From 9b3894f3b7984027b8349f2f4cad326aaddf61cd Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 14 Sep 2021 18:57:26 +0200 Subject: [PATCH 01/25] Post release bump --- .github/workflows/ci.yml | 4 ++-- CHANGELOG.md | 10 ++++++++++ Dockerfile | 4 ++-- environment.yml | 2 +- nextflow.config | 4 ++-- 5 files changed, 17 insertions(+), 7 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 9ad407e8b..21b7ae020 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -37,13 +37,13 @@ jobs: - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t nfcore/eager:2.4.0 + run: docker build --no-cache . -t nfcore/eager:dev - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull nfcore/eager:dev - docker tag nfcore/eager:dev nfcore/eager:2.4.0 + docker tag nfcore/eager:dev nfcore/eager:dev - name: Install Nextflow env: diff --git a/CHANGELOG.md b/CHANGELOG.md index bf8c396b0..18f35ae3a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,6 +3,16 @@ The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). +## [dev] + +### `Added` + +### `Fixed` + +### `Dependencies` + +### `Deprecated` + ## [2.4.0] - 20201-09-14 ### `Added` diff --git a/Dockerfile b/Dockerfile index 20832cddc..7087c402a 100644 --- a/Dockerfile +++ b/Dockerfile @@ -7,7 +7,7 @@ COPY environment.yml / RUN conda env create --quiet -f /environment.yml && conda clean -a # Add conda installation dir to PATH (instead of doing 'conda activate') -ENV PATH /opt/conda/envs/nf-core-eager-2.4.0/bin:$PATH +ENV PATH /opt/conda/envs/nf-core-eager-2.4.1dev/bin:$PATH # Dump the details of the installed packages to a file for posterity -RUN conda env export --name nf-core-eager-2.4.0 > nf-core-eager-2.4.0.yml \ No newline at end of file +RUN conda env export --name nf-core-eager-2.4.1dev > nf-core-eager-2.4.1dev.yml \ No newline at end of file diff --git a/environment.yml b/environment.yml index e8a171f61..3b007e300 100644 --- a/environment.yml +++ b/environment.yml @@ -1,6 +1,6 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml -name: nf-core-eager-2.4.0 +name: nf-core-eager-2.4.1dev channels: - conda-forge - bioconda diff --git a/nextflow.config b/nextflow.config index a1952c731..f86b9dbd8 100644 --- a/nextflow.config +++ b/nextflow.config @@ -278,7 +278,7 @@ params { // Container slug. Stable releases should specify release tag! // Developmental code should specify :dev -process.container = 'nfcore/eager:2.4.0' +process.container = 'nfcore/eager:dev' // Load base.config by default for all pipelines includeConfig 'conf/base.config' @@ -408,7 +408,7 @@ manifest { description = 'A fully reproducible and state-of-the-art ancient DNA analysis pipeline' mainScript = 'main.nf' nextflowVersion = '>=20.07.1' - version = '2.4.0' + version = '2.4.1dev' } // Function to ensure that resource requirements don't go beyond From 19b14e0150f695ec87579ce0479cb34c3d9e17e1 Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Thu, 16 Sep 2021 09:49:24 +0200 Subject: [PATCH 02/25] :facepalm: Add the missing release name of previous release --- CHANGELOG.md | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 18f35ae3a..dbbc3ce01 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -13,7 +13,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Deprecated` -## [2.4.0] - 20201-09-14 +## [2.4.0] - Wangen - 20201-09-14 ### `Added` @@ -178,7 +178,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Deprecated` -## [2.3.0] - 2021-01-11 - "Aalen" +## [2.3.0] - Aalen - 2021-01-11 ### `Added` @@ -311,7 +311,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. - Latest version of Sex.DetERRmine (1.1.2) - Latest version of endorS.py (0.4) -## [2.1.0] - 2020-03-05 - "Ravensburg" +## [2.1.0] - Ravensburg - 2020-03-05 ### `Added` From 9f8240357d0f125d2b4267b3076afb58e312fda8 Mon Sep 17 00:00:00 2001 From: "Thiseas C. Lamnidis" Date: Thu, 18 Nov 2021 10:17:01 +0100 Subject: [PATCH 03/25] split bam_trim options by library strandedness --- nextflow_schema.json | 52 ++++++++++++++++++++++++++++++++++---------- 1 file changed, 40 insertions(+), 12 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 60629e272..31e542802 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -964,33 +964,61 @@ "fa_icon": "fas fa-power-off", "help_text": "Turns on the BAM trimming method. Trims off `[n]` bases from reads in the deduplicated BAM file. Damage assessment in PMDTools or DamageProfiler remains untouched, as data is routed through this independently. BAM trimming is typically performed to reduce errors during genotyping that can be caused by aDNA damage.\n\nBAM trimming will only be performed on libraries indicated as `--udg_type 'none'` or `--udg_type 'half'`. Complete UDG treatment ('full') should have removed all damage. The amount of bases that will be trimmed off can be set separately for libraries with `--udg_type` `'none'` and `'half'` (see `--bamutils_clip_half_udg_left` / `--bamutils_clip_half_udg_right` / `--bamutils_clip_none_udg_left` / `--bamutils_clip_none_udg_right`).\n\n> Note: additional artefacts such as bar-codes or adapters that could potentially also be trimmed should be removed prior mapping." }, - "bamutils_clip_half_udg_left": { + "bamutils_clip_double_stranded_half_udg_left": { "type": "integer", "default": 0, "fa_icon": "fas fa-ruler-combined", - "description": "Specify the number of bases to clip off reads from 'left' end of read for half-UDG libraries.", - "help_text": "Default set to `1` and clips off one base of the left or right side of reads from libraries whose UDG treatment is set to `half`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" + "description": "Specify the number of bases to clip off reads from 'left' end of read for double-stranded half-UDG libraries.", + "help_text": "Default set to `0` and clips off no bases on the left or right side of reads from double_stranded libraries whose UDG treatment is set to `half`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" }, - "bamutils_clip_half_udg_right": { + "bamutils_clip_double_stranded_half_udg_right": { "type": "integer", "default": 0, "fa_icon": "fas fa-ruler", - "description": "Specify