Feat indelrealign

conf/base.config

@@ -46,7 +46,9 @@ process {
         time   = { check_max( 16.h   * task.attempt, 'time'    ) }
     }
     withLabel:process_long {
-        time   = { check_max( 20.h  * task.attempt, 'time'    ) }
+        cpus   = { check_max( 4     * task.attempt, 'cpus'    ) }
+        memory = { check_max( 16.GB * task.attempt, 'memory'  ) }
+        time   = { check_max( 72.h  * task.attempt, 'time'    ) }
     }
     withLabel:process_high_memory {
         memory = { check_max( 200.GB * task.attempt, 'memory' ) }

conf/genomes.config

@@ -7,12 +7,12 @@ params {
             genomedict= "/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/genome/Homo_sapiens_assembly38.dict"
             wgsregion = "/data/CCBR_Pipeliner/Pipelines/LOGAN/resources/hg38/resources_broad_hg38_v0_wgs_calling_regions.hg38.interval_list"
             intervals= "${projectDir}/assets/hg38_v0_wgs_calling_regions.hg38.bed"
-            //millsindel = "/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/Mills_and_1000G_gold_standard.indels.hg38.vcf.gz" //Mills_and_1000G_gold_standard.indels.hg38.vcf.gz"
-            //shapeitindel =  "/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/ALL.wgs.1000G_phase3.GRCh38.ncbi_remapper.20150424.shapeit2_indels.vcf.gz" //ALL.wgs.1000G_phase3.GRCh38.ncbi_remapper.20150424.shapeit2_indels.vcf.gz" //file(params.gold_indels2) //
-            KNOWNINDELS= '/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/Mills_and_1000G_gold_standard.indels.hg38.vcf.gz'
+            millsindel = "/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/Mills_and_1000G_gold_standard.indels.hg38.vcf.gz" //Mills_and_1000G_gold_standard.indels.hg38.vcf.gz"
+            shapeitindel =  "/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/ALL.wgs.1000G_phase3.GRCh38.ncbi_remapper.20150424.shapeit2_indels.vcf.gz" //ALL.wgs.1000G_phase3.GRCh38.ncbi_remapper.20150424.shapeit2_indels.vcf.gz" //file(params.gold_indels2) //
+            INDELREF = "/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/ALL.wgs.1000G_phase3.GRCh38.ncbi_remapper.20150424.shapeit2_indels.vcf.gz" //ALL.wgs.1000G_phase3.GRCh38.ncbi_remapper.20150424.shapeit2_indels.vcf.gz" 
+            KNOWNINDELS = "-known /data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/Mills_and_1000G_gold_standard.indels.hg38.vcf.gz -known /data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/ALL.wgs.1000G_phase3.GRCh38.ncbi_remapper.20150424.shapeit2_indels.vcf.gz"
             KNOWNRECAL = '--known-sites /data/CCBR_Pipeliner/Pipelines/XAVIER/resources/hg38/GATK_resource_bundle/dbsnp_138.hg38.vcf.gz --known-sites /data/CCBR_Pipeliner/Pipelines/XAVIER/resources/hg38/GATK_resource_bundle/Mills_and_1000G_gold_standard.indels.hg38.vcf.gz --known-sites /data/CCBR_Pipeliner/Pipelines/XAVIER/resources/hg38/GATK_resource_bundle/ALL.wgs.1000G_phase3.GRCh38.ncbi_remapper.20150424.shapeit2_indels.vcf.gz'
             dbsnp = "/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/dbsnp_138.hg38.vcf.gz"
-            dbsnp_indel = "/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/Mills_and_1000G_gold_standard.indels.hg38.vcf.gz"
             gnomad = '--germline-resource /data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GNOMAD/somatic-hg38-af-only-gnomad.hg38.vcf.gz' // /data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GNOMAD/somatic-hg38-af-only-gnomad.hg38.vcf.gz
             pon = "/data/CCBR_Pipeliner/Pipelines/LOGAN/resources/hg38/PON/updatedpon.vcf.gz"    //pon="/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/PON/hg38.noCOSMIC_ClinVar.pon.vcf.gz" //file{params.pon}
             kgp = "/data/CCBR_Pipeliner/CCBR_Pipeliner_Legacy/Exome-seek/hg38/GATK_resource_bundle/1000G_phase1.snps.high_confidence.hg38.vcf.gz"
@@ -25,6 +25,7 @@ params {
             vepcache = "/fdb/VEP/102/cache"
             vepspecies = "homo_sapiens"
             vepbuild = "GRCh38"
+            annotsvgenome = "GRCh38"
             octopus_sforest= "--somatic-forest /data/CCBR_Pipeliner/Pipelines/LOGAN/resources/hg38/octopus/somatic.v0.7.4.forest"
             octopus_gforest= "--forest /data/CCBR_Pipeliner/Pipelines/LOGAN/resources/hg38/octopus/germline.v0.7.4.forest"
             SEQUENZAGC = "/data/CCBR_Pipeliner/Pipelines/XAVIER/resources/hg38/SEQUENZA/hg38_gc50Base.txt.gz"
@@ -37,7 +38,8 @@ params {
             bwagenome= "/data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/genome/bwaindex/genome.fa"
             genomedict= "/data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/genome/bwamem2index/genome.dict"
             intervals="/data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/genome/bwamem2index/mm10_wgsregions.bed"
-            KNOWNINDELS = "/data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/dbsnp/mm10_known_indels.vcf.gz"
+            KNOWNINDELS = "-known /data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/dbsnp/mm10_known_indels.vcf.gz"
+            INDELREF = "/data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/dbsnp/mm10_known_indels.vcf.gz"
             KNOWNRECAL = "-known-sites /data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/dbsnp/mm10_known_indels.vcf.gz -known-sites /data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/dbsnp/mm10_known_snps.vcf.gz"
             dbsnp = "/data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/dbsnp/mm10_allstrains_dbSNP142.vcf.gz"
             pon = "/data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/dbsnp/mm10_dbSNP_allStrains_compSet_noIND.vcf.gz"
@@ -53,6 +55,7 @@ params {
             vepcache = "/fdb/VEP/102/cache"
             vepspecies = "mus_musculus"
             vepbuild= "GRCm38"
+            annotsvgenome = "mm10"
             octopus_sforest = ""
             octopus_gforest = ""
 	 	    SEQUENZAGC =  '/data/CCBR_Pipeliner/Pipelines/XAVIER/resources/mm10/SEQUENZA/mm10.gc50Base.wig.gz'

