms/chr1.fa """ } @@ -49,7 +51,7 @@ process WRITE_GENOME_CONFIG { tuple val(meta_ref), path(reference_index) tuple val(meta_bl), path(blacklist_index) path(chrom_sizes) - path(chrom_files) + path(chrom_dir) path(gene_info) val(effective_genome_size) @@ -65,11 +67,12 @@ process WRITE_GENOME_CONFIG { import pprint import shutil os.makedirs("${genome_name}/") - for subdir, filelist in (('reference/', "${reference_index}"), ('blacklist', "${blacklist_index}"), ('chroms', "${chrom_files}")): + for subdir, filelist in (('reference/', "${reference_index}"), ('blacklist', "${blacklist_index}")): dirpath = f"${{genome_name}}/{subdir}" os.mkdir(dirpath) for file in filelist.split(): shutil.copy(file, dirpath) + shutil.copytree("${chrom_dir}", '${genome_name}/chroms/') for file in ("${chrom_sizes}", "${gene_info}"): shutil.copy(file, "${genome_name}/") diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index 0d93f60f..10d52700 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -24,7 +24,7 @@ workflow PREPARE_GENOME { ch_gene_info = GTF2BED ( file(params.genes_gtf) ).bed SPLIT_REF_CHROMS(ch_fasta) ch_chrom_sizes = SPLIT_REF_CHROMS.out.chrom_sizes - ch_chrom_files = SPLIT_REF_CHROMS.out.chrom_files + ch_chrom_dir = SPLIT_REF_CHROMS.out.chrom_dir // save reference files if (params.save_reference) { @@ -33,7 +33,7 @@ workflow PREPARE_GENOME { ch_reference_index, ch_blacklist_index, ch_chrom_sizes, - ch_chrom_files, + ch_chrom_dir, ch_gene_info, ch_gsize ) @@ -50,7 +50,7 @@ workflow PREPARE_GENOME { Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) .set{ ch_chrom_sizes } Channel.fromPath("${params.genomes[ params.genome ].chromosomes_dir}", type: 'dir', checkIfExists: true) - .set{ ch_chrom_files } + .set{ ch_chrom_dir } Channel.fromPath(params.genomes[ params.genome ].gene_info, checkIfExists: true) .set{ ch_gene_info } @@ -62,7 +62,7 @@ workflow PREPARE_GENOME { blacklist_index = ch_blacklist_index reference_index = ch_reference_index chrom_sizes = ch_chrom_sizes - chrom_files = ch_chrom_files + chrom_dir = ch_chrom_dir gene_info = ch_gene_info effective_genome_size = ch_gsize } From 1db029201fe88c44574c89b3b401231c474b326b Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 12:53:03 -0400 Subject: [PATCH 058/100] feat: only run call_peaks if any peak caller run param is true --- conf/ci_stub.config | 10 +++++++++- conf/test.config | 11 +++++++++-- main.nf | 3 +-- 3 files changed, 19 insertions(+), 5 deletions(-) diff --git a/conf/ci_stub.config b/conf/ci_stub.config index a5f58fd7..42a58baf 100644 --- a/conf/ci_stub.config +++ b/conf/ci_stub.config @@ -19,6 +19,15 @@ params { db_dir = "tests/data/" } multiqc_config = "assets/multiqc_config.yaml" + run { + qc = true + normalize_input = false + call_peaks = true + gem = false + sicer = false + macs_broad = true + macs_narrow = true + } genomes { 'test' { // blank files for testing stubs on GitHub Actions blacklist = 'test.blacklist' @@ -30,7 +39,6 @@ params { chromosomes_dir = "tests/data/chroms/" } } - } process { diff --git a/conf/test.config b/conf/test.config index 1c1aeb28..755b1aba 100644 --- a/conf/test.config +++ b/conf/test.config @@ -11,7 +11,7 @@ params { genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' - genome = 'custom_test' + genome = 'sacCer3' save_reference = true // https://github.com/nf-core/chipseq/blob/51eba00b32885c4d0bec60db3cb0a45eb61e34c5/subworkflows/local/prepare_genome.nf#L47-L51 max_cpus = 32 // for interactive node on biowulf @@ -21,7 +21,14 @@ params { run { qc = false normalize_input = false - call_peaks = false + call_peaks = true + gem = true + sicer = false + macs_broad = true + macs_narrow = true + } + sicer { + species = "sacCer1" } } dag { diff --git a/main.nf b/main.nf index ebdc0aed..bcf6b8c3 100644 --- a/main.nf +++ b/main.nf @@ -78,8 +78,7 @@ workflow { ch_multiqc = ch_multiqc.mix(QC.out.multiqc_input) } - - if (params.run.call_peaks) { + if (params.run.call_peaks && [params.run.macs_broad, params.run.macs_narrow, params.run.gem, params.run.sicer].any()) { CALL_PEAKS(chrom_sizes, PREPARE_GENOME.out.chrom_dir, deduped_tagalign, deduped_bam, frag_lengths, effective_genome_size) ch_multiqc = ch_multiqc.mix(CALL_PEAKS.out.plots) } From 37e130ac006a7f9fc99a1a5f605a5be50336e580 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 12:55:22 -0400 Subject: [PATCH 059/100] fix: config for test stubs --- conf/ci_stub.config | 15 ++------------- 1 file changed, 2 insertions(+), 13 deletions(-) diff --git a/conf/ci_stub.config b/conf/ci_stub.config index 42a58baf..078c0b7e 100644 --- a/conf/ci_stub.config +++ b/conf/ci_stub.config @@ -18,21 +18,10 @@ params { conf = "assets/fastq_screen_biowulf.conf" db_dir = "tests/data/" } - multiqc_config = "assets/multiqc_config.yaml" - run { - qc = true - normalize_input = false - call_peaks = true - gem = false - sicer = false - macs_broad = true - macs_narrow = true - } genomes { 'test' { // blank files for testing stubs on GitHub Actions - blacklist = 'test.blacklist' - blacklist_files = "tests/data/test.blacklist" - reference_files = "tests/data/test/*" + blacklist_index = "tests/data/test.blacklist" + reference_index = "tests/data/test/*" effective_genome_size = 2700000000 chrom_sizes = "tests/data/test.fa.sizes" gene_info = "tests/data/geneinfo.bed" From 208e055bde6f370f5395fe363d11d21f03d9ea57 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 13:23:25 -0400 Subject: [PATCH 060/100] fix: new key names for blacklist and ref index --- subworkflows/local/prepare_genome.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index 10d52700..65fa1170 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -41,10 +41,10 @@ workflow PREPARE_GENOME { } } else { - Channel.fromPath(params.genomes[ params.genome ].blacklist_files, checkIfExists: true) + Channel.fromPath(params.genomes[ params.genome ].blacklist_index, checkIfExists: true) .collect() .set{ ch_blacklist_index } - Channel.fromPath(params.genomes[ params.genome ].reference_files, checkIfExists: true) + Channel.fromPath(params.genomes[ params.genome ].reference_index, checkIfExists: true) .collect() .set{ ch_reference_index } Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) From d849c68b7f9a1975fc9be4e371366a8512880000 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 13:23:46 -0400 Subject: [PATCH 061/100] fix: set custom genome options --- conf/test_paired.config | 17 +++++++++++++++-- nextflow.config | 8 ++++++-- 2 files changed, 21 insertions(+), 4 deletions(-) diff --git a/conf/test_paired.config b/conf/test_paired.config index 58688992..585a059a 100644 --- a/conf/test_paired.config +++ b/conf/test_paired.config @@ -5,9 +5,15 @@ params { outdir = 'results/test_paired' input = 'assets/samplesheet_test_paired.csv' // adapted from 'https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/samplesheet/v2.0/samplesheet_test.csv' - genome = 'hg38' + genome = 'sacCer3' read_length = 50 + // Genome references + genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' + genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' + blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' + save_reference = true + max_cpus = 32 // for interactive node on biowulf max_memory = '120 GB' // for interactive node on biowulf @@ -15,7 +21,14 @@ params { run { qc = false normalize_input = false - call_peaks = false // no peaks exist with this yeast dataset with hg38 as the reference + call_peaks = true + gem = false + sicer = false + macs_broad = true + macs_narrow = true + } + sicer { + species = "sacCer1" } } dag { diff --git a/nextflow.config b/nextflow.config index 6e7c5832..6a48d0c4 100644 --- a/nextflow.config +++ b/nextflow.config @@ -5,8 +5,12 @@ params { seq_center = null read_length = null genome = null - fasta = null - gtf = null + + // custom genome options + genome_fasta = null + genes_gtf = null + blacklist = null + save_reference = false // Boilerplate options outdir = 'results' From 5187d3eac290c3ff8d7c2fc0df81330669876677 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 14:59:25 -0400 Subject: [PATCH 062/100] feat: make sure custom genome config is 'roundtrippable' can be reused with '-c path/to/custom.config' --- modules/local/prepare_genome.nf | 42 +++++++++++++----- subworkflows/local/prepare_genome.nf | 65 +++++++++++++--------------- 2 files changed, 63 insertions(+), 44 deletions(-) diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index a97a707e..87c30022 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -1,3 +1,14 @@ +process EMIT_META { + input: + path(file) + output: + tuple val(file.baseName), path(file) + script: + """ + echo $file + """ +} + process GTF2BED { tag { gtf } label 'process_single' @@ -47,6 +58,10 @@ process WRITE_GENOME_CONFIG { label 'process_single' container "${params.containers.base}" + publishDir = [ + path: { "${params.outdir}/genome" }, + mode: "copy" + ] input: tuple val(meta_ref), path(reference_index) tuple val(meta_bl), path(blacklist_index) @@ -57,7 +72,7 @@ process WRITE_GENOME_CONFIG { output: path("*.config"), emit: conf - path("custom_genome/*"), emit: files + path("custom_genome/"), emit: files // TODO can't use genome_name variable here, nextflow thinks it's null?? script: def genome_name = 'custom_genome' @@ -76,18 +91,25 @@ process WRITE_GENOME_CONFIG { for file in ("${chrom_sizes}", "${gene_info}"): shutil.copy(file, "${genome_name}/") - genome = dict(reference_index = "\${params.index_dir}/${genome_name}/reference/*", - blacklist_index = "\${params.index_dir}/${genome_name}/blacklist/*", - effective_genome_size = "${effective_genome_size}", - chrom_sizes = "\${params.index_dir}/${genome_name}/${chrom_sizes}", - gene_info = "\${params.index_dir}/${genome_name}/${gene_info}", - chromosomes_dir = "\${params.index_dir}/${genome_name}/chroms/*" + genome = dict(reference_index = '"\${params.index_dir}/${genome_name}/reference/*"', + blacklist_index = '"\${params.index_dir}/${genome_name}/blacklist/*"', + chromosomes_dir = '"\${params.index_dir}/${genome_name}/chroms/"', + chrom_sizes = '"\${params.index_dir}/${genome_name}/${chrom_sizes}"', + gene_info = '"\${params.index_dir}/${genome_name}/${gene_info}"', + effective_genome_size = "${effective_genome_size}" ) pprint.pprint(genome) with open('${genome_name}.config', 'w') as conf_file: - conf_file.write("'${genome_name}' {") + head = ["params {\\n", + '\\tindex_dir = "${params.outdir}/genome/"\\n', + "\\tgenomes {\\n" + "\\t\\t'${genome_name}' {\\n"] + conf_file.writelines(head) for k, v in genome.items(): - conf_file.write(f"\\t{k} = {v}\\n") - conf_file.write("}") + conf_file.write(f'\\t\\t\\t{k} = {v}\\n') + tail = ["\\t\\t}\\n", + "\\t}\\n", + "}\\n"] + conf_file.writelines(tail) """ } diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index 65fa1170..e13771d5 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -5,57 +5,54 @@ include { BEDTOOLS_GETFASTA } from '../../modules/nf-core/bedtools/getf include { SPLIT_REF_CHROMS } from '../../modules/local/prepare_genome.nf' include { GTF2BED } from '../../modules/local/prepare_genome.nf' include { WRITE_GENOME_CONFIG } from "../../modules/local/prepare_genome.nf" +include { EMIT_META as