the number of bases to clip off reads from 'right' end of read for half-UDG libraries.", - "help_text": "Default set to `1` and clips off one base of the left or right side of reads from libraries whose UDG treatment is set to `half`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" + "description": "Specify the number of bases to clip off reads from 'right' end of read for double-stranded half-UDG libraries.", + "help_text": "Default set to `0` and clips off no bases on the left or right side of reads from double_stranded libraries whose UDG treatment is set to `half`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" }, - "bamutils_clip_none_udg_left": { + "bamutils_clip_double_stranded_none_udg_left": { "type": "integer", "default": 0, "fa_icon": "fas fa-ruler-combined", - "description": "Specify the number of bases to clip off reads from 'left' end of read for non-UDG libraries.", - "help_text": "Default set to `1` and clips off one base of the left or right side of reads from libraries whose UDG treatment is set to `none`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" + "description": "Specify the number of bases to clip off reads from 'left' end of read for double-stranded non-UDG libraries.", + "help_text": "Default set to `0` and clips off no bases on the left or right side of reads from double_stranded libraries whose UDG treatment is set to `none`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" }, - "bamutils_clip_none_udg_right": { + "bamutils_clip_double_stranded_none_udg_right": { "type": "integer", "default": 0, "fa_icon": "fas fa-ruler", - "description": "Specify the number of bases to clip off reads from 'right' end of read for non-UDG libraries.", - "help_text": "Default set to `1` and clips off one base of the left or right side of reads from libraries whose UDG treatment is set to `none`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" + "description": "Specify the number of bases to clip off reads from 'right' end of read for double-stranded non-UDG libraries.", + "help_text": "Default set to `0` and clips off no bases on the left or right side of reads from double_stranded libraries whose UDG treatment is set to `none`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" + }, + "bamutils_clip_single_stranded_half_udg_left": { + "type": "integer", + "default": 0, + "fa_icon": "fas fa-ruler-combined", + "description": "Specify the number of bases to clip off reads from 'left' end of read for single-stranded half-UDG libraries.", + "help_text": "Default set to `0` and clips off no bases on the left or right side of reads from single-stranded libraries whose UDG treatment is set to `half`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" + }, + "bamutils_clip_single_stranded_half_udg_right": { + "type": "integer", + "default": 0, + "fa_icon": "fas fa-ruler", + "description": "Specify the number of bases to clip off reads from 'right' end of read for single-stranded half-UDG libraries.", + "help_text": "Default set to `0` and clips off no bases on the left or right side of reads from single-stranded libraries whose UDG treatment is set to `half`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" + }, + "bamutils_clip_single_stranded_none_udg_left": { + "type": "integer", + "default": 0, + "fa_icon": "fas fa-ruler-combined", + "description": "Specify the number of bases to clip off reads from 'left' end of read for single-stranded non-UDG libraries.", + "help_text": "Default set to `0` and clips off no bases on the left or right side of reads from single-stranded libraries whose UDG treatment is set to `none`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" + }, + "bamutils_clip_single_stranded_none_udg_right": { + "type": "integer", + "default": 0, + "fa_icon": "fas fa-ruler", + "description": "Specify the number of bases to clip off reads from 'right' end of read for single-stranded non-UDG libraries.", + "help_text": "Default set to `0` and clips off no bases on the left or right side of reads from single-stranded libraries whose UDG treatment is set to `none`. Note that reverse reads will automatically be clipped off at the reverse side with this (automatically reverses left and right for the reverse read).\n\n> Modifies bamUtil's trimBam parameter: `-L -R`" }, "bamutils_softclip": { "type": "boolean", From d50d214e2037d9e357fdbcdce31882a5f73ca8ff Mon Sep 17 00:00:00 2001 From: "Thiseas C. Lamnidis" Date: Thu, 18 Nov 2021 10:17:14 +0100 Subject: [PATCH 04/25] Flexible trimming by library strandedness --- main.nf | 7 +++++-- 1 file changed, 5 insertions(+), 2 deletions(-) diff --git a/main.nf b/main.nf index b76f3da16..75b9031f5 100644 --- a/main.nf +++ b/main.nf @@ -2204,8 +2204,11 @@ process bam_trim { script: def softclip = params.bamutils_softclip ? '-c' : '' def size = params.large_ref ? '-c' : '' - def left_clipping = udg == "half" ? "${params.bamutils_clip_half_udg_left}" : "${params.bamutils_clip_none_udg_left}" - def right_clipping = udg == "half" ? "${params.bamutils_clip_half_udg_right}" : "${params.bamutils_clip_none_udg_right}" + def left_clipping = strandedness == "double" ? (udg == "half" ? "${params.bamutils_clip_double_stranded_half_udg_left}" : "${params.bamutils_clip_double_stranded_none_udg_left}") : (udg == "half" ? "${params.bamutils_clip_single_stranded_half_udg_left}" : "${params.bamutils_clip_single_stranded_none_udg_left}") + def right_clipping = strandedness == "double" ? (udg == "half" ? "${params.bamutils_clip_double_stranded_half_udg_right}" : "${params.bamutils_clip_double_stranded_none_udg_right}") : (udg == "half" ? "${params.bamutils_clip_single_stranded_half_udg_right}" : "${params.bamutils_clip_single_stranded_none_udg_right}") + + // def left_clipping = udg == "half" ? "${params.bamutils_clip_half_udg_left}" : "${params.bamutils_clip_none_udg_left}" + // def right_clipping = udg == "half" ? "${params.bamutils_clip_half_udg_right}" : "${params.bamutils_clip_none_udg_right}" """ bam trimBam $bam tmp.bam -L ${left_clipping} -R ${right_clipping} ${softclip} samtools sort -@ ${task.cpus} tmp.bam -o ${libraryid}.trimmed.bam From eddc97e5fa9fcd38524674215debb03c269c1134 Mon Sep 17 00:00:00 2001 From: "Thiseas C. Lamnidis" Date: Thu, 18 Nov 2021 10:36:02 +0100 Subject: [PATCH 05/25] Added new params for bam_trim --- nextflow.config | 12 ++++++++---- nextflow_schema.json | 2 +- 2 files changed, 9 insertions(+), 5 deletions(-) diff --git a/nextflow.config b/nextflow.config index a1952c731..44a0d0e72 100644 --- a/nextflow.config +++ b/nextflow.config @@ -144,10 +144,14 @@ params { //bamUtils trimbam settings run_trim_bam = false - bamutils_clip_half_udg_left = 0 - bamutils_clip_half_udg_right = 0 - bamutils_clip_none_udg_left = 0 - bamutils_clip_none_udg_right = 0 + bamutils_clip_double_stranded_half_udg_left = 0 + bamutils_clip_double_stranded_half_udg_right = 0 + bamutils_clip_double_stranded_none_udg_left = 0 + bamutils_clip_double_stranded_none_udg_right = 0 + bamutils_clip_single_stranded_half_udg_left = 0 + bamutils_clip_single_stranded_half_udg_right = 0 + bamutils_clip_single_stranded_none_udg_left = 0 + bamutils_clip_single_stranded_none_udg_right = 0 bamutils_softclip = false //Genotyping options diff --git a/nextflow_schema.json b/nextflow_schema.json index 31e542802..c140dbe79 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -1764,4 +1764,4 @@ "$ref": "#/definitions/metagenomic_authentication" } ] -} +} \ No newline at end of file From 47cb4152b040bb82918e02e86126419e5ad89494 Mon Sep 17 00:00:00 2001 From: "Thiseas C. Lamnidis" Date: Thu, 18 Nov 2021 10:50:59 +0100 Subject: [PATCH 06/25] Update CHANGELOG.md --- CHANGELOG.md | 6 ++++++ 1 file changed, 6 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index bf8c396b0..dc2f6653c 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,6 +3,12 @@ The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). +## [2.4.1] - 2021-11-18 + +### `Added` + +- [#805](https://github.com/nf-core/eager/issues/805) Changes to bam_trim options to allow flexible trimming by library strandedness (in addition to UDG treatment). + ## [2.4.0] - 20201-09-14 ### `Added` From 13523eebe9161ad261174bdb2ce59ee0c7d9169f Mon Sep 17 00:00:00 2001 From: "Thiseas C. Lamnidis" Date: Thu, 18 Nov 2021 10:51:17 +0100 Subject: [PATCH 07/25] Cerrected release data of 2.4.0 --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index dc2f6653c..5e1e27db7 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -9,7 +9,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. - [#805](https://github.com/nf-core/eager/issues/805) Changes to bam_trim options to allow flexible trimming by library strandedness (in addition to UDG treatment). -## [2.4.0] - 20201-09-14 +## [2.4.0] - 2021-09-14 ### `Added` From ef024b5d4b0d0a7183b455253f377dbc43025785 Mon Sep 17 00:00:00 2001 From: Thiseas Christos Lamnidis Date: Thu, 18 Nov 2021 11:01:06 +0100 Subject: [PATCH 08/25] lint --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 74faa52a5..a8de0aa86 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -180,7 +180,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Deprecated` -## [2.3.0] - Aalen - 2021-01-11 +## [2.3.0] - Aalen - 2021-01-11 ### `Added` From 21a9b1f8bef235501f2f9ef0dff127cd32cdae6d Mon Sep 17 00:00:00 2001 From: Thiseas Christos Lamnidis Date: Thu, 18 Nov 2021 16:11:31 +0100 Subject: [PATCH 09/25] Sample name as SM in bam read groups. Retain RG info on bam merges --- main.nf | 22 +++++++++++----------- 1 file changed, 11 insertions(+), 11 deletions(-) diff --git a/main.nf b/main.nf index b76f3da16..36d36a592 100644 --- a/main.nf +++ b/main.nf @@ -1271,14 +1271,14 @@ process bwa { """ bwa aln -t ${task.cpus} $fasta ${r1} -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -o ${params.bwaalno} -f ${libraryid}.r1.sai bwa aln -t ${task.cpus} $fasta ${r2} -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -o ${params.bwaalno} -f ${libraryid}.r2.sai - bwa sampe -r "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${libraryid}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" $fasta ${libraryid}.r1.sai ${libraryid}.r2.sai ${r1} ${r2} | samtools sort -@ ${task.cpus - 1} -O bam - > ${libraryid}_"${seqtype}".mapped.bam + bwa sampe -r "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${samplename}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" $fasta ${libraryid}.r1.sai ${libraryid}.r2.sai ${r1} ${r2} | samtools sort -@ ${task.cpus - 1} -O bam - > ${libraryid}_"${seqtype}".mapped.bam samtools index "${libraryid}"_"${seqtype}".mapped.bam ${size} """ } else { //PE collapsed, or SE data """ bwa aln -t ${task.cpus} ${fasta} ${r1} -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -o ${params.bwaalno} -f ${libraryid}.sai - bwa samse -r "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${libraryid}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" $fasta ${libraryid}.sai $r1 | samtools sort -@ ${task.cpus - 1} -O bam - > "${libraryid}"_"${seqtype}".mapped.bam + bwa samse -r "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${samplename}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" $fasta ${libraryid}.sai $r1 | samtools sort -@ ${task.cpus - 1} -O bam - > "${libraryid}"_"${seqtype}".mapped.bam samtools index "${libraryid}"_"${seqtype}".mapped.bam ${size} """ } @@ -1309,12 +1309,12 @@ process bwamem { if (!params.single_end && params.skip_collapse){ """ - bwa mem -t ${split_cpus} $fasta $r1 $r2 -R "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${libraryid}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" | samtools sort -@ ${split_cpus} -O bam - > "${libraryid}"_"${seqtype}".mapped.bam + bwa mem -t ${split_cpus} $fasta $r1 $r2 -R "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${samplename}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" | samtools sort -@ ${split_cpus} -O bam - > "${libraryid}"_"${seqtype}".mapped.bam samtools index ${size} -@ ${task.cpus} "${libraryid}".mapped.bam """ } else { """ - bwa mem -t ${split_cpus} $fasta $r1 -R "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${libraryid}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" | samtools sort -@ ${split_cpus} -O bam - > "${libraryid}"_"${seqtype}".mapped.bam + bwa mem -t ${split_cpus} $fasta $r1 -R "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${samplename}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" | samtools sort -@ ${split_cpus} -O bam - > "${libraryid}"_"${seqtype}".mapped.bam samtools index -@ ${task.cpus} "${libraryid}"_"${seqtype}".mapped.bam ${size} """ } @@ -1378,7 +1378,7 @@ process circularmapper{ """ bwa aln -t ${task.cpus} $elongated_root $r1 -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f ${libraryid}.r1.sai bwa aln -t ${task.cpus} $elongated_root $r2 -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f ${libraryid}.r2.sai - bwa sampe -r "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${libraryid}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" $elongated_root ${libraryid}.r1.sai ${libraryid}.r2.sai $r1 $r2 > tmp.out + bwa sampe -r "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${samplename}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" $elongated_root ${libraryid}.r1.sai ${libraryid}.r2.sai $r1 $r2 > tmp.out realignsamfile -Xmx${task.memory.toGiga()}g -e ${params.circularextension} -i tmp.out -r $fasta $filter samtools sort -@ ${task.cpus} -O bam tmp_realigned.bam > ${libraryid}_"${seqtype}".mapped.bam samtools index "${libraryid}"_"${seqtype}".mapped.bam ${size} @@ -1386,7 +1386,7 @@ process circularmapper{ } else { """ bwa aln -t ${task.cpus} $elongated_root $r1 -n ${params.bwaalnn} -l ${params.bwaalnl} -k ${params.bwaalnk} -f ${libraryid}.sai - bwa samse -r "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${libraryid}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" $elongated_root ${libraryid}.sai $r1 > tmp.out + bwa samse -r "@RG\\tID:ILLUMINA-${libraryid}\\tSM:${samplename}\\tPL:illumina\\tPU:ILLUMINA-${libraryid}-${seqtype}" $elongated_root ${libraryid}.sai $r1 > tmp.out realignsamfile -Xmx${task.memory.toGiga()}g -e ${params.circularextension} -i tmp.out -r $fasta $filter samtools sort -@ ${task.cpus} -O bam tmp_realigned.bam > "${libraryid}"_"${seqtype}".mapped.bam samtools index "${libraryid}"_"${seqtype}".mapped.bam ${size} @@ -1456,13 +1456,13 @@ process bowtie2 { //PE data without merging, PE data without any AR applied if ( seqtype == 'PE' && ( params.skip_collapse || params.skip_adapterremoval ) ){ """ - bowtie2 -x ${fasta} -1 ${r1} -2 ${r2} -p ${split_cpus} ${sensitivity} ${bt2n} ${bt2l} ${trim5} ${trim3} --maxins ${params.bt2_maxins} --rg-id ILLUMINA-${libraryid} --rg SM:${libraryid} --rg PL:illumina --rg PU:ILLUMINA-${libraryid}-${seqtype} 2> "${libraryid}"_bt2.log | samtools sort -@ ${split_cpus} -O bam > "${libraryid}"_"${seqtype}".mapped.bam + bowtie2 -x ${fasta} -1 ${r1} -2 ${r2} -p ${split_cpus} ${sensitivity} ${bt2n} ${bt2l} ${trim5} ${trim3} --maxins ${params.bt2_maxins} --rg-id ILLUMINA-${libraryid} --rg SM:${samplename} --rg PL:illumina --rg PU:ILLUMINA-${libraryid}-${seqtype} 2> "${libraryid}"_bt2.log | samtools sort -@ ${split_cpus} -O bam > "${libraryid}"_"${seqtype}".mapped.bam samtools index "${libraryid}"_"${seqtype}".mapped.bam ${size} """ } else { //PE collapsed, or SE data """ - bowtie2 -x ${fasta} -U ${r1} -p ${split_cpus} ${sensitivity} ${bt2n} ${bt2l} ${trim5} ${trim3} --rg-id ILLUMINA-${libraryid} --rg SM:${libraryid} --rg PL:illumina --rg PU:ILLUMINA-${libraryid}-${seqtype} 2> "${libraryid}"_bt2.log | samtools sort -@ ${split_cpus} -O bam > "${libraryid}"_"${seqtype}".mapped.bam + bowtie2 -x ${fasta} -U ${r1} -p ${split_cpus} ${sensitivity} ${bt2n} ${bt2l} ${trim5} ${trim3} --rg-id ILLUMINA-${libraryid} --rg SM:${samplename} --rg PL:illumina --rg PU:ILLUMINA-${libraryid}-${seqtype} 2> "${libraryid}"_bt2.log | samtools sort -@ ${split_cpus} -O bam > "${libraryid}"_"${seqtype}".mapped.bam samtools index "${libraryid}"_"${seqtype}".mapped.bam ${size} """ } @@ -1594,7 +1594,7 @@ ch_branched_for_seqtypemerge = ch_mapping_for_seqtype_merging """ samtools merge ${libraryid}_seqtypemerged.bam ${bam} ## Have to set validation as lenient because of BWA issue: "I see a read stands out the end of a chromosome and is flagged as unmapped (flag 0x4). [...]" http://bio-bwa.sourceforge.net/ - picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${libraryid}_seqtypemerged.bam O=${libraryid}_seqtypemerged_rg.bam RGID=1 RGLB="${libraryid}_seqtypemerged" RGPL=illumina RGPU=4410 RGSM="${libraryid}_seqtypemerged" VALIDATION_STRINGENCY=LENIENT + # picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${libraryid}_seqtypemerged.bam O=${libraryid}_seqtypemerged_rg.bam RGID=1 RGLB="${libraryid}_seqtypemerged" RGPL=illumina RGPU=4410 RGSM="${libraryid}_seqtypemerged" VALIDATION_STRINGENCY=LENIENT samtools index ${libraryid}_seqtypemerged_rg.bam ${size} """ @@ -1965,7 +1965,7 @@ process library_merge { """ samtools merge ${samplename}_libmerged_rmdup.bam ${bam} ## Have to set validation as lenient because of BWA issue: "I see a read stands out the end of a chromosome and is flagged as unmapped (flag 0x4). [...]" http://bio-bwa.sourceforge.net/ - picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${samplename}_libmerged_rmdup.bam O=${samplename}_libmerged_rg_rmdup.bam RGID=1 RGLB="${samplename}_merged" RGPL=illumina RGPU=4410 RGSM="${samplename}_merged" VALIDATION_STRINGENCY=LENIENT + # picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${samplename}_libmerged_rmdup.bam O=${samplename}_libmerged_rg_rmdup.bam RGID=1 RGLB="${samplename}_merged" RGPL=illumina RGPU=4410 RGSM="${samplename}_merged" VALIDATION_STRINGENCY=LENIENT samtools index ${samplename}_libmerged_rg_rmdup.bam ${size} """ } @@ -2258,7 +2258,7 @@ process additional_library_merge { def size = params.large_ref ? '-c' : '' """ samtools merge ${samplename}_libmerged_add.bam ${bam} - picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${samplename}_libmerged_add.bam O=${samplename}_libmerged_rg_add.bam RGID=1 RGLB="${samplename}_additionalmerged" RGPL=illumina RGPU=4410 RGSM="${samplename}_additionalmerged" VALIDATION_STRINGENCY=LENIENT + # picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${samplename}_libmerged_add.bam O=${samplename}_libmerged_rg_add.bam RGID=1 RGLB="${samplename}_additionalmerged" RGPL=illumina RGPU=4410 RGSM="${samplename}_additionalmerged" VALIDATION_STRINGENCY=LENIENT samtools index ${samplename}_libmerged_rg_add.bam ${size} """ } From ef525d178a62b25e11aca1b7ae579897bc961b5d Mon Sep 17 00:00:00 2001 From: Thiseas Christos Lamnidis Date: Thu, 18 Nov 2021 16:32:44 +0100 Subject: [PATCH 10/25] Update CHANGELOG.md --- CHANGELOG.