Flexible AdapterRemoval

.travis.yml

@@ -40,6 +40,12 @@ script:
   - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker --pairedEnd --saveReference
   # Run the basic pipeline with single end data (pretending its single end actually)
   - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker --singleEnd --bwa_index results/reference_genome/bwa_index/bwa_index/
+  # Run the basic pipeline with paired end data without collapsing
+  - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker --pairedEnd --skip_collapse --saveReference
+  # Run the basic pipeline with paired end data without trimming
+  - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker --pairedEnd --skip_trim --saveReference
+   # Run the basic pipeline with paired end data without adapterRemoval
+  - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker --pairedEnd --skip_adapterremoval --saveReference
   # Run the same pipeline testing optional step: fastp, complexity 
   - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker --pairedEnd --complexity_filter --bwa_index results/reference_genome/bwa_index/bwa_index/
   # Test BAM Trimming

CHANGELOG.md

@@ -10,6 +10,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.
 
 * [#152](https://github.com/nf-core/eager/pull/152) - Clarified `--complexity_filter` flag to be specifically for poly G trimming.
 * [#155](https://github.com/nf-core/eager/pull/155) - Added [Dedup log to output folders](https://github.com/nf-core/eager/issues/154)
+* [#159](https://github.com/nf-core/eager/pull/159) - Added Possibility to skip AdapterRemoval, skip merging, skip trimming fixing [#64](https://github.com/nf-core/eager/issues/64),[#137](https://github.com/nf-core/eager/issues/137) - thanks to @maxibor, @jfy133
 
 ### `Fixed`
 

docs/usage.md

@@ -283,12 +283,16 @@ This part of the documentation contains a list of user-adjustable parameters in
 
 ## Step skipping parameters
 
-Some of the steps in the pipeline can be executed optionally. If you specify specific steps to be skipped, there won't be any output related to these modules. 
+Some of the steps in the pipeline can be executed optionally. If you specify specific steps to be skipped, there won't be any output related to these modules.
 
 ### `--skip_preseq`
 
 Turns off the computation of library complexity estimation.  
 
+### `--skip_adapterremoval`
+
+Turns off adaptor trimming and paired-end read merging. Equivalent to setting both `--skip_collapse` and `--skip_trim`.
+
 ### `--skip_damage_calculation`
 
 Turns off the DamageProfiler module to compute DNA damage profiles. 
@@ -333,6 +337,24 @@ Defines the minimum read quality per base that is required for a base to be kept
 ### `--clip_min_adap_overlap` 1
 Sets the minimum overlap between two reads when read merging is performed. Default is set to `1` base overlap.
 
+### `--skip_collapse`
+
+Turns off the paired-end read merging.
+
+For example
+```bash
+--pairedEnd --skip_collapse  --reads '*.fastq'
+```
+
+### `--skip_trim`
+
+Turns off adaptor and quality trimming.
+
+For example:
+```bash
+--pairedEnd --skip_trim  --reads '*.fastq'
+```
+
 ## Read Mapping Parameters
 
 ## BWA (default)

main.nf

@@ -44,6 +44,7 @@ def helpMessage() {
       --saveReference               Saves reference genome indices for later reusage
 
     Skipping                        Skip any of the mentioned steps
+      --skip_adapterremoval         
       --skip_preseq
       --skip_damage_calculation
       --skip_qualimap
@@ -59,6 +60,8 @@ def helpMessage() {
       --clip_readlength             Specify read minimum length to be kept for downstream analysis
       --clip_min_read_quality       Specify minimum base quality for not trimming off bases
       --min_adap_overlap            Specify minimum adapter overlap
+      --skip_collapse               Skip merging forward and reverse reads together. (Only for PE samples)
+      --skip_trim                   Skip adaptor and quality trimming
 
     BWA Mapping
       --bwaalnn                     Specify the -n parameter for BWA aln.
@@ -145,6 +148,7 @@ params.email = false
 params.plaintext_email = false
 
 // Skipping parts of the pipeline for impatient users
+params.skip_adapterremoval = false
 params.skip_preseq = false
 params.skip_damage_calculation = false
 params.skip_qualimap = false
@@ -160,6 +164,8 @@ params.clip_reverse_adaptor = "AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTA"
 params.clip_readlength = 30
 params.clip_min_read_quality = 20
 params.min_adap_overlap = 1
+params.skip_collapse = false
+params.skip_trim = false
 