EMIT_META_BL + EMIT_META as EMIT_META_REF } from "../../modules/local/prepare_genome.nf" workflow PREPARE_GENOME { main: - if (params.genome_fasta && params.genes_gtf) { - ch_fasta = file(params.genome_fasta) - ch_blacklist_input = file(params.blacklist) + if (params.genomes[ params.genome ]) { + println "Using ${params.genome} as the reference" + + ch_blacklist_index = Channel.fromPath(params.genomes[ params.genome ].blacklist_index, checkIfExists: true) + .collect() | EMIT_META_BL + ch_reference_index = Channel.fromPath(params.genomes[ params.genome ].reference_index, checkIfExists: true) + .collect() | EMIT_META_REF + ch_chrom_dir = Channel.fromPath("${params.genomes[ params.genome ].chromosomes_dir}", type: 'dir', checkIfExists: true) + ch_chrom_sizes = Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) + ch_gene_info = Channel.fromPath(params.genomes[ params.genome ].gene_info, checkIfExists: true) + ch_gsize = Channel.value(params.genomes[ params.genome ].effective_genome_size) + + } else if (params.genome_fasta && params.genes_gtf && params.blacklist) { + println "Building a reference from provided genome fasta, gtf, and blacklist files" + ch_fasta = file(params.genome_fasta, checkIfExists: true) + ch_blacklist_input = file(params.blacklist, checkIfExists: true) if (ch_blacklist_input.endsWith('.bed')) { BEDTOOLS_GETFASTA(ch_blacklist_input, ch_fasta) ch_blacklist_fasta = BEDTOOLS_GETFASTA.out.fasta } else { ch_blacklist_fasta = ch_blacklist_input } - ch_blacklist_index = BWA_INDEX_BL([['id': 'blacklist'], ch_blacklist_fasta]).index - ch_reference_index = BWA_INDEX_REF([['id': 'genome'], ch_fasta]).index + ch_blacklist_index = BWA_INDEX_BL([[id: 'blacklist'], ch_blacklist_fasta]).index + ch_reference_index = BWA_INDEX_REF([[id: 'genome'], ch_fasta]).index KHMER_UNIQUEKMERS(ch_fasta, params.read_length) ch_gsize = KHMER_UNIQUEKMERS.out.kmers.map { it.text.trim() } - ch_gene_info = GTF2BED ( file(params.genes_gtf) ).bed + ch_gene_info = GTF2BED ( file(params.genes_gtf, checkIfExists: true) ).bed SPLIT_REF_CHROMS(ch_fasta) ch_chrom_sizes = SPLIT_REF_CHROMS.out.chrom_sizes ch_chrom_dir = SPLIT_REF_CHROMS.out.chrom_dir - // save reference files - if (params.save_reference) { - //file("${params.outdir}/genome/").mkdirs() - WRITE_GENOME_CONFIG( - ch_reference_index, - ch_blacklist_index, - ch_chrom_sizes, - ch_chrom_dir, - ch_gene_info, - ch_gsize - ) - - } + WRITE_GENOME_CONFIG( + ch_reference_index, + ch_blacklist_index, + ch_chrom_sizes, + ch_chrom_dir, + ch_gene_info, + ch_gsize + ) + println "Saving custom genome config in ${params.outdir}/genome" } else { - Channel.fromPath(params.genomes[ params.genome ].blacklist_index, checkIfExists: true) - .collect() - .set{ ch_blacklist_index } - Channel.fromPath(params.genomes[ params.genome ].reference_index, checkIfExists: true) - .collect() - .set{ ch_reference_index } - Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) - .set{ ch_chrom_sizes } - Channel.fromPath("${params.genomes[ params.genome ].chromosomes_dir}", type: 'dir', checkIfExists: true) - .set{ ch_chrom_dir } - Channel.fromPath(params.genomes[ params.genome ].gene_info, - checkIfExists: true) - .set{ ch_gene_info } - Channel.value(params.genomes[ params.genome ].effective_genome_size) - .set{ ch_gsize} + error "Either specify a genome in `conf/genomes.conf`, or specify a genome fasta, gtf, and blacklist file to build a custom reference." } emit: From 4718afb0c89158d73889d2866dd756466d4a2b53 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 15:08:22 -0400 Subject: [PATCH 063/100] feat: separate entry point to prepare the reference genome --- main.nf | 4 ++++ 1 file changed, 4 insertions(+) diff --git a/main.nf b/main.nf index bcf6b8c3..681c11bf 100644 --- a/main.nf +++ b/main.nf @@ -37,6 +37,10 @@ include { PPQT_PROCESS MULTIQC } from "./modules/local/qc.nf" include { NORMALIZE_INPUT } from "./modules/local/deeptools.nf" +workflow MAKE_REFERENCE { + PREPARE_GENOME() +} + // MAIN WORKFLOW workflow { INPUT_CHECK(file(params.input), params.seq_center) From fd9cae2594245daf0e6faf1593752f6e6cf946f4 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 15:08:49 -0400 Subject: [PATCH 064/100] chore: reorder for readability --- conf/genomes.config | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/conf/genomes.config b/conf/genomes.config index 28b6ce46..7be7eb38 100644 --- a/conf/genomes.config +++ b/conf/genomes.config @@ -3,18 +3,18 @@ params { 'hg38' { blacklist_index = "${params.index_dir}/hg38_basic/indexes/blacklist/*" reference_index = "${params.index_dir}/hg38_basic/indexes/hg38*" - effective_genome_size = 2700000000 + chromosomes_dir = "${params.index_dir}/hg38_basic/Chromsomes/" chrom_sizes = "${params.index_dir}/hg38_basic/indexes/hg38.fa.sizes" gene_info = "${params.index_dir}/hg38_basic/geneinfo.bed" - chromosomes_dir = "${params.index_dir}/hg38_basic/Chromsomes/" + effective_genome_size = 2700000000 } 'mm10' { blacklist_index = "${params.index_dir}/mm10_basic/indexes/mm10.blacklist.chrM.chr_rDNA.*" reference_index = "${params.index_dir}/mm10_basic/indexes/mm10*" - effective_genome_size = 2400000000 + chromosomes_dir = "${params.index_dir}/mm10_basic/Chromsomes/" chrom_sizes = "${params.index_dir}/mm10_basic/indexes/mm10.fa.sizes" gene_info = "${params.index_dir}/mm10_basic/geneinfo.bed" - chromosomes_dir = "${params.index_dir}/mm10_basic/Chromsomes/" + effective_genome_size = 2400000000 } } } From f0f96565208a0564de57bc0bbf4a9b610ff36e38 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 4 Oct 2023 15:09:02 -0400 Subject: [PATCH 065/100] chore: reorder for readability --- conf/modules.config | 6 ------ conf/test.config | 2 +- conf/test_paired.config | 4 ++-- 3 files changed, 3 insertions(+), 9 deletions(-) diff --git a/conf/modules.config b/conf/modules.config index 5723e2bc..2496842b 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -24,10 +24,4 @@ process { '-b file:/opt2/TruSeq_and_nextera_adapters.consolidated.fa' ].join(' ').trim() } - withName: 'WRITE_GENOME_CONFIG' { - publishDir = [ - path: { "${params.outdir}/genome" }, - mode: "copy" - ] - } } diff --git a/conf/test.config b/conf/test.config index 755b1aba..8107dd20 100644 --- a/conf/test.config +++ b/conf/test.config @@ -5,13 +5,13 @@ params { outdir = 'results/test' input = 'assets/samplesheet_test.csv' // adapted from https://github.com/nf-core/test-datasets/blob/chipseq/samplesheet/v2.0/samplesheet_test.csv + genome = 'custom_genome' read_length = 50 // Genome references genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' - genome = 'sacCer3' save_reference = true // https://github.com/nf-core/chipseq/blob/51eba00b32885c4d0bec60db3cb0a45eb61e34c5/subworkflows/local/prepare_genome.nf#L47-L51 max_cpus = 32 // for interactive node on biowulf diff --git a/conf/test_paired.config b/conf/test_paired.config index 585a059a..bf7f0372 100644 --- a/conf/test_paired.config +++ b/conf/test_paired.config @@ -3,9 +3,9 @@ params { config_profile_description = 'Minimal test dataset to check paired & single end handling' outdir = 'results/test_paired' - input = 'assets/samplesheet_test_paired.csv' // adapted from 'https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/samplesheet/v2.0/samplesheet_test.csv' - genome = 'sacCer3' + + genome = 'custom_genome' read_length = 50 // Genome references From c4d238b57e3cd2875690849991ec52eab24e0d58 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Thu, 5 Oct 2023 23:14:45 -0400 Subject: [PATCH 066/100] chore: update changelog --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 95a34d86..453ba1ea 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -8,6 +8,7 @@ - Fraction in Peaks (FRiP) (#89) - Jaccard index (#92) - Histogram of peak widths (#92) +- Added an option to use a custom reference from a genome fasta, gtf, and blacklist file. ### Bug fixes From 06bef80866bf403adf61a43bfc2c9028cc20cae2 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Thu, 5 Oct 2023 23:48:27 -0400 Subject: [PATCH 067/100] fix: must match exactly one bwa index --- conf/genomes.config | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/conf/genomes.config b/conf/genomes.config index 7be7eb38..ae791b29 100644 --- a/conf/genomes.config +++ b/conf/genomes.config @@ -1,16 +1,16 @@ params { genomes { 'hg38' { - blacklist_index = "${params.index_dir}/hg38_basic/indexes/blacklist/*" - reference_index = "${params.index_dir}/hg38_basic/indexes/hg38*" + blacklist_index = "${params.index_dir}/hg38_basic/indexes/blacklist/hg38.blacklist_v3.chrM.chr_rDNA.*" + reference_index = "${params.index_dir}/hg38_basic/bwa_index/hg38*" chromosomes_dir = "${params.index_dir}/hg38_basic/Chromsomes/" chrom_sizes = "${params.index_dir}/hg38_basic/indexes/hg38.fa.sizes" gene_info = "${params.index_dir}/hg38_basic/geneinfo.bed" effective_genome_size = 2700000000 } 'mm10' { - blacklist_index = "${params.index_dir}/mm10_basic/indexes/mm10.blacklist.chrM.chr_rDNA.*" - reference_index = "${params.index_dir}/mm10_basic/indexes/mm10*" + blacklist_index = "${params.index_dir}/mm10_basic/indexes/blacklist/mm10.blacklist.chrM.chr_rDNA.*" + reference_index = "${params.index_dir}/mm10_basic/indexes/reference/mm10*" chromosomes_dir = "${params.index_dir}/mm10_basic/Chromsomes/" chrom_sizes = "${params.index_dir}/mm10_basic/indexes/mm10.fa.sizes" gene_info = "${params.index_dir}/mm10_basic/geneinfo.bed" From b9698903cf4386f36d255dbd9794df54847a1704 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 6 Oct 2023 11:30:34 -0400 Subject: [PATCH 068/100] fix: filter out negative gem peaks --- modules/local/peaks.nf | 51 +++++++++++++++++++++++++++++++++++++ subworkflows/local/peaks.nf | 8 ++++-- 2 files changed, 57 insertions(+), 2 deletions(-) diff --git a/modules/local/peaks.nf b/modules/local/peaks.nf index 16c7306d..f803bc52 100644 --- a/modules/local/peaks.nf +++ b/modules/local/peaks.nf @@ -255,6 +255,57 @@ process GEM { """ } +process MAKE_CHROM_FILTER { + // to filter out negative peaks from GEM + container "${params.containers.base}" + + input: + path(chrom_sizes) + + output: + path("${chrom_sizes}.filter.txt"), emit: txt + + script: + """ + #!