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index dbbc3ce01..b3320766f 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -7,6 +7,8 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` + - [#808](https://github.com/nf-core/eager/issues/808) Retain read group information across bam merges. Sample set to Sample name in bwa output RG. + ### `Fixed` ### `Dependencies` From 79fedba361a4c6549f61d24abeeab93c360667a6 Mon Sep 17 00:00:00 2001 From: Thiseas Christos Lamnidis Date: Thu, 18 Nov 2021 16:50:45 +0100 Subject: [PATCH 11/25] Flexible filtering of reads for pileupcaller genotyping. --- CHANGELOG.md | 1 + main.nf | 7 +++++-- nextflow.config | 2 ++ nextflow_schema.json | 18 ++++++++++++++++-- 4 files changed, 24 insertions(+), 4 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index b3320766f..fc5f556c4 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -8,6 +8,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` - [#808](https://github.com/nf-core/eager/issues/808) Retain read group information across bam merges. Sample set to Sample name in bwa output RG. + - Map and base quality filters prior to genotyping with pileupcaller can now be specified. ### `Fixed` diff --git a/main.nf b/main.nf index 36d36a592..42aa09f88 100644 --- a/main.nf +++ b/main.nf @@ -2488,10 +2488,13 @@ process genotyping_pileupcaller { def ssmode = strandedness == "single" ? "--singleStrandMode" : "" def bam_list = bam.flatten().join(" ") def sample_names = samplename.flatten().join(",") + def map_q = params.pileupcaller_min_map_quality + def base_q = params.pileupcaller_min_base_quality + """ - samtools mpileup -B -q 30 -Q 30 ${use_bed} -f ${fasta} ${bam_list} | pileupCaller ${caller} ${ssmode} ${transitions_mode} --sampleNames ${sample_names} ${use_snp} -e pileupcaller.${strandedness} + samtools mpileup -B --ignore-RG -q ${map_q} -Q ${base_q} ${use_bed} -f ${fasta} ${bam_list} | pileupCaller ${caller} ${ssmode} ${transitions_mode} --sampleNames ${sample_names} ${use_snp} -e pileupcaller.${strandedness} """ -} +}} process eigenstrat_snp_coverage { label 'mc_tiny' diff --git a/nextflow.config b/nextflow.config index f86b9dbd8..df37dd6e4 100644 --- a/nextflow.config +++ b/nextflow.config @@ -174,6 +174,8 @@ params { pileupcaller_bedfile = null pileupcaller_method = 'randomHaploid' pileupcaller_transitions_mode = 'AllSites' + pileupcaller_min_map_quality = 30 + pileupcaller_min_base_quality = 30 // ANGSD Genotype Likelihoods angsd_glmodel = 'samtools' angsd_glformat = 'binary' diff --git a/nextflow_schema.json b/nextflow_schema.json index 60629e272..ae2f7e442 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -1185,6 +1185,20 @@ "SkipTransitions" ] }, + "pileupcaller_min_map_quality": { + "type": "integer", + "default": 30, + "description": "The minimum mapping quality to be used for genotyping.", + "fa_icon": "fas fa-filter", + "help_text": "The minimum mapping quality to be used for genotyping. Affects the `samtools pileup` output that is used by pileupcaller. Affects `-q` parameter of samtools mpileup." + }, + "pileupcaller_min_base_quality": { + "type": "integer", + "default": 30, + "description": "The minimum base quality to be used for genotyping.", + "fa_icon": "fas fa-filter", + "help_text": "The minimum base quality to be used for genotyping. Affects the `samtools pileup` output that is used by pileupcaller. Affects `-Q` parameter of samtools mpileup." + }, "angsd_glmodel": { "type": "string", "default": "samtools", @@ -1641,7 +1655,7 @@ "maltextract_percentidentity": { "type": "number", "description": "Minimum percent identity alignments are required to have to be reported. Recommended to set same as MALT parameter.", - "default": 85, + "default": 85.0, "fa_icon": "fas fa-id-card", "help_text": "Minimum percent identity alignments are required to have to be reported. Higher values allows fewer mismatches between read and reference sequence, but therefore will provide greater confidence in the hit. Lower values allow more mismatches, which can account for damage and divergence of a related strain/species to the reference. Recommended to set same as MALT parameter or higher. Default: `85.0`.\n\nOnly when `--metagenomic_tool malt` is also supplied.\n\n> Modifies MaltExtract parameter: `--minPI`" }, @@ -1736,4 +1750,4 @@ "$ref": "#/definitions/metagenomic_authentication" } ] -} +} \ No newline at end of file From 0344f01f236ca32ad158fb8452a50fa5f8da612e Mon Sep 17 00:00:00 2001 From: Thiseas Christos Lamnidis Date: Thu, 18 Nov 2021 16:56:46 +0100 Subject: [PATCH 12/25] Fix typo --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index 42aa09f88..9de4fac9f 100644 --- a/main.nf +++ b/main.nf @@ -2494,7 +2494,7 @@ process genotyping_pileupcaller { """ samtools mpileup -B --ignore-RG -q ${map_q} -Q ${base_q} ${use_bed} -f ${fasta} ${bam_list} | pileupCaller ${caller} ${ssmode} ${transitions_mode} --sampleNames ${sample_names} ${use_snp} -e pileupcaller.${strandedness} """ -}} +} process eigenstrat_snp_coverage { label 'mc_tiny' From 6cf64b73e03cd4b73c592b92e7b59558b6a8168f Mon Sep 17 00:00:00 2001 From: "Thiseas C. Lamnidis" Date: Thu, 18 Nov 2021 19:04:37 +0100 Subject: [PATCH 13/25] Correct bam merging output name --- main.