 //Read mapping parameters (default = BWA aln default)
 params.bwaalnn = 0.04
@@ -258,6 +264,10 @@ if( params.singleEnd || params.pairedEnd || params.bam){
     exit 1, "Please specify either --singleEnd, --pairedEnd to execute the pipeline on FastQ files and --bam for previously processed BAM files!"
 }
 
+//Validate that skip_collapse is only set to True for pairedEnd reads!
+if (params.skip_collapse  && params.singleEnd){
+    exit 1, "--skip_collapse can only be set for pairedEnd samples!"
+}
 
 //AWSBatch sanity checking
 if(workflow.profile == 'awsbatch'){
@@ -343,6 +353,8 @@ summary['Fasta Ref']    = params.fasta
 summary['BAM Index Type'] = (params.large_ref == "") ? 'BAI' : 'CSI'
 if(params.bwa_index) summary['BWA Index'] = params.bwa_index
 summary['Data Type']    = params.singleEnd ? 'Single-End' : 'Paired-End'
+summary['Skip Collapsing'] = params.skip_collapse ? 'Yes' : 'No'
+summary['Skip Trimming']  = params.skip_trim  ? 'Yes' : 'No' 
 summary['Max Memory']   = params.max_memory
 summary['Max CPUs']     = params.max_cpus
 summary['Max Time']     = params.max_time
@@ -362,7 +374,7 @@ if(workflow.profile == 'awsbatch'){
    summary['AWS Queue'] = params.awsqueue
 }
 if(params.email) summary['E-mail Address'] = params.email
-log.info summary.collect { k,v -> "${k.padRight(15)}: $v" }.join("\n")
+log.info summary.collect { k,v -> "${k.padRight(35)}: $v" }.join("\n")
 log.info "========================================="
 
 
@@ -501,8 +513,7 @@ process convertBam {
     file bam from ch_bam_to_fastq_convert
 
     output:
-    set val("${base}"), file("*.fastq.gz") into (ch_read_files_converted_fastqc, ch_read_files_converted_fastp)
-    file("*.fastq.gz") into (ch_read_files_converted_mapping_bwa, ch_read_files_converted_mapping_cm, ch_read_files_converted_mapping_bwamem)
+    set val("${base}"), file("*.fastq.gz") into (ch_read_files_converted_fastqc, ch_read_files_converted_fastp, ch_read_files_converted_mapping_bwa, ch_read_files_converted_mapping_cm, ch_read_files_converted_mapping_bwamem)
 
     script:
     base = "${bam.baseName}"
@@ -568,60 +579,87 @@ process fastp {
 /*
  * STEP 2 - Adapter Clipping / Read Merging
  */
-
-
+//Initialize empty channel if we skip adapterremoval entirely
+if(params.skip_adapterremoval) {
+    //No logs if no AR is run
+    ch_adapterremoval_logs = Channel.empty()
+    //Either coming from complexity filtering, or directly use reads normally directed to clipping first and push them through to the other channels downstream! 
+    ch_clipped_reads_complexity_filtered_poly_g.mix(ch_read_files_clip).into { ch_clipped_reads;ch_clipped_reads_for_fastqc;ch_clipped_reads_circularmapper;ch_clipped_reads_bwamem }
+} else {
 process adapter_removal {
     tag "$name"
     publishDir "${params.outdir}/read_merging", mode: 'copy'
 
-    when: !params.bam
+    when: !params.bam && !params.skip_adapterremoval
 
     input:
     set val(name), file(reads) from ( params.complexity_filter_poly_g ? ch_clipped_reads_complexity_filtered_poly_g : ch_read_files_clip )
 
     output:
-    file "*.combined*.gz" into (ch_clipped_reads, ch_clipped_reads_for_fastqc,ch_clipped_reads_circularmapper,ch_clipped_reads_bwamem)
-    file "*.settings" into ch_adapterremoval_logs
+    set val(base), file("output/*.gz") into (ch_clipped_reads,ch_clipped_reads_for_fastqc,ch_clipped_reads_circularmapper,ch_clipped_reads_bwamem)
+    file("*.settings") into ch_adapterremoval_logs
 