/usr/bin/env python + with open("${chrom_sizes}", "r") as infile: + with open("${chrom_sizes}.filter.txt", "w") as outfile: + for line in infile: + chrom_name, chrom_len = line.strip().split() + outfile.write(f"{chrom_name}\\t1\\t1\\n") + outfile.write(f"{chrom_name}\\t{chrom_len}\\t{chrom_len}\\n") + """ + + stub: + """ + touch ${chrom_sizes}.filter.txt + """ + +} + +process FILTER_GEM { + tag { meta.id } + + container "${params.containers.base}" + + input: + tuple val(meta), path(peak), val(tool) + path(chrom_filter) + + output: + tuple val(meta), path("${peak}.filtered"), val(tool), emit: peak + + script: + """ + bedtools intersect -v -a ${peak} -b ${chrom_filter} > ${peak}.filtered + """ + stub: + """ + touch ${peak}.filtered + """ + +} + process FRACTION_IN_PEAKS { tag { meta.id } label 'peaks' diff --git a/subworkflows/local/peaks.nf b/subworkflows/local/peaks.nf index ab82e42e..67c585c1 100644 --- a/subworkflows/local/peaks.nf +++ b/subworkflows/local/peaks.nf @@ -5,6 +5,8 @@ include { CALC_GENOME_FRAC SICER CONVERT_SICER GEM + MAKE_CHROM_FILTER + FILTER_GEM FRACTION_IN_PEAKS CONCAT_FRIPS PLOT_FRIP @@ -70,7 +72,7 @@ workflow CALL_PEAKS { .set { ch_sicer } - // create gem channel with [ meta, chip_tag, input_tag, format, read_dists, chrom_sizes, chrom_dir ] + // create gem channel with [ meta, chip_tag, input_tag, format, read_dists, chrom_sizes, chrom_dir, effective_genome_sizes ] ch_tagalign .combine(Channel.fromPath(params.gem.read_dists, checkIfExists: true)) .combine(chrom_sizes) @@ -93,7 +95,9 @@ workflow CALL_PEAKS { } if (params.run.gem) { ch_gem | GEM - ch_peaks = ch_peaks.mix(GEM.out.peak) + MAKE_CHROM_FILTER(chrom_sizes) + FILTER_GEM(GEM.out.peak, MAKE_CHROM_FILTER.out.txt) + ch_peaks = ch_peaks.mix(FILTER_GEM.out.peak) } // Create Channel with meta, deduped bam, peak file, peak-calling tool, and chrom sizes fasta From fa09eb97a1da2f2bd599bf8b067d562f759e8a23 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 6 Oct 2023 11:55:46 -0400 Subject: [PATCH 069/100] fix: save genome fasta too --- conf/test.config | 1 - modules/local/prepare_genome.nf | 4 +++- subworkflows/local/prepare_genome.nf | 1 + 3 files changed, 4 insertions(+), 2 deletions(-) diff --git a/conf/test.config b/conf/test.config index 8107dd20..3b9989f4 100644 --- a/conf/test.config +++ b/conf/test.config @@ -12,7 +12,6 @@ params { genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' - save_reference = true // https://github.com/nf-core/chipseq/blob/51eba00b32885c4d0bec60db3cb0a45eb61e34c5/subworkflows/local/prepare_genome.nf#L47-L51 max_cpus = 32 // for interactive node on biowulf max_memory = '120 GB' // for interactive node on biowulf diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index 87c30022..1a3cb92a 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -24,6 +24,7 @@ process GTF2BED { script: """ cat ${gtf} | gtf2bed > ${gtf.baseName}.bed + """ stub: """ @@ -63,6 +64,7 @@ process WRITE_GENOME_CONFIG { mode: "copy" ] input: + path(fasta) tuple val(meta_ref), path(reference_index) tuple val(meta_bl), path(blacklist_index) path(chrom_sizes) @@ -88,7 +90,7 @@ process WRITE_GENOME_CONFIG { for file in filelist.split(): shutil.copy(file, dirpath) shutil.copytree("${chrom_dir}", '${genome_name}/chroms/') - for file in ("${chrom_sizes}", "${gene_info}"): + for file in ("${fasta}", "${chrom_sizes}", "${gene_info}"): shutil.copy(file, "${genome_name}/") genome = dict(reference_index = '"\${params.index_dir}/${genome_name}/reference/*"', diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index e13771d5..566ae674 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -42,6 +42,7 @@ workflow PREPARE_GENOME { ch_chrom_dir = SPLIT_REF_CHROMS.out.chrom_dir WRITE_GENOME_CONFIG( + ch_fasta, ch_reference_index, ch_blacklist_index, ch_chrom_sizes, From 1719471824e83c2496588b77f57b1047ab04013f Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 6 Oct 2023 16:15:26 -0400 Subject: [PATCH 070/100] fix: sicer genome --- conf/ci_stub.config | 3 +++ conf/full_mm10.config | 3 +++ conf/test_mm10.config | 3 +++ nextflow.config | 2 +- 4 files changed, 10 insertions(+), 1 deletion(-) diff --git a/conf/ci_stub.config b/conf/ci_stub.config index 078c0b7e..73f8c37b 100644 --- a/conf/ci_stub.config +++ b/conf/ci_stub.config @@ -28,6 +28,9 @@ params { chromosomes_dir = "tests/data/chroms/" } } + sicer { + species = "sacCer1" // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py + } } process { diff --git a/conf/full_mm10.config b/conf/full_mm10.config index ae4a6696..8e77b7d9 100644 --- a/conf/full_mm10.config +++ b/conf/full_mm10.config @@ -5,4 +5,7 @@ params { genome = 'mm10' outdir = "results/full_mm10" input = "assets/samplesheet_full_mm10.csv" + sicer { + species = "mm10" // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py + } } diff --git a/conf/test_mm10.config b/conf/test_mm10.config index 0b40733e..5e7a87fa 100644 --- a/conf/test_mm10.config +++ b/conf/test_mm10.config @@ -6,4 +6,7 @@ params { outdir = "results/test_mm10" input = "assets/samplesheet_test_mm10.csv" read_length = 50 + sicer { + species = "mm10" // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py + } } diff --git a/nextflow.config b/nextflow.config index 6a48d0c4..b0133850 100644 --- a/nextflow.config +++ b/nextflow.config @@ -62,7 +62,7 @@ params { } } sicer { - species = "sacCer1" // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py + species = null // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py } run { // some steps can be turned on/off for debugging purposes From d727cab79c9e6ba9ebdda11f6494265a41735d0a Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Fri, 6 Oct 2023 16:40:02 -0400 Subject: [PATCH 071/100] fix: keep log dir --- log/.gitkeep | 0 1 file changed, 0 insertions(+), 0 deletions(-) create mode 100644 log/.gitkeep diff --git a/log/.gitkeep b/log/.gitkeep new file mode 100644 index 00000000..e69de29b From 232c79a3c9abb259aba07ffabd6faa85c4765376 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 11:38:39 -0400 Subject: [PATCH 072/100] chore: set default sicer species to same as genome --- nextflow.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index b0133850..ef290f8a 100644 --- a/nextflow.config +++ b/nextflow.config @@ -62,7 +62,7 @@ params { } } sicer { - species = null // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py + species = "${params.genome}" // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py } run { // some steps can be turned on/off for debugging purposes From 32371a6b3c21dd5fb489db4a19cc1f6f649132d0 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 11:53:32 -0400 Subject: [PATCH 073/100] fix: filter gem peaks -- run on all peaks --- modules/local/peaks.nf | 3 +-- subworkflows/local/peaks.nf | 6 ++++-- 2 files changed, 5 insertions(+), 4 deletions(-) diff --git a/modules/local/peaks.nf b/modules/local/peaks.nf index f803bc52..5e17d5b5 100644 --- a/modules/local/peaks.nf +++ b/modules/local/peaks.nf @@ -289,8 +289,7 @@ process FILTER_GEM { container "${params.containers.base}" input: - tuple val(meta), path(peak), val(tool) - path(chrom_filter) + tuple val(meta), path(peak), val(tool), path(chrom_filter) output: tuple val(meta), path("${peak}.filtered"), val(tool), emit: peak diff --git a/subworkflows/local/peaks.nf b/subworkflows/local/peaks.nf index 67c585c1..db33244b 100644 --- a/subworkflows/local/peaks.nf +++ b/subworkflows/local/peaks.nf @@ -95,8 +95,10 @@ workflow CALL_PEAKS { } if (params.run.gem) { ch_gem | GEM - MAKE_CHROM_FILTER(chrom_sizes) - FILTER_GEM(GEM.out.peak, MAKE_CHROM_FILTER.out.txt) + GEM.out.peak + .combine( + MAKE_CHROM_FILTER(chrom_sizes).txt + ) | FILTER_GEM ch_peaks = ch_peaks.mix(FILTER_GEM.out.peak) } From 9e066cb8e2c00c1d978d711312a8c563d632f67d Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 12:54:44 -0400 Subject: [PATCH 074/100] feat: rename yeast contigs from ensembl to ucsc --- conf/test.config | 1 + conf/test_paired.config | 5 ++- modules/local/prepare_genome.nf | 65 ++++++++++++++++++++++++++++ nextflow.config | 2 +- subworkflows/local/prepare_genome.nf | 47 ++++++++++++++------ 5 files changed, 104 insertions(+), 16 deletions(-) diff --git a/conf/test.config b/conf/test.config index 3b9989f4..c9332b75 100644 --- a/conf/test.config +++ b/conf/test.config @@ -12,6 +12,7 @@ params { genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' + rename_contigs = 'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' max_cpus = 32 // for interactive node on biowulf max_memory = '120 GB' // for interactive node on biowulf diff --git a/conf/test_paired.config b/conf/test_paired.config index bf7f0372..8a5219e3 100644 --- a/conf/test_paired.config +++ b/conf/test_paired.config @@ -12,7 +12,8 @@ params { genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' - save_reference = true + rename_contigs = 'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' + max_cpus = 32 // for interactive node on biowulf max_memory = '120 GB' // for interactive node on biowulf @@ -22,7 +23,7 @@ params { qc = false normalize_input = false call_peaks = true - gem = false + gem = true sicer = false macs_broad = true macs_narrow = true diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index 1a3cb92a..59a1f83c 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -55,6 +55,71 @@ process SPLIT_REF_CHROMS { """ } +process RENAME_FASTA_CONTIGS { + """ + Convert ensembl to UCSC contig names in a fasta file + """ + tag { fasta } + + container "${params.containers.base}" + + input: + path(fasta) + path(map) + + output: + tuple val(fasta.baseName), path("*_renamed*"), emit: fasta + + script: + def renamed_fasta= "${fasta.getSimpleName()}_renamed.${fasta.getExtension()}" + """ + #!/usr/bin/env python + + with open("${map}", "r") as mapfile: + contig_map = {line.strip().split()[0]: line.strip().split()[1] for line in mapfile} + + with open("${fasta}", "r") as infile: + with open("${renamed_fasta}", "w") as outfile: + for line in infile: + if line.startswith(">"): + old_contig = line.strip(">").strip() + contig = contig_map[contig] if old_contig in contig_map.keys() else old_contig + outfile.write(f">{config}\n") + else: + outfile.write(line) + + """ +} + +process RENAME_DELIM_CONTIGS { + """ + Convert ensembl to UCSC contig names in a delimited file (e.g. GTF, BED) + using cvbio https://github.com/clintval/cvbio#updatecontignames + """ + tag { delim } + + container "nciccbr/common_cvbio_3.0.0" + + input: + path(delim) + path(map) + + output: + tuple val(delim.baseName), path("*_renamed*"), emit: delim + + script: + def renamed_delim = "${delim.getSimpleName()}_renamed.