From b7c476c0cbbbe262b61951e42f7def25520c1d9c Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 20 Jun 2022 08:15:36 +0200 Subject: [PATCH 1/8] Added tiddit to pair and tumor VC --- subworkflows/local/pair_variant_calling.nf | 16 +++++++++++-- subworkflows/local/tumor_variant_calling.nf | 17 ++++++++++++-- tests/test_tools.yml | 25 +++++++++++++++++++++ workflows/sarek.nf | 6 +++-- 4 files changed, 58 insertions(+), 6 deletions(-) diff --git a/subworkflows/local/pair_variant_calling.nf b/subworkflows/local/pair_variant_calling.nf index 49d0723d2..c2d25df09 100644 --- a/subworkflows/local/pair_variant_calling.nf +++ b/subworkflows/local/pair_variant_calling.nf @@ -10,6 +10,7 @@ include { RUN_STRELKA_SOMATIC } from '../nf-core/variantca include { RUN_CNVKIT_SOMATIC } from '../nf-core/variantcalling/cnvkit/somatic/main.nf' include { RUN_MPILEUP as RUN_MPILEUP_NORMAL } from '../nf-core/variantcalling/mpileup/main' include { RUN_MPILEUP as RUN_MPILEUP_TUMOR } from '../nf-core/variantcalling/mpileup/main' +include { RUN_TIDDIT } from '../nf-core/variantcalling/tiddit/main.nf' workflow PAIR_VARIANT_CALLING { take: @@ -30,6 +31,7 @@ workflow PAIR_VARIANT_CALLING { panel_of_normals_tbi // channel: [optional] panel_of_normals_tbi chr_files mappability + bwa // channel: [optional] bwa main: @@ -41,6 +43,7 @@ workflow PAIR_VARIANT_CALLING { strelka_vcf = Channel.empty() msisensorpro_output = Channel.empty() mutect2_vcf = Channel.empty() + tiddit_vcf = Channel.empty() // Remap channel with intervals cram_pair_intervals = cram_pair.combine(intervals) @@ -188,8 +191,15 @@ workflow PAIR_VARIANT_CALLING { ch_versions = ch_versions.mix(GATK_TUMOR_NORMAL_SOMATIC_VARIANT_CALLING.out.versions) } - // if (tools.contains('tiddit')) { - // } + //TIDDIT + if (tools.contains('tiddit')){ + RUN_TIDDIT(cram_pair, + fasta, + bwa) + + tiddit_vcf = RUN_TIDDIT.out.tiddit_vcf + ch_versions = ch_versions.mix(RUN_TIDDIT.out.versions) + } emit: freebayes_vcf @@ -197,5 +207,7 @@ workflow PAIR_VARIANT_CALLING { msisensorpro_output mutect2_vcf strelka_vcf + tiddit_vcf + versions = ch_versions } diff --git a/subworkflows/local/tumor_variant_calling.nf b/subworkflows/local/tumor_variant_calling.nf index 0d12f8370..210e93e3e 100644 --- a/subworkflows/local/tumor_variant_calling.nf +++ b/subworkflows/local/tumor_variant_calling.nf @@ -10,6 +10,7 @@ include { RUN_STRELKA_SINGLE } from '../nf-core/variantcall include { RUN_CONTROLFREEC_TUMORONLY } from '../nf-core/variantcalling/controlfreec/tumoronly/main.nf' include { RUN_CNVKIT_TUMORONLY } from '../nf-core/variantcalling/cnvkit/tumoronly/main.nf' include { RUN_MPILEUP } from '../nf-core/variantcalling/mpileup/main' +include { RUN_TIDDIT } from '../nf-core/variantcalling/tiddit/main.nf' workflow TUMOR_ONLY_VARIANT_CALLING { take: @@ -29,6 +30,7 @@ workflow TUMOR_ONLY_VARIANT_CALLING { panel_of_normals_tbi // channel: [optional] panel_of_normals_tbi chr_files mappability + bwa // channel: [optional] bwa main: @@ -39,6 +41,7 @@ workflow TUMOR_ONLY_VARIANT_CALLING { manta_vcf = Channel.empty() mutect2_vcf = Channel.empty() strelka_vcf = Channel.empty() + tiddit_vcf = Channel.empty() // Remap channel with intervals cram_recalibrated_intervals = cram_recalibrated.combine(intervals) @@ -152,16 +155,26 @@ workflow TUMOR_ONLY_VARIANT_CALLING { ch_versions = ch_versions.mix(RUN_STRELKA_SINGLE.out.versions) } + //TIDDIT + if (tools.contains('tiddit')){ + RUN_TIDDIT(cram_recalibrated, + fasta, + bwa) + + tiddit_vcf = RUN_TIDDIT.out.tiddit_vcf + ch_versions = ch_versions.mix(RUN_TIDDIT.out.versions) + } + // if (tools.contains('tiddit')){ // } emit: - versions = ch_versions - freebayes_vcf manta_vcf mutect2_vcf strelka_vcf + tiddit_vcf + versions = ch_versions } diff --git a/tests/test_tools.yml b/tests/test_tools.yml index ab0111ebb..f593f69b5 100644 --- a/tests/test_tools.yml +++ b/tests/test_tools.yml @@ -81,6 +81,31 @@ - path: results/variant_calling/sample1/tiddit/sample1.vcf.gz - path: results/variant_calling/sample1/tiddit/sample1.vcf.gz.tbi +- name: Run variant calling on tumor_only sample with tiddit + command: nextflow run main.nf -profile test,tools_tumoronly,docker --tools tiddit -c ./tests/nextflow.config + tags: + - tiddit + - tumor_only + - variant_calling + files: + - path: results/variant_calling/sample2/tiddit/sample2.ploidies.tab + - path: results/variant_calling/sample2/tiddit/sample2.vcf.gz + - path: results/variant_calling/sample2/tiddit/sample2.vcf.gz.tbi + +- name: Run variant calling on somatic sample with tiddit + command: nextflow run main.nf -profile test,tools_somatic,docker --tools tiddit -c ./tests/nextflow.config + tags: + - tiddit + - somatic + - variant_calling + files: + - path: results/variant_calling/sample4_vs_sample3/tiddit/sample4_vs_sample3.ploidies.tab + - path: results/variant_calling/sample4_vs_sample3/tiddit/sample4_vs_sample3.vcf.gz + - path: results/variant_calling/sample4_vs_sample3/tiddit/sample4_vs_sample3.vcf.gz.tbi + - path: results/variant_calling/sample3/tiddit/sample3.ploidies.tab + - path: results/variant_calling/sample3/tiddit/sample3.vcf.gz + - path: results/variant_calling/sample3/tiddit/sample3.vcf.gz.tbi + - name: Run variant calling on somatic samples with controlfreec command: nextflow run main.nf -profile test,tools_somatic,docker --tools controlfreec -c ./tests/nextflow.config tags: diff --git a/workflows/sarek.nf b/workflows/sarek.nf index 884e9452d..256e72239 100644 --- a/workflows/sarek.nf +++ b/workflows/sarek.nf @@ -789,7 +789,8 @@ workflow SAREK { pon, pon_tbi, chr_files, - mappability + mappability, + [] ) // PAIR VARIANT CALLING @@ -810,7 +811,8 @@ workflow SAREK { pon, pon_tbi, chr_files, - mappability + mappability, + [] ) // Gather vcf files for annotation and QC From 1fbb4efb22be5bab478ece5d901d4722a42d5792 Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 20 Jun 2022 08:18:39 +0200 Subject: [PATCH 2/8] Updated Changelog --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index bdc8e1fc8..7fdad090e 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -30,7 +30,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#557](https://github.com/nf-core/sarek/pull/557) - Add `Haplotypecaller` single sample mode together with `CNNScoreVariants` and `FilterVariantTranches` - [#576](https://github.com/nf-core/sarek/pull/576) - Add modules and subworkflows for `cnvkit` germline mode - [#582](https://github.com/nf-core/sarek/pull/582) - Added