     script:
-    prefix = reads[0].toString() - ~/(_R1)?(_trimmed)?(_val_1)?(\.fq)?(\.fastq)?(\.gz)?$/
-    //Readprefixing only required for PE data with merging
-    fixprefix = (params.singleEnd) ? "" : "AdapterRemovalFixPrefix ${prefix}.combined.fq.gz ${prefix}.combined.prefixed.fq.gz"
+    base = reads[0].baseName
+    //This checks whether we skip trimming and defines a variable respectively
+    trim_me = params.skip_trim ? '' : "--trimns --trimqualities --adapter1 ${params.clip_forward_adaptor} --adapter2 ${params.clip_reverse_adaptor} --minlength ${params.clip_readlength} --minquality ${params.clip_min_read_quality} --minadapteroverlap ${params.min_adap_overlap}"
+    collapse_me = params.skip_collapse ? '' : '--collapse'
 
-    if( !params.singleEnd ){
+    //PE mode, dependent on trim_me and collapse_me the respective procedure is run or not :-) 
+    if (!params.singleEnd && !params.skip_collapse && !params.skip_trim){
     """
-    AdapterRemoval --file1 ${reads[0]} --file2 ${reads[1]} --basename ${prefix} --gzip --threads ${task.cpus} --trimns --trimqualities --adapter1 ${params.clip_forward_adaptor} --adapter2 ${params.clip_reverse_adaptor} --minlength ${params.clip_readlength} --minquality ${params.clip_min_read_quality} --minadapteroverlap ${params.min_adap_overlap} --collapse
+    mkdir -p output
+    AdapterRemoval --file1 ${reads[0]} --file2 ${reads[1]} --basename ${base} ${trim_me} --gzip --threads ${task.cpus} ${collapse_me}
     #Combine files
-    zcat *.collapsed.gz *.collapsed.truncated.gz *.singleton.truncated.gz *.pair1.truncated.gz *.pair2.truncated.gz | gzip > ${prefix}.combined.fq.gz
-    ${fixprefix}
-    rm ${prefix}.combined.fq.gz
+    zcat *.collapsed.gz *.collapsed.truncated.gz *.singleton.truncated.gz *.pair1.truncated.gz *.pair2.truncated.gz | gzip > output/${base}.combined.fq.gz
+    """
+    //PE, don't collapse, but trim reads
+    } else if (!params.singleEnd && params.skip_collapse && !params.skip_trim) {
+    """
+    mkdir -p output
+    AdapterRemoval --file1 ${reads[0]} --file2 ${reads[1]} --basename ${base} --gzip --threads ${task.cpus} ${trim_me} ${collapse_me}
+    mv ${base}.pair*.truncated.gz output/
+    """
+    //PE, collapse, but don't trim reads
+    } else if (!params.singleEnd && !params.skip_collapse && params.skip_trim) {
+    """
+    mkdir -p output
+    AdapterRemoval --file1 ${reads[0]} --file2 ${reads[1]} --basename ${base} --gzip --threads ${task.cpus} --basename ${base} ${collapse_me} ${trim_me}
+
+    mv ${base}.pair*.truncated.gz output/
     """
     } else {
+    //SE, collapse not possible, trim reads
     """
-    AdapterRemoval --file1 ${reads[0]} --basename ${prefix} --gzip --threads ${task.cpus} --trimns --trimqualities --adapter1 ${params.clip_forward_adaptor} --minlength ${params.clip_readlength} --minquality ${params.clip_min_read_quality} 
-    # Pseudo-Combine
-    mv *.truncated.gz ${prefix}.combined.fq.gz
+    mkdir -p output
+    AdapterRemoval --file1 ${reads[0]} --basename ${base} --gzip --threads ${task.cpus} ${trim_me}
+
+    mv *.truncated.gz output/
     """
     }
 }
+}
+
+
 
 /*
- * STEP 2.1 - FastQC after clipping/merging (if applied!)
- */
+* STEP 2.1 - FastQC after clipping/merging (if applied!)
+*/
 process fastqc_after_clipping {
-    tag "${reads[0].baseName}"
+    tag "${prefix}"
     publishDir "${params.outdir}/FastQC/after_clipping", mode: 'copy',
         saveAs: {filename -> filename.indexOf(".zip") > 0 ? "zips/$filename" : "$filename"}
 