${delim.getExtension()}" + """ + cvbio UpdateContigNames \\ + -i ${delim} \\ + -o ${renamed_delim} \\ + -m ${map} \\ + --comment-chars '#' \\ + --columns 0 \\ + --skip-missing false + """ +} + process WRITE_GENOME_CONFIG { label 'process_single' container "${params.containers.base}" diff --git a/nextflow.config b/nextflow.config index ef290f8a..0416c5f4 100644 --- a/nextflow.config +++ b/nextflow.config @@ -10,7 +10,7 @@ params { genome_fasta = null genes_gtf = null blacklist = null - save_reference = false + rename_contigs = null // Boilerplate options outdir = 'results' diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index 566ae674..869b6246 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -2,10 +2,13 @@ include { BWA_INDEX as BWA_INDEX_BL BWA_INDEX as BWA_INDEX_REF } from "../../modules/CCBR/bwa/index" include { KHMER_UNIQUEKMERS } from '../../modules/nf-core/khmer/uniquekmers' include { BEDTOOLS_GETFASTA } from '../../modules/nf-core/bedtools/getfasta/main' -include { SPLIT_REF_CHROMS } from '../../modules/local/prepare_genome.nf' -include { GTF2BED } from '../../modules/local/prepare_genome.nf' -include { WRITE_GENOME_CONFIG } from "../../modules/local/prepare_genome.nf" -include { EMIT_META as EMIT_META_BL +include { SPLIT_REF_CHROMS + RENAME_FASTA_CONTIGS as RENAME_FASTA_CONTIGS_REF + RENAME_FASTA_CONTIGS as RENAME_FASTA_CONTIGS_BL + RENAME_DELIM_CONTIGS + GTF2BED + WRITE_GENOME_CONFIG + EMIT_META as EMIT_META_BL EMIT_META as EMIT_META_REF } from "../../modules/local/prepare_genome.nf" workflow PREPARE_GENOME { @@ -24,19 +27,37 @@ workflow PREPARE_GENOME { } else if (params.genome_fasta && params.genes_gtf && params.blacklist) { println "Building a reference from provided genome fasta, gtf, and blacklist files" - ch_fasta = file(params.genome_fasta, checkIfExists: true) - ch_blacklist_input = file(params.blacklist, checkIfExists: true) - if (ch_blacklist_input.endsWith('.bed')) { - BEDTOOLS_GETFASTA(ch_blacklist_input, ch_fasta) - ch_blacklist_fasta = BEDTOOLS_GETFASTA.out.fasta + ch_fasta_input = file(params.genome_fasta, checkIfExists: true) + ch_gtf_input = file(params.genes_gtf, checkIfExists: true) + + // blacklist bed to fasta + ch_blacklist = file(params.blacklist, checkIfExists: true) + if (ch_blacklist.endsWith('.bed')) { + BEDTOOLS_GETFASTA(ch_blacklist, ch_fasta) + ch_blacklist_input = BEDTOOLS_GETFASTA.out.fasta } else { - ch_blacklist_fasta = ch_blacklist_input + ch_blacklist_input = ch_blacklist } - ch_blacklist_index = BWA_INDEX_BL([[id: 'blacklist'], ch_blacklist_fasta]).index - ch_reference_index = BWA_INDEX_REF([[id: 'genome'], ch_fasta]).index + + // rename contigs from ensembl to UCSC if needed + if (params.rename_contigs) { + contig_map = file(params.rename_contigs, checkIfExists: true) + println contig_map + ch_fasta = RENAME_FASTA_CONTIGS_REF(ch_fasta_input, contig_map).fasta + println ch_fasta + ch_blacklist_fasta = RENAME_FASTA_CONTIGS_BL(ch_blacklist_input, contig_map).fasta + ch_gtf = RENAME_DELIM_CONTIGS(ch_gtf_input, contig_map).delim + } else { + ch_fasta = [[id: 'genome'], ch_fasta_input] + ch_blacklist_fasta = [[id: 'blacklist'], ch_blacklist_input] + ch_gtf = ch_gtf_input + } + + ch_blacklist_index = BWA_INDEX_BL(ch_blacklist_fasta).index + ch_reference_index = BWA_INDEX_REF(ch_fasta).index KHMER_UNIQUEKMERS(ch_fasta, params.read_length) ch_gsize = KHMER_UNIQUEKMERS.out.kmers.map { it.text.trim() } - ch_gene_info = GTF2BED ( file(params.genes_gtf, checkIfExists: true) ).bed + ch_gene_info = GTF2BED ( ch_gtf ).bed SPLIT_REF_CHROMS(ch_fasta) ch_chrom_sizes = SPLIT_REF_CHROMS.out.chrom_sizes ch_chrom_dir = SPLIT_REF_CHROMS.out.chrom_dir From 076b75a8446f3236732a096c29b01a85435c1831 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 13:02:34 -0400 Subject: [PATCH 075/100] fix: stub for bwa mem --- modules.json | 2 +- modules/CCBR/bwa/mem/main.nf | 4 ++++ 2 files changed, 5 insertions(+), 1 deletion(-) diff --git a/modules.json b/modules.json index 14ccc61a..65ab6d7b 100644 --- a/modules.json +++ b/modules.json @@ -12,7 +12,7 @@ }, "bwa/mem": { "branch": "main", - "git_sha": "520860abefe5e45ac604e92051d2daebdae4aa81", + "git_sha": "29ac0b1a6c6b70f8fda38cd9c11c8a2c7f0ff6cd", "installed_by": ["modules"] }, "cutadapt": { diff --git a/modules/CCBR/bwa/mem/main.nf b/modules/CCBR/bwa/mem/main.nf index 4a8fd260..595e417a 100644 --- a/modules/CCBR/bwa/mem/main.nf +++ b/modules/CCBR/bwa/mem/main.nf @@ -39,4 +39,8 @@ process BWA_MEM { samtools: \$(echo \$(samtools --version 2>&1) | sed 's/^.*samtools //; s/Using.*\$//') END_VERSIONS """ + stub: + """ + touch ${meta.id}.bam versions.yml + """ } From cd044cc4cf620da63fe642ceec3d1de19711d248 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 13:07:35 -0400 Subject: [PATCH 076/100] fix: add/update stubs --- modules/local/align.nf | 2 +- modules/local/prepare_genome.nf | 16 ++++++++++++++++ 2 files changed, 17 insertions(+), 1 deletion(-) diff --git a/modules/local/align.nf b/modules/local/align.nf index d88484ce..c71683c9 100644 --- a/modules/local/align.nf +++ b/modules/local/align.nf @@ -58,7 +58,7 @@ process BAM_TO_FASTQ { } stub: """ - touch ${bam.baseName}.fastq + touch ${bam.baseName}.R1.fastq """ } diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index 59a1f83c..fb3d4b3f 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -89,6 +89,12 @@ process RENAME_FASTA_CONTIGS { outfile.write(line) """ + + stub: + def renamed_fasta= "${fasta.getSimpleName()}_renamed.${fasta.getExtension()}" + """ + touch ${renamed_fasta} + """ } process RENAME_DELIM_CONTIGS { @@ -118,6 +124,12 @@ process RENAME_DELIM_CONTIGS { --columns 0 \\ --skip-missing false """ + + stub: + def renamed_delim = "${delim.getSimpleName()}_renamed.${delim.getExtension()}" + """ + touch ${renamed_delim} + """ } process WRITE_GENOME_CONFIG { @@ -179,4 +191,8 @@ process WRITE_GENOME_CONFIG { "}\\n"] conf_file.writelines(tail) """ + + stub: + """ + """ } From 4c1694614fbf2984c8a65325b315d08bea65b846 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 13:16:56 -0400 Subject: [PATCH 077/100] fix: fastqc stub output to avoid file name collisions --- modules/local/qc.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/qc.nf b/modules/local/qc.nf index 7958656a..fe2a430c 100644 --- a/modules/local/qc.nf +++ b/modules/local/qc.nf @@ -23,7 +23,7 @@ process FASTQC { stub: """ - touch ${meta.id}_fastqc.html ${meta.id}_fastqc.zip + touch ${meta.id}_${fqtype}_fastqc.html ${meta.id}_${fqtype}_fastqc.zip """ } @@ -184,7 +184,7 @@ process PHANTOM_PEAKS { stub: """ - touch ${meta.id}.ppqt.pdf ${meta.id}.spp.out "${meta.id}.fraglen.txt" + touch ${meta.id}.ppqt.pdf ${meta.id}.spp.out ${meta.id}.fraglen.txt versions.yml """ } From a5761cf76d0c8b12e50d2d9a940275de437d2d6d Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 13:18:48 -0400 Subject: [PATCH 078/100] feat: use test_paired for ci_stub --- conf/ci_stub.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/ci_stub.config b/conf/ci_stub.config index 73f8c37b..f5ba0f94 100644 --- a/conf/ci_stub.config +++ b/conf/ci_stub.config @@ -3,7 +3,7 @@ params { config_profile_description = 'Minimal test dataset with blank references to run stubs with continuous integration' outdir = 'results/test' - input = 'assets/samplesheet_test.csv' // adapted from https://github.com/nf-core/test-datasets/blob/chipseq/samplesheet/v2.0/samplesheet_test.csv + input = 'assets/samplesheet_test_paired.csv' // adapted from https://github.com/nf-core/test-datasets/blob/chipseq/samplesheet/v2.0/samplesheet_test.csv genome = 'test' max_cpus = 2 // for GitHub Actions https://docs.github.com/en/actions/using-github-hosted-runners/about-github-hosted-runners#supported-runners-and-hardware-resources From 19322614ba236f6d67c70b6f29d1d9e47700fa28 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 14:10:37 -0400 Subject: [PATCH 079/100] fix: meta of fasta files --- modules/local/prepare_genome.nf | 12 ++------ subworkflows/local/prepare_genome.nf | 45 +++++++++++++++------------- 2 files changed, 27 insertions(+), 30 deletions(-) diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index fb3d4b3f..faf98df0 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -1,13 +1,3 @@ -process EMIT_META { - input: - path(file) - output: - tuple val(file.baseName), path(file) - script: - """ - echo $file - """ -} process GTF2BED { tag { gtf } @@ -194,5 +184,7 @@ process WRITE_GENOME_CONFIG { stub: """ + mkdir custom_genome/ + touch custom_genome.config custom_genome/genome.fa """ } diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index 869b6246..84c1f834 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -7,9 +7,7 @@ include { SPLIT_REF_CHROMS RENAME_FASTA_CONTIGS as RENAME_FASTA_CONTIGS_BL RENAME_DELIM_CONTIGS GTF2BED - WRITE_GENOME_CONFIG - EMIT_META as EMIT_META_BL - EMIT_META as EMIT_META_REF } from "../../modules/local/prepare_genome.nf" + WRITE_GENOME_CONFIG } from "../../modules/local/prepare_genome.nf" workflow PREPARE_GENOME { main: @@ -17,9 +15,15 @@ workflow PREPARE_GENOME { println "Using ${params.genome} as the reference" ch_blacklist_index = Channel.fromPath(params.genomes[ params.genome ].blacklist_index, checkIfExists: true) - .collect() | EMIT_META_BL + .collect() + .map{ file -> + [file.baseName, file] + } ch_reference_index = Channel.fromPath(params.genomes[ params.genome ].reference_index, checkIfExists: true) - .collect() | EMIT_META_REF + .collect() + .map{ file -> + [file.baseName, file] + } ch_chrom_dir = Channel.fromPath("${params.genomes[ params.genome ].chromosomes_dir}", type: 'dir', checkIfExists: true) ch_chrom_sizes = Channel.fromPath(params.genomes[ params.genome ].chrom_sizes, checkIfExists: true) ch_gene_info = Channel.fromPath(params.genomes[ params.genome ].gene_info, checkIfExists: true) @@ -27,34 +31,35 @@ workflow PREPARE_GENOME { } else if (params.genome_fasta && params.genes_gtf && params.blacklist) { println "Building a reference from provided genome fasta, gtf, and blacklist files" - ch_fasta_input = file(params.genome_fasta, checkIfExists: true) - ch_gtf_input = file(params.genes_gtf, checkIfExists: true) + fasta_file = Channel.fromPath(params.genome_fasta, checkIfExists: true) + gtf_file = file(params.genes_gtf, checkIfExists: true) // blacklist bed to fasta - ch_blacklist = file(params.blacklist, checkIfExists: true) - if (ch_blacklist.endsWith('.bed')) { - BEDTOOLS_GETFASTA(ch_blacklist, ch_fasta) + blacklist_file = file(params.blacklist, checkIfExists: true) + if (blacklist_file.endsWith('.bed')) { + BEDTOOLS_GETFASTA(blacklist_file, fasta_file) ch_blacklist_input = BEDTOOLS_GETFASTA.out.fasta } else { - ch_blacklist_input = ch_blacklist + ch_blacklist_input = Channel.fromPath(blacklist_file) } // rename contigs from ensembl to UCSC if needed if (params.rename_contigs) { contig_map = file(params.rename_contigs, checkIfExists: true) - println contig_map - ch_fasta = RENAME_FASTA_CONTIGS_REF(ch_fasta_input, contig_map).fasta - println ch_fasta + ch_fasta = RENAME_FASTA_CONTIGS_REF(fasta_file, contig_map).fasta ch_blacklist_fasta = RENAME_FASTA_CONTIGS_BL(ch_blacklist_input, contig_map).fasta - ch_gtf = RENAME_DELIM_CONTIGS(ch_gtf_input, contig_map).delim + ch_gtf = RENAME_DELIM_CONTIGS(gtf_file, contig_map).delim } else { - ch_fasta = [[id: 'genome'], ch_fasta_input] - ch_blacklist_fasta = [[id: 'blacklist'], ch_blacklist_input] - ch_gtf = ch_gtf_input + ch_fasta = fasta_file + ch_blacklist_fasta = ch_blacklist_input + ch_gtf = gtf_file } - ch_blacklist_index = BWA_INDEX_BL(ch_blacklist_fasta).index - ch_reference_index = BWA_INDEX_REF(ch_fasta).index + blacklist_meta = ch_blacklist_fasta.map{ it -> [it.baseName, it]} + fasta_meta = ch_fasta.map{ it -> [it.baseName, it]} + + ch_blacklist_index = BWA_INDEX_BL(blacklist_meta).index + ch_reference_index = BWA_INDEX_REF(fasta_meta).index KHMER_UNIQUEKMERS(ch_fasta, params.read_length) ch_gsize = KHMER_UNIQUEKMERS.out.kmers.map { it.text.trim() } ch_gene_info = GTF2BED ( ch_gtf ).bed From ef815b5569231d76457644afebd764d5c7f6fe40 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 14:29:39 -0400 Subject: [PATCH 080/100] feat: optionally skip fastq-screen for CI it needs absolute paths in the config file --- assets/fastq_screen_ci.conf | 41 ++++++++++++++++++ conf/ci_stub.config | 4 +- conf/ci_test.config | 35 +++++++++++++++ nextflow.config | 3 ++ subworkflows/local/qc.nf | 35 +++++---------- .../fastq_screen_db/rRNA/rRNA.human.2.bt2 | Bin 0 -> 10752 bytes .../fastq_screen_db/rRNA/rRNA.human.3.bt2 | Bin 0 -> 107 bytes .../fastq_screen_db/rRNA/rRNA.human.4.bt2 | Bin 0 -> 10747 bytes 8 files changed, 92 insertions(+), 26 deletions(-) create mode 100755 assets/fastq_screen_ci.conf create mode 100644 conf/ci_test.config