option `--vep_out_format` for setting the format of the output-file from VEP to `json`, `tab` or `vcf` (default) - +- [#597](https://github.com/nf-core/sarek/pull/597) - Added tiddit for pair and tumor variant calling ### Changed - [#383](https://github.com/nf-core/sarek/pull/383), [#528](https://github.com/nf-core/sarek/pull/528) - Update `CHANGELOG` From 851cf9bfbc72c0f0d2647f6bb784236ea94f47f6 Mon Sep 17 00:00:00 2001 From: WackerO Date: Mon, 20 Jun 2022 08:54:43 +0200 Subject: [PATCH 3/8] Ran prettier --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 7fdad090e..8cc362e03 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -31,6 +31,7 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0 - [#576](https://github.com/nf-core/sarek/pull/576) - Add modules and subworkflows for `cnvkit` germline mode - [#582](https://github.com/nf-core/sarek/pull/582) - Added option `--vep_out_format` for setting the format of the output-file from VEP to `json`, `tab` or `vcf` (default) - [#597](https://github.com/nf-core/sarek/pull/597) - Added tiddit for pair and tumor variant calling + ### Changed - [#383](https://github.com/nf-core/sarek/pull/383), [#528](https://github.com/nf-core/sarek/pull/528) - Update `CHANGELOG` From af4cbf3ac8db3ff79dc01a2b9d7673558b621629 Mon Sep 17 00:00:00 2001 From: WackerO <43847497+WackerO@users.noreply.github.com> Date: Mon, 20 Jun 2022 09:46:58 +0200 Subject: [PATCH 4/8] Update subworkflows/local/pair_variant_calling.nf Co-authored-by: Maxime U. Garcia --- subworkflows/local/pair_variant_calling.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/subworkflows/local/pair_variant_calling.nf b/subworkflows/local/pair_variant_calling.nf index c2d25df09..c45e2df2c 100644 --- a/subworkflows/local/pair_variant_calling.nf +++ b/subworkflows/local/pair_variant_calling.nf @@ -43,7 +43,7 @@ workflow PAIR_VARIANT_CALLING { strelka_vcf = Channel.empty() msisensorpro_output = Channel.empty() mutect2_vcf = Channel.empty() - tiddit_vcf = Channel.empty() + tiddit_vcf = Channel.empty() // Remap channel with intervals cram_pair_intervals = cram_pair.combine(intervals) From c71b81b69b97293efddb6ff8dfedfeb34f5a0b86 Mon Sep 17 00:00:00 2001 From: WackerO Date: Wed, 22 Jun 2022 10:06:29 +0200 Subject: [PATCH 5/8] Removed pairVC tiddit; will look at this in another PR --- subworkflows/local/pair_variant_calling.nf | 16 ++-------------- subworkflows/local/tumor_variant_calling.nf | 3 --- tests/test_tools.yml | 14 -------------- workflows/sarek.nf | 3 +-- 4 files changed, 3 insertions(+), 33 deletions(-) diff --git a/subworkflows/local/pair_variant_calling.nf b/subworkflows/local/pair_variant_calling.nf index c45e2df2c..49d0723d2 100644 --- a/subworkflows/local/pair_variant_calling.nf +++ b/subworkflows/local/pair_variant_calling.nf @@ -10,7 +10,6 @@ include { RUN_STRELKA_SOMATIC } from '../nf-core/variantca include { RUN_CNVKIT_SOMATIC } from '../nf-core/variantcalling/cnvkit/somatic/main.nf' include { RUN_MPILEUP as RUN_MPILEUP_NORMAL } from '../nf-core/variantcalling/mpileup/main' include { RUN_MPILEUP as RUN_MPILEUP_TUMOR } from '../nf-core/variantcalling/mpileup/main' -include { RUN_TIDDIT } from '../nf-core/variantcalling/tiddit/main.nf' workflow