-    when: !params.bam
+    when: !params.bam && !params.skip_adapterremoval
 
     input:
-    file(reads) from ch_clipped_reads_for_fastqc
+    set val(name), file(reads) from ch_clipped_reads_for_fastqc
 
     output:
     file "*_fastqc.{zip,html}" optional true into ch_fastqc_after_clipping
 
     script:
+    prefix = reads[0].toString().tokenize('.')[0]
     """
     fastqc -q $reads
     """
@@ -638,7 +676,8 @@ process bwa {
     when: !params.circularmapper && !params.bwamem
 
     input:
-    file(reads) from ch_clipped_reads.mix(ch_read_files_converted_mapping_bwa)
+    set val(name), file(reads) from ch_clipped_reads.mix(ch_read_files_converted_mapping_bwa)
+
     file index from ch_bwa_index.first()
 
 
@@ -648,14 +687,28 @@ process bwa {
 
 
     script:
-    prefix = reads[0].toString() - ~/(_R1)?(\.combined\.)?(prefixed)?(_trimmed)?(_val_1)?(\.fq)?(\.fastq)?(\.gz)?$/
-    fasta = "${index}/*.fasta" 
+    fasta = "${index}/*.fasta"
     size = "${params.large_ref}" ? '-c' : ''
+
+    //PE data without merging, PE data without any AR applied
+    if (!params.singleEnd && (params.skip_collapse || params.skip_adapterremoval)){
+    prefix = reads[0].toString().tokenize('.')[0]
     """ 
-    bwa aln -t ${task.cpus} $fasta $reads -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f "${reads.baseName}.sai"
-    bwa samse -r "@RG\\tID:ILLUMINA-${prefix}\\tSM:${prefix}\\tPL:illumina" $fasta "${reads.baseName}".sai $reads | samtools sort -@ ${task.cpus} -O bam - > "${prefix}".sorted.bam
+    bwa aln -t ${task.cpus} $fasta ${reads[0]} -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f ${prefix}.r1.sai
+    bwa aln -t ${task.cpus} $fasta ${reads[1]} -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f ${prefix}.r2.sai
+    bwa sampe -r "@RG\\tID:ILLUMINA-${prefix}\\tSM:${prefix}\\tPL:illumina" $fasta ${prefix}.r1.sai ${prefix}.r2.sai ${reads[0]} ${reads[1]} | samtools sort -@ ${task.cpus} -O bam - > ${prefix}.sorted.bam
     samtools index "${size}" "${prefix}".sorted.bam
     """
+    } else {
+    //PE collapsed, or SE data 
+    prefix = reads[0].toString().tokenize('.')[0]
+    """ 
+    bwa aln -t ${task.cpus} $fasta $reads -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f ${prefix}.sai
+    bwa samse -r "@RG\\tID:ILLUMINA-${prefix}\\tSM:${prefix}\\tPL:illumina" $fasta ${prefix}.sai $reads | samtools sort -@ ${task.cpus} -O bam - > "${prefix}".sorted.bam
+    samtools index "${size}" "${prefix}".sorted.bam
+    """
+    }
+
 }
 
 process circulargenerator{
@@ -694,7 +747,7 @@ process circularmapper{
     when: params.circularmapper
 
     input:
-    file reads from ch_clipped_reads_circularmapper.mix(ch_read_files_converted_mapping_cm)
+    set val(name), file(reads) from ch_clipped_reads_circularmapper.mix(ch_read_files_converted_mapping_cm)
     file index from ch_circularmapper_indices.first()
 
     output:
@@ -703,17 +756,31 @@ process circularmapper{
 
     script:
     filter = "${params.circularfilter}" ? '' : '-f true -x false'
-    prefix = reads[0].toString() - ~/(_R1)?(\.combined\.)?(prefixed)?(_trimmed)?(_val_1)?(\.fq)?(\.fastq)?(\.gz)?$/
+
     fasta = "${index}/*_*.fasta"
     size = "${params.large_ref}" ? '-c' : ''
 