create mode 100644 tests/data/fastq_screen_db/rRNA/rRNA.human.2.bt2 create mode 100644 tests/data/fastq_screen_db/rRNA/rRNA.human.3.bt2 create mode 100644 tests/data/fastq_screen_db/rRNA/rRNA.human.4.bt2 diff --git a/assets/fastq_screen_ci.conf b/assets/fastq_screen_ci.conf new file mode 100755 index 00000000..afdfb654 --- /dev/null +++ b/assets/fastq_screen_ci.conf @@ -0,0 +1,41 @@ +# This is a configuration file for fastq_screen + +############## +## Databases # +############## +## This section allows you to configure multiple databases +## to search against in your screen. For each database +## you need to provide a database name (which can't contain +## spaces) and the location of the bowtie indices which +## you created for that database. +## +## The entries shown below are only suggested examples, you +## can add as many DATABASE sections as required, and you +## can comment out or remove as many of the existing entries +## as desired. +## +## Either the original bowtie or bowtie2 may be used for the +## mapping. Specify the aligner to use with the command line +## flag --aligner with arguments 'bowtie' or +## 'bowtie2' (default). +## +## The configuration file may list paths to both bowtie and +## bowtie2 indices. FastQ Screen automatically detects whether +## a specified index is compatible with bowtie or bowtie2. +## +## Although the configuration file may list paths to both +## bowtie and bowtie2 indices, only one aligner will be used +## for the mapping, as specified by the --aligner flag. +## +## The path to the index files SHOULD INCLUDE THE BASENAME of +## the index, e.g: +## /data/public/Genomes/Human_Bowtie/GRCh37/Homo_sapiens.GRCh37 +## Thus, the indices (Homo_sapiens.GRCh37.1.bt2, Homo_sapiens.GRCh37.2.bt2, etc.) +## are found in a folder named 'GRCh37'. +## +## If the bowtie AND bowtie2 indices of a given genome reside in the SAME FOLDER, +## a SINGLE path may be provided to BOTH sets of indices. +## +## Human - sequences available from +## ftp://ftp.ensembl.org/pub/current/fasta/homo_sapiens/dna/ +DATABASE rRNA fastq_screen_db/rRNA/rRNA diff --git a/conf/ci_stub.config b/conf/ci_stub.config index f5ba0f94..795e9db5 100644 --- a/conf/ci_stub.config +++ b/conf/ci_stub.config @@ -15,8 +15,8 @@ params { // CCBR shared resource paths index_dir = "tests/data" fastq_screen { - conf = "assets/fastq_screen_biowulf.conf" - db_dir = "tests/data/" + conf = "assets/fastq_screen_ci.conf" + db_dir = "tests/data/fastq_screen_db" } genomes { 'test' { // blank files for testing stubs on GitHub Actions diff --git a/conf/ci_test.config b/conf/ci_test.config new file mode 100644 index 00000000..fd3d89b4 --- /dev/null +++ b/conf/ci_test.config @@ -0,0 +1,35 @@ +params { + config_profile_name = 'Test single-end stubs' + config_profile_description = 'Minimal test dataset with blank references to run stubs with continuous integration' + + outdir = 'results/test' + input = 'assets/samplesheet_test_paired.csv' // adapted from https://github.com/nf-core/test-datasets/blob/chipseq/samplesheet/v2.0/samplesheet_test.csv + + genome = 'custom_genome' + read_length = 50 + + // Genome references + genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' + genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' + blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' + rename_contigs = null //'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' + + + max_cpus = 2 // for GitHub Actions https://docs.github.com/en/actions/using-github-hosted-runners/about-github-hosted-runners#supported-runners-and-hardware-resources + max_memory = '6.GB' + max_time = '6.h' + + publish_dir_mode = "symlink" + + // CCBR shared resource paths + index_dir = "tests/data" + fastq_screen = null + sicer { + species = "sacCer1" // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py + } +} + +process { + cpus = 1 + memory = '1 GB' +} diff --git a/nextflow.config b/nextflow.config index 0416c5f4..010c27a7 100644 --- a/nextflow.config +++ b/nextflow.config @@ -115,6 +115,9 @@ profiles { ci_stub { includeConfig "conf/ci_stub.config" } + ci_test { + includeConfig "conf/ci_test.config" + } } includeConfig 'conf/genomes.config' diff --git a/subworkflows/local/qc.nf b/subworkflows/local/qc.nf index 30523cf0..2a377e35 100644 --- a/subworkflows/local/qc.nf +++ b/subworkflows/local/qc.nf @@ -35,11 +35,14 @@ workflow QC { main: raw_fastqs.combine(Channel.value("raw")) | FASTQC_RAW trimmed_fastqs.combine(Channel.value("trimmed")) | FASTQC_TRIMMED - trimmed_fastqs - .combine(Channel.fromPath(params.fastq_screen.conf, checkIfExists: true)) - .combine(Channel.fromPath(params.fastq_screen.db_dir, - type: 'dir', checkIfExists: true)) | FASTQ_SCREEN - + ch_multiqc = Channel.empty() + if (params.fastq_screen) { + trimmed_fastqs + .combine(Channel.fromPath(params.fastq_screen.conf, checkIfExists: true)) + .combine(Channel.fromPath(params.fastq_screen.db_dir, + type: 'dir', checkIfExists: true)) | FASTQ_SCREEN + ch_multiqc = ch_multiqc.mix(FASTQ_SCREEN.out.screen) + } PRESEQ(aligned_bam) // when preseq fails, write NAs for the stats that are calculated from its log PRESEQ.out.log @@ -101,26 +104,10 @@ workflow QC { meta1.control == meta2.id ? 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Sat, 7 Oct 2023 14:43:15 -0400 Subject: [PATCH 081/100] fix: collect bwa index output channel --- subworkflows/local/prepare_genome.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index 84c1f834..b1787d73 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -58,8 +58,8 @@ workflow PREPARE_GENOME { blacklist_meta = ch_blacklist_fasta.map{ it -> [it.baseName, it]} fasta_meta = ch_fasta.map{ it -> [it.baseName, it]} - ch_blacklist_index = BWA_INDEX_BL(blacklist_meta).index - ch_reference_index = BWA_INDEX_REF(fasta_meta).index + ch_blacklist_index = BWA_INDEX_BL(blacklist_meta).index.collect() + ch_reference_index = BWA_INDEX_REF(fasta_meta).index.collect() KHMER_UNIQUEKMERS(ch_fasta, params.read_length) ch_gsize = KHMER_UNIQUEKMERS.out.kmers.map { it.text.trim() } ch_gene_info = GTF2BED ( ch_gtf ).bed From 2847a3a11d250956adb259661cce9f24d77f7c83 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 14:45:27 -0400 Subject: [PATCH 082/100] test: update configs --- conf/ci_test.config | 4 +++- conf/test.config | 4 ++-- 2 files changed, 5 insertions(+), 3 deletions(-) diff --git a/conf/ci_test.config b/conf/ci_test.config index fd3d89b4..13ae9620 100644 --- a/conf/ci_test.config +++ b/conf/ci_test.config @@ -12,7 +12,7 @@ params { genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' - rename_contigs = null //'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' + rename_contigs = 'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' max_cpus = 2 // for GitHub Actions https://docs.github.com/en/actions/using-github-hosted-runners/about-github-hosted-runners#supported-runners-and-hardware-resources @@ -27,6 +27,8 @@ params { sicer { species = "sacCer1" // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py } + run.sicer = false // TODO set to true after https://github.com/CCBR/CHAMPAGNE/issues/109 + run.gem = true // TODO set to true after fixing rename_contigs } process { diff --git a/conf/test.config b/conf/test.config index c9332b75..054efe04 100644 --- a/conf/test.config +++ b/conf/test.config @@ -12,7 +12,7 @@ params { genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' - rename_contigs = 'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' + //rename_contigs = 'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' max_cpus = 32 // for interactive node on biowulf max_memory = '120 GB' // for interactive node on biowulf @@ -23,7 +23,7 @@ params { normalize_input = false call_peaks = true gem = true - sicer = false + sicer = false // TODO set to true after https://github.com/CCBR/CHAMPAGNE/issues/109 macs_broad = true macs_narrow = true } From 3e0b1612e8c1b1e64ac81ea9143a40cdf099c312 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 14:45:48 -0400 Subject: [PATCH 083/100] ci: run real test data on github actions --- .github/workflows/build.yml | 5 ++++- 1 file changed, 4 insertions(+), 1 deletion(-) diff --git a/.github/workflows/build.yml b/.github/workflows/build.yml index 935c4335..207c4633 100644 --- a/.github/workflows/build.yml +++ b/.github/workflows/build.yml @@ -29,9 +29,12 @@ jobs: run: | python -m pip install --upgrade pip setuptools pip install .[dev,test] - - name: Test stub run + - name: Stub run run: | champagne run -profile ci_stub -stub + - name: Test run + run: | + champagne run -profile ci_test,docker - name: "Upload Artifact" uses: actions/upload-artifact@v3 with: From 729f14348c554905949eca3760b5ea561d7c4f67 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 14:56:26 -0400 Subject: [PATCH 084/100] ci: generate fake blacklist for CI head -n 94 /data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa > tests/data/test.blacklist --- conf/ci_test.config | 2 +- tests/data/test.blacklist | 94 +++++++++++++++++++++++++++++++++++++++ 2 files changed, 95 insertions(+), 1 deletion(-) diff --git a/conf/ci_test.config b/conf/ci_test.config index 13ae9620..835b2ea8 100644 --- a/conf/ci_test.config +++ b/conf/ci_test.config @@ -11,7 +11,7 @@ params { // Genome references genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' - blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' + blacklist = 'tests/data/test.blacklist' rename_contigs = 'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' diff --git a/tests/data/test.blacklist b/tests/data/test.blacklist index e69de29b..13ee18d2 100644 --- a/tests/data/test.blacklist +++ b/tests/data/test.blacklist @@ -0,0 +1,94 @@ +>chr1:628903-635104 +TCACAACTTACTCACCAGATCAGACTTTGAGCTCTCCTCCTGGCTTAGCCTGGATCGTTTGAAATGGTCATCCATCCTTT +GGCCCCAATACCTAAACTAAGGTCTATGAACAATAAGATGATTTTCTTCAGTGGGACTTTTTTGTTTAATATATTAGATT +TGACCTTCAGCAAGGTCAAAGGGAGTCCGAACTAGTCTCAGGCTTCAACATCGAATACGCCGCAGGCCCCTTCGCCCTAT +TCTTCATAGCCGAATACACAAACATTATTATAATAAACACCCTCACCACTACAATCTTCCTAGGAACAACATATAACGCA +CTCTCCCCTGAACTCTACACAACATATTTTGTCACCAAGACCCTACTTCTGACCTCCCTGTTCTTATGAATTCGAACAGC +ATACCCCCGATTCCGCTACGACCAACTCATACACCTCCTATGAAAAAACTTCCTACCACTCACCCTAGCATTACTTATAT +GATATGTCTCCATACCCATTACAATCTCCAGCATTCCCCCTCAAACCTAAGAAATATGTCTGATAAAAGAGTTACTTTGA +TAGAGTAAATAATAGGAGTTTAAATCCCCTTATTTCTAGGACTATGAGAATCGAACCCATCCCTGAGAATCCAAAATTCT +CCGTGCCACCTATCACACCCCATCCTAAAGTAAGGTCAGCTAAATAAGCTATCGGGCCCATACCCCGAAAATGTTGGTTA +TATCCTTCCCGTACTAATTAATCCCCTGGCCCAACCCGTCATCTACTCTACCATCTTTGCAGGCACACTCATCACAGCGC +TAAGCTCGCACTGATTTTTTACCTGAGTAGGCCTAGAAATAAACATGCTAGCTTTTATTCCAGTTCTAACCAAAAAAATA +AACCCTCGTTCCACAGAAGCTGCCATCAAGTATTTCCTCACGCAAGCAACCGCATCCATAATCCTTCTAATAGCTATCCT +CTTCAACAATATACTCTCCGGACAATGAACCATAACCAATACCACCAATCAATACTCATCATTAATAATCATAATGGCTA +TAGCAATAAAACTAGGAATAGCCCCCTTTCACTTCTGAGTCCCAGAGGTTACCCAAGGCACCCCTCTGACATCCGGCCTG +CTCCTTCTCACATGACAAAAACTAGCCCCCATCTCAATCATATACCAAATTTCTCCCTCATTAAACGTAAGCCTTCTCCT +CACTCTTTCAATCTTATCCATCATGGCAGGCAGTTGAGGTGGATTAAACCAAACCCAACTACGCAAAATCTTAGCATACT +CCTCAATTACCCACATAGGATGAATAACAGCAGTTCTACCGTACAACCCTAACATAACCATTCTTAATTTAACTATTTAT +ATTATCCTAACTACTACCGCATTCCTACTACTCAACTTAAACTCCAGCACCACAACCCTACTACTATCTCGCACCTGAAA +CAAGCTAACATGACTAACACCCTTAATTCCATCCACCCTCCTCTCCCTAGGAGGCCTGCCCCCGCTAACCGGCTTTTTGC +CCAAATGGGCCATTATCGAAGAATTCACAAAAAACAATAGCCTCATCATCCCCACCATCATAGCCATCATCACCCTCCTT +AACCTCTACTTCTACCTGCGCCTAATCTACTCCACCTCAATCACACTACTCCCTATATCTAACAACGTAAAAATAAAATG +ACAGTTTGAACACACAAAACCCACCCCATTCCTCCCCACACTCATCGCCCTTACCACACTGCTCCTACCTATCTCCCCTT +TTATGCTAATAATCTTATAGAAATTTAGGTTAAATACAGACCAAGAGCCTTCAAAGCCCTCAGTAAGTTGCAATACTTAA +TTTCTGCAACAGCTAAGGACTGCAAAACCCCACTCTGCATCAACTGAACGCAAATCAGCCACTTTAATTAAGCTAAGCCC +TTACTAGACCAATGGGACTTAAACCCACAAACACTTAGTTAACAGCTAAGCACCCTAATCAACTGGCTTCAATCTACTTC +TCCCGCCGCCGGGAAAAAAGGCGGGAGAAGCCCCGGCAGGTTTGAAGCTGCTTCTTCGAATTTGCAATTCAATATGAAAA +TCACCTCAGAGCTGGTAAAAAGAGGCTTAACCCCTGTCTTTAGATTTACAGTCCAATGCTTCACTCAGCCATTTTACCTC +ACCCCCACTGATGTTCGCCGACCGTTGACTATTCTCTACAAACCACAAAGACATTGGAACACTATACCTATTATTCGGCG +CATGAGCTGGAGTCCTAGGCACAGCTCTAAGCCTCCTTATTCGAGCCGAACTGGGCCAGCCAGGCAACCTTCTAGGTAAC +GACCACATCTACAACGTTATCGTCACAGCCCATGCATTTGTAATAATCTTCTTCATAGTAATACCCATCATAATCGGAGG +CTTTGGCAACTGACTAGTTCCCCTAATAATCGGTGCCCCCGATATGGCGTTTCCCCGCATAAACAACATAAGCTTCTGAC +TCTTACCCCCCTCTCTCCTACTCCTGCTTGCATCTGCTATAGTGGAGGCCGGCGCAGGAACAGGTTGAACAGTCTACCCT +CCCTTGGCAGGGAACTACTCCCACCCTGGAGCCTCCGTAGACCTAACCATCTTCTCCTTACACCTAGCAGGTATCTCCTC +TATCTTAGGAGCCATCAATTTCATCACAACAATTATTAATATAAAACCCCCTGCCATAACCCAATACCAAACGCCCCTTT +TCGTCTGATCCGTCCTAATCACAGCAGTCTTACTTCTCCTATCTCTCCCAGTCCTAGCCGCTGGCATCACTATACTACTA +ACAGACCGTAACCTCAACACCACCTTCTTCGACCCAGCCGGAGGAGGAGACCCCATTCTATACCAACACCTATTCTGATT +TTTCGGTCACCCTGAAGTTTATATTCTCATCCTACCAGGCTTCGGAATAATCTCCCATATTGTAACTTACTACTCCGGGA +AAAAAAGAACCATTTGGATACATAGGTATGGTCTGAGCTATGATATCAATTGGCTTCCTAGGGTTTATCGTGTGAGCACA +CCATATATTTACAGTAGGAATAGACGTAGACACACGAGCATATTTCACCTCCGCTACCATAATCATCGCTATCCCCACCG +GCGTCAAAGTATTTAGCTGACTCGCCACACTCCACGGAAGCAATATGAAATGATCTGCTGCAGTGCTCTGAGCCCTAGGA +TTTATTTTTCTTTTCACCGTAGGTGGCCTGACTGGCATTGTATTAGCAAACTCATCACTAGACATCGTACTACACGACAC +GTACTACGTTGTAGCCCACTTCCACTATGTCCTATCAATAGGAGCTGTATTTGCCATCATAGGAGGCTTCATTCACTGAT +TTCCCCTATTCTCAGGCTACACCCTAGACCAAACCTACGCCAAAATCCATTTCGCTATCATATTCATCGGCGTAAATCTA +ACTTTCTTCCCACAACACTTTCTCGGCCTATCCGGAATGCCCCGACGTTACTCGGACTATCCCGATGCATACACCACATG +AAATATCCTATCATCTGTAGGCTCATTCATTTCTCTAACAGCAGTAATATTAATAATTTTCATAATTTGAGAAGCCTTCG +CTTCGAAGCGAAAAGTCCTAATAGTAGAAGAACCCTCCATAAACCTGGAGTGACTATATGGATGCCCCCCACCCTACCAC +ACATTCGAAGAACCCGTATACATAAAATCTAGACAAAAAAGGAAGGAATCGAACCCCCCAAAGCTGGTTTCAAGCCAACC +CCATGGCCTCCATGACTTTTTCAAAAAGATATTAGAAAAACCATTTCATAACTTTGTCAAAGTTAAATTATAGGCTAAAT +CCTATATATCTTAATGGCACATGCAGCGCAAGTAGGTCTACAAGACGCTACTTCCCCTATCATAGAAGAGCTTATCATCT +TTCATGATCACGCCCTCATAATCATTTTCCTTATCTGCTTCCTAGTCCTGTACGCCCTTTTCCTAACACTCACAACAAAA +CTAACTAATACTAACATCTCAGACGCTCAGGAAATAGAAACCGTCTGAACTATCCTGCCCGCCATCATCCTAGTCCTTAT +CGCCCTCCCATCCCTACGCATCCTTTACATAACAGACGAGGTCAACGATCCCTCCTTTACCATCAAATCAATTGGCCATC +AATGGTACTGAACCTACGAATACACCGACTACGGCGGACTAATCTTCAACTCCTACATACTTCCCCCATTATTCCTAGAA +CCAGGCGACCTGCGACTCCTTGACGTTGACAATCGAGTAGTACTCCCGGTTGAAGCCCCCATTCGTATAATAATTACATC +ACAAGACGTCTTACACTCATGAGCTGTCCCCACATTAGGCTTAAAAACAGATGCAATTCCCGGACGTCTAAACCAAACCA +CTTTCACTGCTACACGACCAGGGGTATACTACGGCCAATGCTCTGAAATCTGTGGAGCAAACCAGTTTTATGCCCATCGT +CCTAGAATTAATTCCCCTAAAAATCTTTGAAATAGGGCCCGTATTTACCCTATAGCACCCCCTCTACCCCCTCTAGAGCC +CACTGTAAAGCTAACTTAGCATTAACCTTTTAAGTTAAAGATTAAGAGAACCAACACCTCTTTACAGTGAAATGCCCCAA +CTAAATACTACCGTATGACCCACCATAATTACCCCCATACTCCTTACACTATTCCTCATCACCCAACTAAAAATATTAAA +TACAAATTACCACCTACCTCCCTCACCAAAGCCCATAAAAATAAAAAACTATAACAAACCCTGAGAACCAAAATGAACGA +AAATCTGTTCACTTCATTCATTGCCCCCACAATCCTAGGCCTACCCGCCGCAGTACTGATCATTCTATTTCCCCCTCTAT +TGATCCCCACCTCCAAATATCTCATCAACAACCGACTAATTACCACCCAACAATGACTAATCCAACTAACCTCAAAACAA +ATGATAGCCATACACAACACTAAGGGACGAACCTGATCTCTTATACTAGTATCCTTAATCATTTTTATTGCCACAACTAA +CCTCCTCGGACTCCTGCCTCACTCATTTACACCAACCACCCAACTATCTATAAACCTAGCCATGGCCATCCCCTTATGAG +CGGGCGCAGTGATTATAGGCTTTCGCTCTAAGATTAAAAATGCCCTAGCCCACTTCTTACCACAAGGCACACCTACACCC +CTTATCCCTATACTAGTTATTATCGAAACCATCAGCCTACTCATTCAACCAATAGCCCTGGCCGTACGCCTAACCGCTAA +CATTACTGCAGGCCACCTACTCATGCACCTAATTGGAAGCGCCACACTAGCAATATCAACTATTAACCTTCCCTCTACAC +TTATCATCTTCACAATTCTAATTCTACTGACTATCCTAGAAATCGCTGTCGCCTTAATCCAAGCCTACGTTTTTACACTT +CTAGTAAGCCTCTACCTGCACGACAACACATAATGACCCACCAATCACATGCCTATCATATAGTAAAACCCAGCCCATGG +CCCCTAACAGGGGCCCTCTCAGCCCTCCTAATGACCTCCGGCCTAGCCATGTGATTTCACTTCCACTCCACAACCCTCCT +CATACTAGGCCTACTAACCAACACACTAACCATATACCAATGATGGCGCGATGTAACACGAGAAAGCACATACCAAGGCC +ACCACACACCACCTGTCCAGAAAGGCCTTCGATACGGGATAATCCTATTTATTACCTCAGAAGTTTTTTTCTTCGCAGGA +TTTTTCTGAGCCTTTTACCACTCCAGCCTAGCTCCCACCCCCCAACTAGGGGGACACTGGCCCCCAACAGGCATCACCCC +GCTAAATCCCCTAGAAGTCCCACTCCTAAACACATCCGTATTACTCGCATCAGGGGTATCAATCACCTGAGCTCACCATA +GTCTAATAGAAAACAACCGAAACCAAATAATTCAAGCACTGCTTATTACAATTTTACTGGGTCTCTATTTTACCCTCCTA +CAAGCCTCAGAGTACTTCGAGGTTAAAATATTAGATATTTCCCCTGATACAGGGCTCAATCTTTTTCTTTTTAAAGCAAT +ATTTCTCAAAGTACTTTTCACAGAACTTAAGTTTCATTAAGCACTTCACTAAAAGAAAAGTCTGTGATCTAATAAATTTG +GAAAATATTGAGAATTAGAGCCCCCTCTTAGATATGTACTG +>chr1:5850087-5850571 +GTTGACCTGAACTAATCCCTGCTTGGCGTGTCTGAATAAAGCAGCAACATGCTCAGAGAGGAGGGTGTGCAGTGGCGTCC +CCAGTGGGTCACGCTGCCGTCGCCAACAGTGGATAACGAGGCCGGGGTGGGGGCGCAGCGCTGTCGACTCCGTGGCTCTC +TCAGATCATCCACTGGTTAAAAGTAAAAGACAGCTGAACCCTCATGGGGCCATTCATACAAGTCCCTATTTAAGGAACAA +GTGATTATGCTACCTTTGCACAGTCAGGGTACCGTGGACGTTAAACACATGTCACTGGGCAGGCAGGGCCTCTAATACTG +ACAGTGCTAGAGGTGATGTTTTCGGTAAACAGGCGGGGTAAAAATTTGCCAAGTTCCTTTTTCCATGTCTGTGGATGGCC +TCGCCTCAGGAGATGAGCACCCAGGAAAGAAGTGAGCTCTGTCCCCAGCCCTGCCACAGAAGCCTGGGCTGCACAGGTCC +CACC +>chr1:8909610-8910014 +ACCTTGTGATCCACCCACCTCGGCCTCCCAAACTGCTGGGATTACAGGCGTGAACCACAGCACCTGGCCCCAATACACTC +TTGCATTCCTACGTCCATCTGAGTGCTCACTTCCTGGAGGACCCCACTGACTTAATAATCACATCACAAGACGTCTTACA +CTCATGAACTGTCCCTACATTAGGCCTAAAAACAGATGCAATCCCCGGACGCCTAAACCAAACCACATTCACCGCCACAC +GACCAGGAGTATACTACGGTCAATGGTCAGAAATCTGCAGAGCTAGCCACAGTTTTATACAACCATCTGGAGAGGAGAGG +TGGGCGGGGATTGGATCCTATAGGACCTTGTGGGCCATGGTAAGGGGTTTGAATTCTGTTCTGGTTCTAATGGGAAACCA +TTGC From 7c518b10af705f6d6df7c89e8a8dc4f3c012469c Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 15:12:34 -0400 Subject: [PATCH 085/100] fix: specify tag for cvbio container --- modules/local/prepare_genome.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index faf98df0..9c5914aa 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -94,7 +94,7 @@ process RENAME_DELIM_CONTIGS { """ tag { delim } - container "nciccbr/common_cvbio_3.0.0" + container "nciccbr/common_cvbio_3.0.0:v1" input: path(delim) From 05864035186cf4ea942bce3e6507c4a38e03e0b4 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Sat, 7 Oct 2023 15:17:40 -0400 Subject: [PATCH 086/100] fix: docker container name --- modules/local/prepare_genome.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index 9c5914aa..978a28ec 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -94,7 +94,7 @@ process RENAME_DELIM_CONTIGS { """ tag { delim } - container "nciccbr/common_cvbio_3.0.0:v1" + container "nciccbr/ccbr_cvbio_3.0.0:v1" input: path(delim) From 91535915504e4b49df490815d43cba4ec1d314ed Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 9 Oct 2023 10:16:33 -0400 Subject: [PATCH 087/100] fix: must escape backslashes for nextflow --- modules/local/prepare_genome.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index 978a28ec..399d679a 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -74,7 +74,7 @@ process RENAME_FASTA_CONTIGS { if line.startswith(">"): old_contig = line.strip(">").strip() contig = contig_map[contig] if old_contig in contig_map.keys() else old_contig - outfile.write(f">{config}\n") + outfile.write(f">{config}\\n") else: outfile.write(line) From d444780982a7e1fecc5ecff16511e48b41d9515c Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 9 Oct 2023 11:19:42 -0400 Subject: [PATCH 088/100] ci: only launch full run if requested from manual dispatch --- .github/workflows/build.yml | 10 ++++++++++ 1 file changed, 10 insertions(+) diff --git a/.github/workflows/build.yml b/.github/workflows/build.yml index 207c4633..00a73b34 100644 --- a/.github/workflows/build.yml +++ b/.github/workflows/build.yml @@ -9,6 +9,15 @@ on: branches: - main - develop + workflow_dispatch: + inputs: + test_run: + type: boolean + default: false + required: true + +env: + test_run: ${{ github.event_name == 'workflow_dispatch' && github.event.inputs.test_run }} jobs: build: @@ -33,6 +42,7 @@ jobs: run: | champagne run -profile ci_stub -stub - name: Test run + if: ${{ env.test_run == 'true' }} run: | champagne run -profile ci_test,docker - name: "Upload Artifact" From e006ba83e25f9273c1b9d7dd52d842ab39fcfbd8 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Mon, 9 Oct 2023 11:21:54 -0400 Subject: [PATCH 089/100] ci: always upload artifact --- .github/workflows/build.yml | 1 + 1 file changed, 1 insertion(+) diff --git a/.github/workflows/build.yml b/.github/workflows/build.yml index 00a73b34..84fe1224 100644 --- a/.github/workflows/build.yml +++ b/.github/workflows/build.yml @@ -47,6 +47,7 @@ jobs: champagne run -profile ci_test,docker - name: "Upload Artifact" uses: actions/upload-artifact@v3 + if: always() # run even if previous steps fail with: name: nextflow-log path: .nextflow.log From 7fbb28e09cdbd64d8a164f21330566ec4bad7d8e Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 10 Oct 2023 13:03:33 -0400 Subject: [PATCH 090/100] fix: don't emit metadata, add it in the subwf --- modules/local/prepare_genome.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index 399d679a..e91e6920 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -58,7 +58,7 @@ process RENAME_FASTA_CONTIGS { path(map) output: - tuple val(fasta.baseName), path("*_renamed*"), emit: fasta + path("*_renamed*"), emit: fasta script: def renamed_fasta= "${fasta.getSimpleName()}_renamed.${fasta.getExtension()}" @@ -101,7 +101,7 @@ process RENAME_DELIM_CONTIGS { path(map) output: - tuple val(delim.baseName), path("*_renamed*"), emit: delim + path("*_renamed*"), emit: delim script: def renamed_delim = "${delim.getSimpleName()}_renamed.${delim.getExtension()}" From 7133d34b48919977747f463f9a7b583ede2f6e78 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 10 Oct 2023 14:46:57 -0400 Subject: [PATCH 091/100] fix: switch to ccbr/nf-modules khmer with stub --- modules.json | 5 +++++ .../{nf-core => CCBR}/khmer/uniquekmers/main.nf | 15 +++++++++++---- .../{nf-core => CCBR}/khmer/uniquekmers/meta.yml | 4 ++-- subworkflows/local/prepare_genome.nf | 2 +- 4 files changed, 19 insertions(+), 7 deletions(-) rename modules/{nf-core => CCBR}/khmer/uniquekmers/main.nf (71%) rename modules/{nf-core => CCBR}/khmer/uniquekmers/meta.yml (90%) diff --git a/modules.json b/modules.json index 65ab6d7b..9295813d 100644 --- a/modules.json +++ b/modules.json @@ -19,6 +19,11 @@ "branch": "main", "git_sha": "7a78633cb964d61a663edf2fbff333790658bb00", "installed_by": ["modules"] + }, + "khmer/uniquekmers": { + "branch": "khmer", + "git_sha": "b5b4ba5c89b8a6d5fe117183d198616352fa465a", + "installed_by": ["modules"] } } } diff --git a/modules/nf-core/khmer/uniquekmers/main.nf b/modules/CCBR/khmer/uniquekmers/main.nf similarity index 71% rename from modules/nf-core/khmer/uniquekmers/main.nf rename to modules/CCBR/khmer/uniquekmers/main.nf index 9576034f..4413625d 100644 --- a/modules/nf-core/khmer/uniquekmers/main.nf +++ b/modules/CCBR/khmer/uniquekmers/main.nf @@ -2,10 +2,7 @@ process KHMER_UNIQUEKMERS { tag "$fasta" label 'process_low' - conda "bioconda::khmer=3.0.0a3" - container "${ workflow.containerEngine == 'singularity' && !task.ext.singularity_pull_docker_container ? - 'https://depot.galaxyproject.org/singularity/khmer:3.0.0a3--py37haa7609a_2' : - 'biocontainers/khmer:3.0.0a3--py37haa7609a_2' }" + container 'nciccbr/ccbr_khmer_3.0.0:v1' input: path fasta @@ -35,4 +32,14 @@ process KHMER_UNIQUEKMERS { khmer: \$( unique-kmers.py --version 2>&1 | grep ^khmer | sed 's/^khmer //;s/ .