PAIR_VARIANT_CALLING { take: @@ -31,7 +30,6 @@ workflow PAIR_VARIANT_CALLING { panel_of_normals_tbi // channel: [optional] panel_of_normals_tbi chr_files mappability - bwa // channel: [optional] bwa main: @@ -43,7 +41,6 @@ workflow PAIR_VARIANT_CALLING { strelka_vcf = Channel.empty() msisensorpro_output = Channel.empty() mutect2_vcf = Channel.empty() - tiddit_vcf = Channel.empty() // Remap channel with intervals cram_pair_intervals = cram_pair.combine(intervals) @@ -191,15 +188,8 @@ workflow PAIR_VARIANT_CALLING { ch_versions = ch_versions.mix(GATK_TUMOR_NORMAL_SOMATIC_VARIANT_CALLING.out.versions) } - //TIDDIT - if (tools.contains('tiddit')){ - RUN_TIDDIT(cram_pair, - fasta, - bwa) - - tiddit_vcf = RUN_TIDDIT.out.tiddit_vcf - ch_versions = ch_versions.mix(RUN_TIDDIT.out.versions) - } + // if (tools.contains('tiddit')) { + // } emit: freebayes_vcf @@ -207,7 +197,5 @@ workflow PAIR_VARIANT_CALLING { msisensorpro_output mutect2_vcf strelka_vcf - tiddit_vcf - versions = ch_versions } diff --git a/subworkflows/local/tumor_variant_calling.nf b/subworkflows/local/tumor_variant_calling.nf index 210e93e3e..a2dd68cd2 100644 --- a/subworkflows/local/tumor_variant_calling.nf +++ b/subworkflows/local/tumor_variant_calling.nf @@ -166,9 +166,6 @@ workflow TUMOR_ONLY_VARIANT_CALLING { } - // if (tools.contains('tiddit')){ - // } - emit: freebayes_vcf manta_vcf diff --git a/tests/test_tools.yml b/tests/test_tools.yml index f593f69b5..bebe16bb3 100644 --- a/tests/test_tools.yml +++ b/tests/test_tools.yml @@ -92,20 +92,6 @@ - path: results/variant_calling/sample2/tiddit/sample2.vcf.gz - path: results/variant_calling/sample2/tiddit/sample2.vcf.gz.tbi -- name: Run variant calling on somatic sample with tiddit - command: nextflow run main.nf -profile test,tools_somatic,docker --tools tiddit -c ./tests/nextflow.config - tags: - - tiddit - - somatic - - variant_calling - files: - - path: results/variant_calling/sample4_vs_sample3/tiddit/sample4_vs_sample3.ploidies.tab - - path: results/variant_calling/sample4_vs_sample3/tiddit/sample4_vs_sample3.vcf.gz - - path: results/variant_calling/sample4_vs_sample3/tiddit/sample4_vs_sample3.vcf.gz.tbi - - path: results/variant_calling/sample3/tiddit/sample3.ploidies.tab - - path: results/variant_calling/sample3/tiddit/sample3.vcf.gz - - path: results/variant_calling/sample3/tiddit/sample3.vcf.gz.tbi - - name: Run variant calling on somatic samples with controlfreec command: nextflow run main.nf -profile test,tools_somatic,docker --tools controlfreec -c ./tests/nextflow.config tags: diff --git a/workflows/sarek.nf b/workflows/sarek.nf index 256e72239..79df15794 100644 --- a/workflows/sarek.nf +++ b/workflows/sarek.nf @@ -811,8 +811,7 @@ workflow SAREK { pon, pon_tbi, chr_files, - mappability, - [] + mappability ) // Gather vcf files for annotation and QC From 94c5f347bdedb233f1723182a6f69578a1dd5a48 Mon Sep 17 00:00:00 2001 From: WackerO Date: Wed, 22 Jun 2022 11:39:49 +0200 Subject: [PATCH 6/8] Updated subway map --- docs/images/sarek_subway.png | Bin 138462 -> 140608 bytes docs/images/sarek_subway.svg | 150 ++++++++++++++++++++++------------- 2 files changed, 95 insertions(+), 55 deletions(-) diff --git a/docs/images/sarek_subway.png 