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/main.nf b/main.nf index 36d36a592..e0fc88469 100644 --- a/main.nf +++ b/main.nf @@ -1595,7 +1595,7 @@ ch_branched_for_seqtypemerge = ch_mapping_for_seqtype_merging samtools merge ${libraryid}_seqtypemerged.bam ${bam} ## Have to set validation as lenient because of BWA issue: "I see a read stands out the end of a chromosome and is flagged as unmapped (flag 0x4). [...]" http://bio-bwa.sourceforge.net/ # picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${libraryid}_seqtypemerged.bam O=${libraryid}_seqtypemerged_rg.bam RGID=1 RGLB="${libraryid}_seqtypemerged" RGPL=illumina RGPU=4410 RGSM="${libraryid}_seqtypemerged" VALIDATION_STRINGENCY=LENIENT - samtools index ${libraryid}_seqtypemerged_rg.bam ${size} + samtools index ${libraryid}_seqtypemerged.bam ${size} """ } @@ -1966,7 +1966,7 @@ process library_merge { samtools merge ${samplename}_libmerged_rmdup.bam ${bam} ## Have to set validation as lenient because of BWA issue: "I see a read stands out the end of a chromosome and is flagged as unmapped (flag 0x4). [...]" http://bio-bwa.sourceforge.net/ # picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${samplename}_libmerged_rmdup.bam O=${samplename}_libmerged_rg_rmdup.bam RGID=1 RGLB="${samplename}_merged" RGPL=illumina RGPU=4410 RGSM="${samplename}_merged" VALIDATION_STRINGENCY=LENIENT - samtools index ${samplename}_libmerged_rg_rmdup.bam ${size} + samtools index ${samplename}_libmerged_rmdup.bam ${size} """ } @@ -2259,7 +2259,7 @@ process additional_library_merge { """ samtools merge ${samplename}_libmerged_add.bam ${bam} # picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${samplename}_libmerged_add.bam O=${samplename}_libmerged_rg_add.bam RGID=1 RGLB="${samplename}_additionalmerged" RGPL=illumina RGPU=4410 RGSM="${samplename}_additionalmerged" VALIDATION_STRINGENCY=LENIENT - samtools index ${samplename}_libmerged_rg_add.bam ${size} + samtools index ${samplename}_libmerged_add.bam ${size} """ } From 5bbcd3083092e3b8f9a9a34663328d17631f68d1 Mon Sep 17 00:00:00 2001 From: "Thiseas C. Lamnidis" Date: Thu, 18 Nov 2021 20:41:04 +0100 Subject: [PATCH 14/25] Correct process output for bam merging proceses --- main.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/main.nf b/main.nf index e0fc88469..6d0df1873 100644 --- a/main.nf +++ b/main.nf @@ -1587,7 +1587,7 @@ ch_branched_for_seqtypemerge = ch_mapping_for_seqtype_merging tuple samplename, libraryid, lane, seqtype, organism, strandedness, udg, file(bam), file(bai) from ch_branched_for_seqtypemerge.merge_me output: - tuple samplename, libraryid, lane, seqtype, organism, strandedness, udg, file("*_seqtypemerged_rg.bam"), file("*_seqtypemerged_rg*.{bai,csi}") into ch_seqtypemerge_for_filtering + tuple samplename, libraryid, lane, seqtype, organism, strandedness, udg, file("*_seqtypemerged.bam"), file("*_seqtypemerged*.{bai,csi}") into ch_seqtypemerge_for_filtering script: def size = params.large_ref ? '-c' : '' @@ -1958,7 +1958,7 @@ process library_merge { tuple samplename, libraryid, lane, seqtype, organism, strandedness, udg, file(bam), file(bai) from ch_fixedinput_for_librarymerging.dump(tag: "library_merge_input") output: - tuple samplename, val("${samplename}_libmerged"), lane, seqtype, organism, strandedness, udg, path("*_libmerged_rg_rmdup.bam"), path("*_libmerged_rg_rmdup.bam.{bai,csi}") into ch_output_from_librarymerging + tuple samplename, val("${samplename}_libmerged"), lane, seqtype, organism, strandedness, udg, path("*_libmerged_rmdup.bam"), path("*_libmerged_rmdup.bam.{bai,csi}") into ch_output_from_librarymerging script: def size = params.large_ref ? '-c' : '' @@ -2252,7 +2252,7 @@ process additional_library_merge { tuple samplename, libraryid, lane, seqtype, organism, strandedness, udg, path(bam), path(bai) from ch_trimmed_formerge.merge_me output: - tuple samplename, val("${samplename}_libmerged"), lane, seqtype, organism, strandedness, udg, path("*_libmerged_rg_add.bam"), path("*_libmerged_rg_add.bam.{bai,csi}") into ch_output_from_trimmerge + tuple samplename, val("${samplename}_libmerged"), lane, seqtype, organism, strandedness, udg, path("*_libmerged_add.bam"), path("*_libmerged_add.bam.{bai,csi}") into ch_output_from_trimmerge script: def size = params.large_ref ? '-c' : '' From 2db00fb79603beec498773e5577792aaa4c422a6 Mon Sep 17 00:00:00 2001 From: "Thiseas C. Lamnidis" Date: Fri, 19 Nov 2021 19:17:42 +0100 Subject: [PATCH 15/25] Update CHANGELOG.md --- CHANGELOG.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index fc5f556c4..9a7b530da 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -7,8 +7,8 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` - - [#808](https://github.com/nf-core/eager/issues/808) Retain read group information across bam merges. Sample set to Sample name in bwa output RG. - - Map and base quality filters prior to genotyping with pileupcaller can now be specified. +- [#808](https://github.com/nf-core/eager/issues/808) Retain read group information across bam merges. Sample set to Sample name in bwa output RG. +- Map and base quality filters prior to genotyping with pileupcaller can now be specified. ### `Fixed` From b5802f9b01bc0f74e65e479a291996569c196fd6 Mon Sep 17 00:00:00 2001 From: "James A. Fellows Yates" Date: Tue, 23 Nov 2021 20:24:17 +0100 Subject: [PATCH 16/25] Update CHANGELOG.md --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 9a7b530da..dd3c68a3d 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -181,7 +181,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Deprecated` -## [2.3.0] - Aalen - 2021-01-11 +## [2.3.0] - Aalen - 2021-01-11 ### `Added` From fb124c27ce5e6829d56e9e86193e4d3f4ff0dcdf Mon Sep 17 00:00:00 2001 From: "Thiseas C. Lamnidis" Date: Tue, 23 Nov 2021 21:19:53 +0100 Subject: [PATCH 17/25] Update CHANGELOG.md Co-authored-by: James A. Fellows Yates --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index dd3c68a3d..c3e6a3000 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -7,7 +7,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` -- [#808](https://github.com/nf-core/eager/issues/808) Retain read group information across bam merges. Sample set to Sample name in bwa output RG. +- [#808](https://github.com/nf-core/eager/issues/808) Retain read group information across bam merges. Sample set to sample name (rather than library name) in bwa output 'RG' readgroup tag. - Map and base quality filters prior to genotyping with pileupcaller can now be specified. ### `Fixed` From cecbf0b018e889dfa2046da0b2828455a3541cea Mon Sep 17 00:00:00 2001 