*\$//' ) END_VERSIONS """ + + stub: + """ + touch report.txt kmers.txt + + cat <<-END_VERSIONS > versions.yml + "${task.process}": + khmer: \$( unique-kmers.py --version 2>&1 | grep ^khmer | sed 's/^khmer //;s/ .*\$//' ) + END_VERSIONS + """ } diff --git a/modules/nf-core/khmer/uniquekmers/meta.yml b/modules/CCBR/khmer/uniquekmers/meta.yml similarity index 90% rename from modules/nf-core/khmer/uniquekmers/meta.yml rename to modules/CCBR/khmer/uniquekmers/meta.yml index f9f63972..5ffb451a 100644 --- a/modules/nf-core/khmer/uniquekmers/meta.yml +++ b/modules/CCBR/khmer/uniquekmers/meta.yml @@ -1,5 +1,5 @@ name: "khmer_uniquekmers" -description: In-memory nucleotide sequence k-mer counting, filtering, graph traversal and more +description: In-memory nucleotide sequence k-mer counting, filtering, graph traversal and more. Adapted from nf-core/modules khmer/uniquekhmers. keywords: - khmer - k-mer @@ -39,4 +39,4 @@ output: pattern: "versions.yml" authors: - - "@JoseEspinosa" + - "@kelly-sovacool" diff --git a/subworkflows/local/prepare_genome.nf b/subworkflows/local/prepare_genome.nf index b1787d73..01653333 100644 --- a/subworkflows/local/prepare_genome.nf +++ b/subworkflows/local/prepare_genome.nf @@ -1,6 +1,6 @@ include { BWA_INDEX as BWA_INDEX_BL BWA_INDEX as BWA_INDEX_REF } from "../../modules/CCBR/bwa/index" -include { KHMER_UNIQUEKMERS } from '../../modules/nf-core/khmer/uniquekmers' +include { KHMER_UNIQUEKMERS } from '../../modules/CCBR/khmer/uniquekmers' include { BEDTOOLS_GETFASTA } from '../../modules/nf-core/bedtools/getfasta/main' include { SPLIT_REF_CHROMS RENAME_FASTA_CONTIGS as RENAME_FASTA_CONTIGS_REF From fb3377251472a57277187894b3ef1069d4afc23f Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 10 Oct 2023 15:02:38 -0400 Subject: [PATCH 092/100] feat: download chrom mapping for R64 from https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt --- assets/R64-1-1_ensembl2UCSC.txt | 17 +++++++++++++++++ 1 file changed, 17 insertions(+) create mode 100644 assets/R64-1-1_ensembl2UCSC.txt diff --git a/assets/R64-1-1_ensembl2UCSC.txt b/assets/R64-1-1_ensembl2UCSC.txt new file mode 100644 index 00000000..0a5ee88a --- /dev/null +++ b/assets/R64-1-1_ensembl2UCSC.txt @@ -0,0 +1,17 @@ +I chrI +II chrII +III chrIII +IV chrIV +IX chrIX +Mito chrM +V chrV +VI chrVI +VII chrVII +VIII chrVIII +X chrX +XI chrXI +XII chrXII +XIII chrXIII +XIV chrXIV +XV chrXV +XVI chrXVI From 15398252699aad9992746b47b7fb9e785550e4c7 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 10 Oct 2023 15:02:50 -0400 Subject: [PATCH 093/100] fix: typos --- modules/local/prepare_genome.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index e91e6920..ef0022da 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -73,8 +73,8 @@ process RENAME_FASTA_CONTIGS { for line in infile: if line.startswith(">"): old_contig = line.strip(">").strip() - contig = contig_map[contig] if old_contig in contig_map.keys() else old_contig - outfile.write(f">{config}\\n") + contig = contig_map[old_contig] if old_contig in contig_map.keys() else old_contig + outfile.write(f">{contig}\\n") else: outfile.write(line) From 092b3e02a9fefbd115ef60bb1e016d05223d05c4 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 10 Oct 2023 15:15:07 -0400 Subject: [PATCH 094/100] fix: mitochondrial contig is labeled 'MT' in this genome --- assets/R64-1-1_ensembl2UCSC.txt | 2 +- conf/ci_test.config | 11 ++++++----- conf/test.config | 3 ++- 3 files changed, 9 insertions(+), 7 deletions(-) diff --git a/assets/R64-1-1_ensembl2UCSC.txt b/assets/R64-1-1_ensembl2UCSC.txt index 0a5ee88a..3b5a577c 100644 --- a/assets/R64-1-1_ensembl2UCSC.txt +++ b/assets/R64-1-1_ensembl2UCSC.txt @@ -3,7 +3,7 @@ II chrII III chrIII IV chrIV IX chrIX -Mito chrM +MT chrM V chrV VI chrVI VII chrVII diff --git a/conf/ci_test.config b/conf/ci_test.config index 835b2ea8..83367ea6 100644 --- a/conf/ci_test.config +++ b/conf/ci_test.config @@ -12,7 +12,7 @@ params { genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' blacklist = 'tests/data/test.blacklist' - rename_contigs = 'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' + rename_contigs = 'assets/R64-1-1_ensembl2UCSC.txt' max_cpus = 2 // for GitHub Actions https://docs.github.com/en/actions/using-github-hosted-runners/about-github-hosted-runners#supported-runners-and-hardware-resources @@ -24,11 +24,12 @@ params { // CCBR shared resource paths index_dir = "tests/data" fastq_screen = null - sicer { - species = "sacCer1" // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py - } + sicer.species = "sacCer1" // supported species https://github.com/zanglab/SICER2/blob/master/sicer/lib/GenomeData.py + + deeptools.bin_size = 10000 // this value is only to make bamCoverage run faster. use smaller value for real data. + deeptools.excluded_chroms = 'chrM' run.sicer = false // TODO set to true after https://github.com/CCBR/CHAMPAGNE/issues/109 - run.gem = true // TODO set to true after fixing rename_contigs + run.gem = true } process { diff --git a/conf/test.config b/conf/test.config index 054efe04..89dd0039 100644 --- a/conf/test.config +++ b/conf/test.config @@ -12,12 +12,13 @@ params { genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' - //rename_contigs = 'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' + rename_contigs = 'assets/R64-1-1_ensembl2UCSC.txt' max_cpus = 32 // for interactive node on biowulf max_memory = '120 GB' // for interactive node on biowulf deeptools.bin_size = 10000 // this value is only to make bamCoverage run faster. use smaller value for real data. + deeptools.excluded_chroms = 'chrM' run { qc = false normalize_input = false From 035b74fe1f47c4849e987e874b6339580f516c7c Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Tue, 10 Oct 2023 15:15:55 -0400 Subject: [PATCH 095/100] fix: update cvbio container w/ older java --- modules/local/prepare_genome.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/prepare_genome.nf b/modules/local/prepare_genome.nf index ef0022da..bcd05092 100644 --- a/modules/local/prepare_genome.nf +++ b/modules/local/prepare_genome.nf @@ -94,7 +94,7 @@ process RENAME_DELIM_CONTIGS { """ tag { delim } - container "nciccbr/ccbr_cvbio_3.0.0:v1" + container "nciccbr/ccbr_cvbio_3.0.0:v1.0.1" input: path(delim) From bd991770c5b5b3b36c88567cc04d284c367161e3 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 11 Oct 2023 09:16:54 -0400 Subject: [PATCH 096/100] test: tweak resources --- conf/ci_test.config | 1 - conf/slurmint.config | 4 ++-- conf/test.config | 3 --- conf/test_paired.config | 7 ++----- 4 files changed, 4 insertions(+), 11 deletions(-) diff --git a/conf/ci_test.config b/conf/ci_test.config index 83367ea6..70755d8c 100644 --- a/conf/ci_test.config +++ b/conf/ci_test.config @@ -29,7 +29,6 @@ params { deeptools.bin_size = 10000 // this value is only to make bamCoverage run faster. use smaller value for real data. deeptools.excluded_chroms = 'chrM' run.sicer = false // TODO set to true after https://github.com/CCBR/CHAMPAGNE/issues/109 - run.gem = true } process { diff --git a/conf/slurmint.config b/conf/slurmint.config index 85c68bd9..844b1418 100644 --- a/conf/slurmint.config +++ b/conf/slurmint.config @@ -1,8 +1,8 @@ params { config_profile_name = 'Slurm interactive node on biowulf' - max_memory = '224 GB' + max_memory = '220 GB' max_cpus = 56 - max_time = '72 h' + max_time = '12 h' // CCBR shared resource paths fastq_screen { diff --git a/conf/test.config b/conf/test.config index 89dd0039..35b30cef 100644 --- a/conf/test.config +++ b/conf/test.config @@ -14,9 +14,6 @@ params { blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' rename_contigs = 'assets/R64-1-1_ensembl2UCSC.txt' - max_cpus = 32 // for interactive node on biowulf - max_memory = '120 GB' // for interactive node on biowulf - deeptools.bin_size = 10000 // this value is only to make bamCoverage run faster. use smaller value for real data. deeptools.excluded_chroms = 'chrM' run { diff --git a/conf/test_paired.config b/conf/test_paired.config index 8a5219e3..774aaeb4 100644 --- a/conf/test_paired.config +++ b/conf/test_paired.config @@ -12,13 +12,10 @@ params { genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' - rename_contigs = 'https://raw.githubusercontent.com/dpryan79/ChromosomeMappings/4ead5cc20b6a5d394191cc0bac897759deb02a80/R64-1-1_ensembl2UCSC.txt' - - - max_cpus = 32 // for interactive node on biowulf - max_memory = '120 GB' // for interactive node on biowulf + rename_contigs = 'assets/R64-1-1_ensembl2UCSC.txt' deeptools.bin_size = 10000 // this value is only to make bamCoverage run faster. use smaller value for real data. + deeptools.excluded_chroms = 'chrM' run { qc = false normalize_input = false From 631f854db60990b42a5357210f56c081921e4528 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 11 Oct 2023 09:17:10 -0400 Subject: [PATCH 097/100] fix: make DIY filter for negative GEM peaks --- modules/local/peaks.nf | 52 ++++++++++++++++--------------------- subworkflows/local/peaks.nf | 5 +--- 2 files changed, 23 insertions(+), 34 deletions(-) diff --git a/modules/local/peaks.nf b/modules/local/peaks.nf index 5e17d5b5..a0ad8540 100644 --- a/modules/local/peaks.nf +++ b/modules/local/peaks.nf @@ -255,48 +255,40 @@ process GEM { """ } -process MAKE_CHROM_FILTER { - // to filter out negative peaks from GEM - container "${params.containers.base}" - - input: - path(chrom_sizes) - - output: - path("${chrom_sizes}.filter.txt"), emit: txt - - script: - """ - #!/usr/bin/env python - with open("${chrom_sizes}", "r") as infile: - with open("${chrom_sizes}.filter.txt", "w") as outfile: - for line in infile: - chrom_name, chrom_len = line.strip().split() - outfile.write(f"{chrom_name}\\t1\\t1\\n") - outfile.write(f"{chrom_name}\\t{chrom_len}\\t{chrom_len}\\n") - """ - - stub: - """ - touch ${chrom_sizes}.filter.txt - """ - -} - process FILTER_GEM { tag { meta.id } container "${params.containers.base}" input: - tuple val(meta), path(peak), val(tool), path(chrom_filter) + tuple val(meta), path(peak), val(tool), path(chrom_sizes) output: tuple val(meta), path("${peak}.filtered"), val(tool), emit: peak script: """ - bedtools intersect -v -a ${peak} -b ${chrom_filter} > ${peak}.filtered + #!