+    if (!params.singleEnd && params.skip_collapse ){
+    prefix = reads[0].toString().tokenize('.')[0]
     """ 
-    bwa aln -t ${task.cpus} $fasta $reads -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f "${reads.baseName}.sai"
-    bwa samse -r "@RG\\tID:ILLUMINA-${prefix}\\tSM:${prefix}\\tPL:illumina" $fasta "${reads.baseName}".sai $reads > tmp.out
+    bwa aln -t ${task.cpus} $fasta ${reads[0]} -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f ${prefix}.r1.sai
+    bwa aln -t ${task.cpus} $fasta ${reads[1]} -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f ${prefix}.r2.sai
+    bwa sampe -r "@RG\\tID:ILLUMINA-${prefix}\\tSM:${prefix}\\tPL:illumina" $fasta ${prefix}.r1.sai ${prefix}.r2.sai ${reads[0]} ${reads[1]} > tmp.out
+    realignsamfile -e ${params.circularextension} -i tmp.out -r $fasta $filter 
+    samtools sort -@ ${task.cpus} -O bam tmp_realigned.bam > ${prefix}.sorted.bam
+    samtools index "${size}" ${prefix}.sorted.bam
+    """
+    } else {
+    prefix = reads[0].toString().tokenize('.')[0]
+    """ 
+    bwa aln -t ${task.cpus} $fasta $reads -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f ${prefix}.sai
+    bwa samse -r "@RG\\tID:ILLUMINA-${prefix}\\tSM:${prefix}\\tPL:illumina" $fasta ${prefix}.sai $reads > tmp.out
     realignsamfile -e ${params.circularextension} -i tmp.out -r $fasta $filter 
     samtools sort -@ ${task.cpus} -O bam tmp_realigned.bam > "${prefix}".sorted.bam
     samtools index "${size}" "${prefix}".sorted.bam
     """
+    }
+
 }
 
 process bwamem {
@@ -723,7 +790,7 @@ process bwamem {
     when: params.bwamem && !params.circularmapper
 
     input:
-    file(reads) from ch_clipped_reads_bwamem.mix(ch_read_files_converted_mapping_bwamem)
+    set val(name), file(reads) from ch_clipped_reads_bwamem.mix(ch_read_files_converted_mapping_bwamem)
     file index from ch_bwa_index_bwamem.first()
 
     output:
@@ -735,10 +802,19 @@ process bwamem {
     prefix = reads[0].toString() - ~/(_R1)?(\.combined\.)?(prefixed)?(_trimmed)?(_val_1)?(\.fq)?(\.fastq)?(\.gz)?$/
     fasta = "${index}/*.fasta"
     size = "${params.large_ref}" ? '-c' : ''
+
+    if (!params.singleEnd && params.skip_collapse){
+    """
+    bwa mem -t ${task.cpus} $fasta ${reads[0]} ${reads[1]} -R "@RG\\tID:ILLUMINA-${prefix}\\tSM:${prefix}\\tPL:illumina" | samtools sort -@ ${task.cpus} -O bam - > "${prefix}".sorted.bam
+    samtools index "${size}" -@ ${task.cpus} "${prefix}".sorted.bam
+    """
+    } else {
     """
     bwa mem -t ${task.cpus} $fasta $reads -R "@RG\\tID:ILLUMINA-${prefix}\\tSM:${prefix}\\tPL:illumina" | samtools sort -@ ${task.cpus} -O bam - > "${prefix}".sorted.bam
-    samtools index  "${size}" -@ ${task.cpus} "${prefix}".sorted.bam
+    samtools index "${size}" -@ ${task.cpus} "${prefix}".sorted.bam
     """
+    }
+
 }
 
 /*

nextflow.config

@@ -14,6 +14,7 @@ params {
 
   //Pipeline options
   aligner = 'bwa'
+
   saveReference = false
   saveTrimmed = true
   saveAlignedIntermediates = false
@@ -30,6 +31,11 @@ params {
   complexity_filter_poly_g_min = 10
   trim_bam = false
 
+  //Skipping adapterremoval, trimming or collapsing defaults
+  skip_adapterremoval = false
+  skip_trim = false
+  skip_adapterremoval = false
+
   // AWS Batch
   awsqueue = false
   awsregion = 'eu-west-1'
@@ -38,6 +44,7 @@ params {
   custom_config_version = 'master'
 }
 
+
 // Load base.config by default for all pipelines
 includeConfig 'conf/base.config'