From: "Thiseas C. Lamnidis" Date: Tue, 23 Nov 2021 21:22:19 +0100 Subject: [PATCH 18/25] Removed commented old code. --- main.nf | 5 ----- 1 file changed, 5 deletions(-) diff --git a/main.nf b/main.nf index e7610ef7f..5292e3a8a 100644 --- a/main.nf +++ b/main.nf @@ -1593,8 +1593,6 @@ ch_branched_for_seqtypemerge = ch_mapping_for_seqtype_merging def size = params.large_ref ? '-c' : '' """ samtools merge ${libraryid}_seqtypemerged.bam ${bam} - ## Have to set validation as lenient because of BWA issue: "I see a read stands out the end of a chromosome and is flagged as unmapped (flag 0x4). [...]" http://bio-bwa.sourceforge.net/ - # picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${libraryid}_seqtypemerged.bam O=${libraryid}_seqtypemerged_rg.bam RGID=1 RGLB="${libraryid}_seqtypemerged" RGPL=illumina RGPU=4410 RGSM="${libraryid}_seqtypemerged" VALIDATION_STRINGENCY=LENIENT samtools index ${libraryid}_seqtypemerged.bam ${size} """ @@ -1964,8 +1962,6 @@ process library_merge { def size = params.large_ref ? '-c' : '' """ samtools merge ${samplename}_libmerged_rmdup.bam ${bam} - ## Have to set validation as lenient because of BWA issue: "I see a read stands out the end of a chromosome and is flagged as unmapped (flag 0x4). [...]" http://bio-bwa.sourceforge.net/ - # picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${samplename}_libmerged_rmdup.bam O=${samplename}_libmerged_rg_rmdup.bam RGID=1 RGLB="${samplename}_merged" RGPL=illumina RGPU=4410 RGSM="${samplename}_merged" VALIDATION_STRINGENCY=LENIENT samtools index ${samplename}_libmerged_rmdup.bam ${size} """ } @@ -2258,7 +2254,6 @@ process additional_library_merge { def size = params.large_ref ? '-c' : '' """ samtools merge ${samplename}_libmerged_add.bam ${bam} - # picard -Xmx${task.memory.toGiga()}g AddOrReplaceReadGroups I=${samplename}_libmerged_add.bam O=${samplename}_libmerged_rg_add.bam RGID=1 RGLB="${samplename}_additionalmerged" RGPL=illumina RGPU=4410 RGSM="${samplename}_additionalmerged" VALIDATION_STRINGENCY=LENIENT samtools index ${samplename}_libmerged_add.bam ${size} """ } From 53357faff6d5c52129a2cde37ab30cf99573dd9c Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Mon, 29 Nov 2021 10:39:26 +0100 Subject: [PATCH 19/25] Minor QoL updates --- CHANGELOG.md | 6 +++++- docs/images/usage/eager2_metromap_complex.svg | 14 +++++++------- docs/output.md | 4 ++-- main.nf | 4 ++-- 4 files changed, 16 insertions(+), 12 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index ea88da141..595c0132a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,16 +3,20 @@ The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). -## [dev] +## 2.4.1[dev] ### `Added` - [#805](https://github.com/nf-core/eager/issues/805) Changes to bam_trim options to allow flexible trimming by library strandedness (in addition to UDG treatment). - [#808](https://github.com/nf-core/eager/issues/808) Retain read group information across bam merges. Sample set to sample name (rather than library name) in bwa output 'RG' readgroup tag. - Map and base quality filters prior to genotyping with pileupcaller can now be specified. +- [#774](https://github.com/nf-core/eager/issues/774) Added support for multi-threaded Bowtie2 build reference genome indexing +- [#804](https://github.com/nf-core/eager/issues/804) Improved output documentation description to add how 'cluster factor' is calculated ### `Fixed` +- [#803](https://github.com/nf-core/eager/issues/803) Fixed mistake in metro-map diagram (`samtools index` is now correctly `samtools faidx`) + ### `Dependencies` ### `Deprecated` diff --git a/docs/images/usage/eager2_metromap_complex.svg b/docs/images/usage/eager2_metromap_complex.svg index 0dd3c4985..18b2f1e95 100644 --- a/docs/images/usage/eager2_metromap_complex.svg +++ b/docs/images/usage/eager2_metromap_complex.svg @@ -5,7 +5,7 @@ viewBox="0 0 316.84646 170.04657" version="1.1" id="svg8" - inkscape:version="1.1 (1:1.1+202105261517+ce6663b3b7)" + inkscape:version="1.1.1 (1:1.1+202109281949+c3084ef5ed)" sodipodi:docname="eager2_metromap_complex.svg" inkscape:export-filename="/home/jfellows/Documents/git/nf-core/eager/docs/images/usage/eager2_metromap_complex.png" inkscape:export-xdpi="300" @@ -145,9 +145,9 @@ borderopacity="1.0" inkscape:pageopacity="0.0" inkscape:pageshadow="2" - inkscape:zoom="0.3238356" - inkscape:cx="406.07024" - inkscape:cy="-143.59138" + inkscape:zoom="0.4579727" + inkscape:cx="677.98801" + inkscape:cy="506.58042" inkscape:document-units="mm" inkscape:current-layer="layer1" inkscape:document-rotation="0" @@ -155,9 +155,9 @@ inkscape:snap-bbox="true" inkscape:bbox-nodes="true" inkscape:window-width="1920" - inkscape:window-height="1016" + inkscape:window-height="1043" inkscape:window-x="0" - inkscape:window-y="27" + inkscape:window-y="0" inkscape:window-maximized="1" inkscape:snap-bbox-midpoints="false" inkscape:snap-global="true" @@ -967,7 +967,7 @@ x="143.71013" y="116.57104" style="font-style:normal;font-variant:normal;font-weight:normal;font-stretch:normal;font-size:3.52777px;font-family:'Maven Pro';-inkscape-font-specification:'Maven Pro';text-align:center;text-anchor:middle;stroke-width:0.264583" - id="tspan1556">index + id="tspan1556">faidx Z N base** These columns are from DamageProfiler. The prime numbers represent which end of the reads the damage is referring to. The Y>Z is the type of substitution (C>T is the true damage, G>A is the complementary). You should see for no- and half-UDG treatment a decrease in frequency from the 1st to 2nd base. * **Mean Read Length** This is from DamageProfiler. This is the mean length of all de-duplicated mapped reads. Ancient DNA normally will have a mean between 30-75, however this can vary. * **Median Read Length** This is from DamageProfiler. This is the median length of all de-duplicated mapped reads. Ancient DNA normally will have a mean between 30-75, however this can vary. diff --git a/main.nf b/main.nf index 4ffa0e873..804b44609 100644 --- a/main.nf +++ b/main.nf @@ -497,7 +497,7 @@ if( !params.bwa_index && params.fasta && (params.mapper == 'bwaaln' || params.ma // bowtie2 Index