/usr/bin/env python + + chrom_ends = dict() + with open('${chrom_sizes}', 'r') as chrom_file: + for line in chrom_file: + chrom, start, end = line.split() + chrom_ends[chrom] = int(end) + + count_bad_peaks = 0 + with open('${peak}', 'r') as infile: + with open('${peak}.filtered', 'w') as outfile: + for line in infile: + line_split = line.split() + chrom = line_split[0] + start = int(line_split[1]) + end = int(line_split[2]) + if start > 0 and end < chrom_ends[chrom]: + outfile.write(line) + else: + count_bad_peaks += 1 + print(f"Filtered out {count_bad_peaks} peaks") """ stub: """ diff --git a/subworkflows/local/peaks.nf b/subworkflows/local/peaks.nf index db33244b..dee68a6a 100644 --- a/subworkflows/local/peaks.nf +++ b/subworkflows/local/peaks.nf @@ -5,7 +5,6 @@ include { CALC_GENOME_FRAC SICER CONVERT_SICER GEM - MAKE_CHROM_FILTER FILTER_GEM FRACTION_IN_PEAKS CONCAT_FRIPS @@ -96,9 +95,7 @@ workflow CALL_PEAKS { if (params.run.gem) { ch_gem | GEM GEM.out.peak - .combine( - MAKE_CHROM_FILTER(chrom_sizes).txt - ) | FILTER_GEM + .combine(chrom_sizes) | FILTER_GEM ch_peaks = ch_peaks.mix(FILTER_GEM.out.peak) } From 44b373536a02b139de88f03a44c2669931c5303c Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 11 Oct 2023 09:53:34 -0400 Subject: [PATCH 098/100] fix: parse chrom lengths --- modules/local/peaks.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/modules/local/peaks.nf b/modules/local/peaks.nf index a0ad8540..0aadb9f8 100644 --- a/modules/local/peaks.nf +++ b/modules/local/peaks.nf @@ -273,7 +273,7 @@ process FILTER_GEM { chrom_ends = dict() with open('${chrom_sizes}', 'r') as chrom_file: for line in chrom_file: - chrom, start, end = line.split() + chrom, end = line.split() chrom_ends[chrom] = int(end) count_bad_peaks = 0 From 757e06805f95f4283921e41e5b7d59dfa8d41100 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 11 Oct 2023 10:31:26 -0400 Subject: [PATCH 099/100] test: consolidate test & test_paired into one profile --- assets/samplesheet_test.csv | 6 ++--- assets/samplesheet_test_paired.csv | 7 ----- conf/ci_stub.config | 2 +- conf/ci_test.config | 2 +- conf/test.config | 9 ++++--- conf/test_paired.config | 42 ------------------------------ nextflow.config | 3 --- 7 files changed, 10 insertions(+), 61 deletions(-) delete mode 100644 assets/samplesheet_test_paired.csv delete mode 100644 conf/test_paired.config diff --git a/assets/samplesheet_test.csv b/assets/samplesheet_test.csv index 4f0585ca..58a50b71 100644 --- a/assets/samplesheet_test.csv +++ b/assets/samplesheet_test.csv @@ -1,7 +1,7 @@ sample,fastq_1,fastq_2,antibody,control -SPT5_T0_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822153_1.fastq.gz,,SPT5,SPT5_INPUT_REP1 +SPT5_T0_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822153_1.fastq.gz,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822153_2.fastq.gz,SPT5,SPT5_INPUT_REP1 SPT5_T0_REP2,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822154_1.fastq.gz,,SPT5,SPT5_INPUT_REP2 -SPT5_T15_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822157_1.fastq.gz,,SPT5,SPT5_INPUT_REP1 +SPT5_T15_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822157_1.fastq.gz,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822157_2.fastq.gz,SPT5,SPT5_INPUT_REP1 SPT5_T15_REP2,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822158_1.fastq.gz,,SPT5,SPT5_INPUT_REP2 -SPT5_INPUT_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/testdata/SRR5204809_Spt5-ChIP_Input1_SacCer_ChIP-Seq_ss100k_R1.fastq.gz,,, +SPT5_INPUT_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/testdata/SRR5204809_Spt5-ChIP_Input1_SacCer_ChIP-Seq_ss100k_R1.fastq.gz,https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/testdata/SRR5204809_Spt5-ChIP_Input1_SacCer_ChIP-Seq_ss100k_R2.fastq.gz,, SPT5_INPUT_REP2,https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/testdata/SRR5204810_Spt5-ChIP_Input2_SacCer_ChIP-Seq_ss100k_R1.fastq.gz,,, diff --git a/assets/samplesheet_test_paired.csv b/assets/samplesheet_test_paired.csv deleted file mode 100644 index 58a50b71..00000000 --- a/assets/samplesheet_test_paired.csv +++ /dev/null @@ -1,7 +0,0 @@ -sample,fastq_1,fastq_2,antibody,control -SPT5_T0_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822153_1.fastq.gz,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822153_2.fastq.gz,SPT5,SPT5_INPUT_REP1 -SPT5_T0_REP2,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822154_1.fastq.gz,,SPT5,SPT5_INPUT_REP2 -SPT5_T15_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822157_1.fastq.gz,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822157_2.fastq.gz,SPT5,SPT5_INPUT_REP1 -SPT5_T15_REP2,https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/testdata/SRR1822158_1.fastq.gz,,SPT5,SPT5_INPUT_REP2 -SPT5_INPUT_REP1,https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/testdata/SRR5204809_Spt5-ChIP_Input1_SacCer_ChIP-Seq_ss100k_R1.fastq.gz,https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/testdata/SRR5204809_Spt5-ChIP_Input1_SacCer_ChIP-Seq_ss100k_R2.fastq.gz,, -SPT5_INPUT_REP2,https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/testdata/SRR5204810_Spt5-ChIP_Input2_SacCer_ChIP-Seq_ss100k_R1.fastq.gz,,, diff --git a/conf/ci_stub.config b/conf/ci_stub.config index 795e9db5..80837384 100644 --- a/conf/ci_stub.config +++ b/conf/ci_stub.config @@ -3,7 +3,7 @@ params { config_profile_description = 'Minimal test dataset with blank references to run stubs with continuous integration' outdir = 'results/test' - input = 'assets/samplesheet_test_paired.csv' // adapted from https://github.com/nf-core/test-datasets/blob/chipseq/samplesheet/v2.0/samplesheet_test.csv + input = 'assets/samplesheet_test.csv' // adapted from https://github.com/nf-core/test-datasets/blob/chipseq/samplesheet/v2.0/samplesheet_test.csv genome = 'test' max_cpus = 2 // for GitHub Actions https://docs.github.com/en/actions/using-github-hosted-runners/about-github-hosted-runners#supported-runners-and-hardware-resources diff --git a/conf/ci_test.config b/conf/ci_test.config index 70755d8c..757cf4a0 100644 --- a/conf/ci_test.config +++ b/conf/ci_test.config @@ -3,7 +3,7 @@ params { config_profile_description = 'Minimal test dataset with blank references to run stubs with continuous integration' outdir = 'results/test' - input = 'assets/samplesheet_test_paired.csv' // adapted from https://github.com/nf-core/test-datasets/blob/chipseq/samplesheet/v2.0/samplesheet_test.csv + input = 'assets/samplesheet_test.csv' // adapted from https://github.com/nf-core/test-datasets/blob/chipseq/samplesheet/v2.0/samplesheet_test.csv genome = 'custom_genome' read_length = 50 diff --git a/conf/test.config b/conf/test.config index 35b30cef..207506d2 100644 --- a/conf/test.config +++ b/conf/test.config @@ -1,9 +1,9 @@ params { - config_profile_name = 'Test single-end' - config_profile_description = 'Minimal test dataset to check pipeline function' + config_profile_name = 'Test paired-end and single-end mixed' + config_profile_description = 'Minimal test dataset to check paired & single end handling' outdir = 'results/test' - input = 'assets/samplesheet_test.csv' // adapted from https://github.com/nf-core/test-datasets/blob/chipseq/samplesheet/v2.0/samplesheet_test.csv + input = 'assets/samplesheet_test.csv' // adapted from 'https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/samplesheet/v2.0/samplesheet_test.csv' genome = 'custom_genome' read_length = 50 @@ -21,7 +21,7 @@ params { normalize_input = false call_peaks = true gem = true - sicer = false // TODO set to true after https://github.com/CCBR/CHAMPAGNE/issues/109 + sicer = false macs_broad = true macs_narrow = true } @@ -38,4 +38,5 @@ report { enabled = true overwrite = true file = "${params.outdir}/pipeline_info/execution_report.html" + } diff --git a/conf/test_paired.config b/conf/test_paired.config deleted file mode 100644 index 774aaeb4..00000000 --- a/conf/test_paired.config +++ /dev/null @@ -1,42 +0,0 @@ -params { - config_profile_name = 'Test paired-end and single-end mixed' - config_profile_description = 'Minimal test dataset to check paired & single end handling' - - outdir = 'results/test_paired' - input = 'assets/samplesheet_test_paired.csv' // adapted from 'https://raw.githubusercontent.com/nf-core/test-datasets/chipseq/samplesheet/v2.0/samplesheet_test.csv' - - genome = 'custom_genome' - read_length = 50 - - // Genome references - genome_fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' - genes_gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' - blacklist = '/data/CCBR_Pipeliner/db/PipeDB/Indices/hg38_basic/indexes/hg38.blacklist_v3.chrM.chr_rDNA.fa' - rename_contigs = 'assets/R64-1-1_ensembl2UCSC.txt' - - deeptools.bin_size = 10000 // this value is only to make bamCoverage run faster. use smaller value for real data. - deeptools.excluded_chroms = 'chrM' - run { - qc = false - normalize_input = false - call_peaks = true - gem = true - sicer = false - macs_broad = true - macs_narrow = true - } - sicer { - species = "sacCer1" - } -} -dag { - enabled = true - overwrite = true - file = "assets/dag.png" -} -report { - enabled = true - overwrite = true - file = "${params.outdir}/pipeline_info/execution_report.html" - -} diff --git a/nextflow.config b/nextflow.config index 010c27a7..75d09d69 100644 --- a/nextflow.config +++ b/nextflow.config @@ -103,9 +103,6 @@ profiles { test { includeConfig "conf/test.config" } - test_paired { - includeConfig "conf/test_paired.config" - } test_mm10 { includeConfig "conf/test_mm10.config" } From 4abed35ff4a7e822eb178d6ce546d67d8559f5b6 Mon Sep 17 00:00:00 2001 From: Kelly Sovacool Date: Wed, 11 Oct 2023 10:34:33 -0400 Subject: [PATCH 100/100] docs: update w/ new custom genome option --- CHANGELOG.md | 3 ++- README.md | 23 +++++++++++++++-------- assets/dag.png | Bin 108630 -> 857782 bytes 3 files changed, 17 insertions(+), 9 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 453ba1ea..cccafd5e 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -8,7 +8,8 @@ - Fraction in Peaks (FRiP) (#89) - Jaccard index (#92) - Histogram of peak widths (#92) -- Added an option to use a custom reference from a genome fasta, gtf, and blacklist file. +- Added support for paired-end reads. (#105) +- Added an option to use a custom reference from a genome fasta, gtf, and blacklist file. (#105) ### Bug fixes diff --git a/README.md b/README.md index 1e041894..d2370feb 100644 --- a/README.md +++ b/README.md @@ -15,28 +15,35 @@ You can run champagne from the command line. The CLI includes helper steps for execution on supported high performance computing clusters including Biowulf and FRCE. -Run the test dataset using the test profile: +Run preview to view processes that will run: ```sh -champagne run -profile test,singularity +champagne run -profile ci_stub -preview ``` -or explicitly specify the output directory and input: +Launch a stub run to view processes that will run and download containers: ```sh -champagne run -profile singularity --outdir results/test --input assets/samplesheet_test.csv +champagne run -profile ci_stub,singularity -stub ``` -Run preview to view that steps that will run without actually executing any code: +Run the test dataset using the test profile: ```sh -champagne run -profile ci_stub -preview +champagne run -profile test,singularity +``` + +or explicitly specify the output directory and input: + +```sh +champagne run -profile singularity --outdir results/test --input assets/samplesheet_test.csv ``` -Launch a stub run to 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