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inkscape:window-x="0" + inkscape:zoom="1.4127647" + inkscape:cx="577.23696" + inkscape:cy="353.56206" + inkscape:window-width="1600" + inkscape:window-height="947" + inkscape:window-x="1024" inkscape:window-y="25" inkscape:window-maximized="1" inkscape:current-layer="layer4" @@ -1024,7 +1024,7 @@ + transform="translate(-109.26933,-3.7167545)"> + transform="translate(-109.06183,122.0378)"> + style="opacity:1;fill:none;fill-opacity:1;stroke:#000000;stroke-width:4.00201;stroke-linecap:butt;stroke-linejoin:bevel;stroke-miterlimit:4;stroke-dasharray:none;stroke-dashoffset:0;stroke-opacity:1;paint-order:normal" /> tiddit + tiddit ascat msisensorpro controlfreec cnvkit + @@ -2674,35 +2696,35 @@ @@ -2737,7 +2759,7 @@ @@ -2785,7 +2807,7 @@ style="color:#000000;font-style:normal;font-variant:normal;font-weight:normal;font-stretch:normal;font-size:medium;line-height:normal;font-family:sans-serif;font-variant-ligatures:normal;font-variant-position:normal;font-variant-caps:normal;font-variant-numeric:normal;font-variant-alternates:normal;font-feature-settings:normal;text-indent:0;text-align:start;text-decoration:none;text-decoration-line:none;text-decoration-style:solid;text-decoration-color:#000000;letter-spacing:normal;word-spacing:normal;text-transform:none;writing-mode:lr-tb;direction:ltr;text-orientation:mixed;dominant-baseline:auto;baseline-shift:baseline;text-anchor:start;white-space:normal;shape-padding:0;clip-rule:nonzero;display:inline;overflow:visible;visibility:visible;opacity:1;isolation:auto;mix-blend-mode:normal;color-interpolation:sRGB;color-interpolation-filters:linearRGB;solid-color:#000000;solid-opacity:1;vector-effect:none;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;stroke-width:4.00004;stroke-linecap:butt;stroke-linejoin:miter;stroke-miterlimit:4;stroke-dasharray:none;stroke-dashoffset:0;stroke-opacity:1;color-rendering:auto;image-rendering:auto;shape-rendering:auto;text-rendering:auto;enable-background:accumulate" /> + + + + Date: Thu, 23 Jun 2022 08:53:17 +0200 Subject: [PATCH 7/8] Added tiddit vcf to annotation --- workflows/sarek.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/workflows/sarek.nf b/workflows/sarek.nf index 1e19cf8e9..e371c82f2 100644 --- a/workflows/sarek.nf +++ b/workflows/sarek.nf @@ -856,6 +856,7 @@ workflow SAREK { vcf_to_annotate = vcf_to_annotate.mix(TUMOR_ONLY_VARIANT_CALLING.out.mutect2_vcf) vcf_to_annotate = vcf_to_annotate.mix(TUMOR_ONLY_VARIANT_CALLING.out.manta_vcf) vcf_to_annotate = vcf_to_annotate.mix(TUMOR_ONLY_VARIANT_CALLING.out.strelka_vcf) + vcf_to_annotate = vcf_to_annotate.mix(TUMOR_ONLY_VARIANT_CALLING.out.tiddit_vcf) vcf_to_annotate = vcf_to_annotate.mix(PAIR_VARIANT_CALLING.out.mutect2_vcf) vcf_to_annotate = vcf_to_annotate.mix(PAIR_VARIANT_CALLING.out.manta_vcf) vcf_to_annotate = vcf_to_annotate.mix(PAIR_VARIANT_CALLING.out.strelka_vcf) From 656af027eb470b4ebe6ebaf75dd27994ee605103 Mon Sep 17 00:00:00 2001 From: WackerO Date: Thu, 23 Jun 2022 13:23:55 +0200 Subject: [PATCH 8/8] Sorted input files --- subworkflows/local/germline_variant_calling.nf | 4 ++-- subworkflows/local/tumor_variant_calling.nf | 10 +++++----- workflows/sarek.nf | 18 +++++++++--------- 3 files changed, 16 insertions(+), 16 deletions(-) diff --git a/subworkflows/local/germline_variant_calling.nf b/subworkflows/local/germline_variant_calling.nf index ee0d45c72..9096f68b8 100644 --- a/subworkflows/local/germline_variant_calling.nf +++ b/subworkflows/local/germline_variant_calling.nf @@ -18,14 +18,14 @@ workflow GERMLINE_VARIANT_CALLING { bwa // channel: [mandatory] bwa dbsnp // channel: [mandatory] dbsnp dbsnp_tbi // channel: [mandatory] dbsnp_tbi - known_sites - known_sites_tbi dict // channel: [mandatory] dict fasta // channel: [mandatory] fasta fasta_fai // channel: [mandatory] fasta_fai intervals // channel: [mandatory] intervals/target regions intervals_bed_gz_tbi // channel: [mandatory] intervals/target regions index zipped and indexed intervals_bed_combined // channel: [mandatory] intervals/target regions in one file unzipped + known_sites + known_sites_tbi // joint_germline // val: true/false on whether to run joint_germline calling, only works in combination with haplotypecaller at the moment main: diff --git a/subworkflows/local/tumor_variant_calling.nf b/subworkflows/local/tumor_variant_calling.nf index a2dd68cd2..f9c8b44f0 100644 --- a/subworkflows/local/tumor_variant_calling.nf +++ b/subworkflows/local/tumor_variant_calling.nf @@ -16,21 +16,21 @@ workflow TUMOR_ONLY_VARIANT_CALLING { take: tools // Mandatory, list of tools to apply cram_recalibrated // channel: [mandatory] cram + bwa // channel: [optional] bwa + chr_files dbsnp // channel: [mandatory] dbsnp dbsnp_tbi // channel: [mandatory] dbsnp_tbi dict // channel: [mandatory] dict fasta // channel: [mandatory] fasta fasta_fai // channel: [mandatory] fasta_fai + germline_resource // channel: [optional] germline_resource + germline_resource_tbi // channel: [optional] germline_resource_tbi intervals // channel: [mandatory] intervals/target regions intervals_bed_gz_tbi // channel: [mandatory] intervals/target regions index zipped and indexed intervals_bed_combined // channel: [mandatory] intervals/target regions in one file unzipped - germline_resource // channel: [optional] germline_resource - germline_resource_tbi // channel: [optional] germline_resource_tbi + mappability panel_of_normals // channel: [optional] panel_of_normals panel_of_normals_tbi // channel: [optional] panel_of_normals_tbi - chr_files - mappability - bwa // channel: [optional] bwa main: diff --git a/workflows/sarek.nf b/workflows/sarek.nf index e371c82f2..28750c994 100644 --- a/workflows/sarek.nf +++ b/workflows/sarek.nf @@ -792,35 +792,35 @@ workflow SAREK { [], dbsnp, dbsnp_tbi, - known_sites, - known_sites_tbi, dict, fasta, fasta_fai, intervals, intervals_bed_gz_tbi, - intervals_bed_combined) + intervals_bed_combined, + known_sites, + known_sites_tbi) // params.joint_germline) // TUMOR ONLY VARIANT CALLING TUMOR_ONLY_VARIANT_CALLING( params.tools, cram_variant_calling_tumor_only, + [], + chr_files, dbsnp, dbsnp_tbi, dict, fasta, fasta_fai, + germline_resource, + germline_resource_tbi, intervals, intervals_bed_gz_tbi, intervals_bed_combined, - germline_resource, - germline_resource_tbi, - pon, - pon_tbi, - chr_files, mappability, - [] + pon, + pon_tbi ) // PAIR VARIANT CALLING