if( !params.bt2_index && params.fasta && params.mapper == "bowtie2"){ process makeBT2Index { - label 'sc_medium' + label 'mc_medium' tag "${fasta}" publishDir path: "${params.outdir}/reference_genome/bt2_index", mode: params.publish_dir_mode, saveAs: { filename -> if (params.save_reference) filename @@ -515,7 +515,7 @@ if( !params.bt2_index && params.fasta && params.mapper == "bowtie2"){ script: """ - bowtie2-build $fasta $fasta + bowtie2-build --threads ${task.cpus} $fasta $fasta mkdir BT2Index && mv ${fasta}* BT2Index """ } From becd25667cf0206269011a8c6b8a38229851de64 Mon Sep 17 00:00:00 2001 From: Thiseas Christos Lamnidis Date: Tue, 30 Nov 2021 09:21:39 +0100 Subject: [PATCH 20/25] Verision bump 2.4.1 --- .github/workflows/ci.yml | 4 ++-- CHANGELOG.md | 4 ++-- Dockerfile | 4 ++-- environment.yml | 2 +- nextflow.config | 4 ++-- 5 files changed, 9 insertions(+), 9 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 21b7ae020..70e0ec0ac 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -37,13 +37,13 @@ jobs: - name: Build new docker image if: env.MATCHED_FILES - run: docker build --no-cache . -t nfcore/eager:dev + run: docker build --no-cache . -t nfcore/eager:2.4.1 - name: Pull docker image if: ${{ !env.MATCHED_FILES }} run: | docker pull nfcore/eager:dev - docker tag nfcore/eager:dev nfcore/eager:dev + docker tag nfcore/eager:dev nfcore/eager:2.4.1 - name: Install Nextflow env: diff --git a/CHANGELOG.md b/CHANGELOG.md index 595c0132a..033f389de 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). -## 2.4.1[dev] +## [2.4.1] - 2021-11-30 ### `Added` @@ -21,7 +21,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Deprecated` -## [2.4.0] - Wangen - 20201-09-14 +## [2.4.0] - Wangen - 2021-09-14 ### `Added` diff --git a/Dockerfile b/Dockerfile index 7087c402a..1354408d3 100644 --- a/Dockerfile +++ b/Dockerfile @@ -7,7 +7,7 @@ COPY environment.yml / RUN conda env create --quiet -f /environment.yml && conda clean -a # Add conda installation dir to PATH (instead of doing 'conda activate') -ENV PATH /opt/conda/envs/nf-core-eager-2.4.1dev/bin:$PATH +ENV PATH /opt/conda/envs/nf-core-eager-2.4.1/bin:$PATH # Dump the details of the installed packages to a file for posterity -RUN conda env export --name nf-core-eager-2.4.1dev > nf-core-eager-2.4.1dev.yml \ No newline at end of file +RUN conda env export --name nf-core-eager-2.4.1 > nf-core-eager-2.4.1.yml \ No newline at end of file diff --git a/environment.yml b/environment.yml index 3b007e300..db6c11b10 100644 --- a/environment.yml +++ b/environment.yml @@ -1,6 +1,6 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml -name: nf-core-eager-2.4.1dev +name: nf-core-eager-2.4.1 channels: - conda-forge - bioconda diff --git a/nextflow.config b/nextflow.config index 2ca068747..20076e27e 100644 --- a/nextflow.config +++ b/nextflow.config @@ -284,7 +284,7 @@ params { // Container slug. Stable releases should specify release tag! // Developmental code should specify :dev -process.container = 'nfcore/eager:dev' +process.container = 'nfcore/eager:2.4.1' // Load base.config by default for all pipelines includeConfig 'conf/base.config' @@ -414,7 +414,7 @@ manifest { description = 'A fully reproducible and state-of-the-art ancient DNA analysis pipeline' mainScript = 'main.nf' nextflowVersion = '>=20.07.1' - version = '2.4.1dev' + version = '2.4.1' } // Function to ensure that resource requirements don't go beyond From d2f4be4eab7e823759b73771228b481b2cf092fa Mon Sep 17 00:00:00 2001 From: Thiseas Christos Lamnidis Date: Tue, 30 Nov 2021 09:36:55 +0100 Subject: [PATCH 21/25] Added mentions --- CHANGELOG.md | 12 ++++++------ 1 file changed, 6 insertions(+), 6 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 033f389de..31d4adf35 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -7,15 +7,15 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` -- [#805](https://github.com/nf-core/eager/issues/805) Changes to bam_trim options to allow flexible trimming by library strandedness (in addition to UDG treatment). -- [#808](https://github.com/nf-core/eager/issues/808) Retain read group information across bam merges. Sample set to sample name (rather than library name) in bwa output 'RG' readgroup tag. -- Map and base quality filters prior to genotyping with pileupcaller can now be specified. -- [#774](https://github.com/nf-core/eager/issues/774) Added support for multi-threaded Bowtie2 build reference genome indexing -- [#804](https://github.com/nf-core/eager/issues/804) Improved output documentation description to add how 'cluster factor' is calculated +- [#805](https://github.com/nf-core/eager/issues/805) Changes to bam_trim options to allow flexible trimming by library strandedness (in addition to UDG treatment). (@TCLamnidis) +- [#808](https://github.com/nf-core/eager/issues/808) Retain read group information across bam merges. Sample set to sample name (rather than library name) in bwa output 'RG' readgroup tag. (@TCLamnidis) +- Map and base quality filters prior to genotyping with pileupcaller can now be specified. (@TCLamnidis) +- [#774](https://github.com/nf-core/eager/issues/774) Added support for multi-threaded Bowtie2 build reference genome indexing (@jfy133) +- [#804](https://github.com/nf-core/eager/issues/804) Improved output documentation description to add how 'cluster factor' is calculated (thanks to @meganemichel) ### `Fixed` -- [#803](https://github.com/nf-core/eager/issues/803) Fixed mistake in metro-map diagram (`samtools index` is now correctly `samtools faidx`) +- [#803](https://github.com/nf-core/eager/issues/803) Fixed mistake in metro-map diagram (`samtools index` is now correctly `samtools faidx`) (@jfy133) ### `Dependencies` From b9567ec4b06fb2cc1814c07c90fde43afa2cc6bf Mon Sep 17 00:00:00 2001 From: James Fellows Yates Date: Tue, 30 Nov 2021 09:51:49 +0100 Subject: [PATCH 22/25] Update images --- docs/images/usage/eager2_metromap_complex.png | Bin 549182 -> 549298 bytes docs/images/usage/eager2_metromap_complex.svg | 22 +++++++++--------- 2 files changed, 11 insertions(+), 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