main.nf

@@ -25,8 +25,6 @@ include {INPUT_TONLY; INPUT_TONLY_BAM;
     VC_TONLY; SV_TONLY; CNVhuman_tonly; CNVmouse_tonly; QC_TONLY } from "./subworkflows/local/workflows_tonly.nf"
 
 
-
-//SUB WORKFLOWS to SPLIT
 workflow.onComplete {
     if (!workflow.stubRun && !workflow.commandLine.contains('-preview')) {
         def message = Utils.spooker(workflow)

modules/local/copynumber.nf

@@ -21,7 +21,6 @@ ENSEMBLCACHE='/data/SCLC-BRAINMETS/cn/common/ensembl_data'
 DRIVERS='/data/SCLC-BRAINMETS/cn/common/DriverGenePanel.38.tsv'
 HOTSPOTS='/data/SCLC-BRAINMETS/cn/variants/KnownHotspots.somatic.38.vcf.gz'
 
-//DBSNP_INDEL=file(params.genomes[params.genome].KNOWNINDELS)
 //ascatR=
 
 
@@ -273,7 +272,7 @@ process amber_tonly {
 
     """
 
-    java -Xmx32G -cp amber.jar com.hartwig.hmftools.amber.AmberApplication \
+    java -Xmx32G -cp /opt2/hmftools/amber.jar com.hartwig.hmftools.amber.AmberApplication \
     -tumor ${tumorname} -tumor_bam ${tumor} \
     -output_dir ${tumorname}_amber \
     -threads $task.cpus \
@@ -310,7 +309,7 @@ process amber_tn {
 
     """
 
-    java -Xmx32G -cp amber.jar com.hartwig.hmftools.amber.AmberApplication \
+    java -Xmx32G -cp /opt2/hmftools/amber.jar com.hartwig.hmftools.amber.AmberApplication \
     -tumor ${tumorname} -tumor_bam ${tumor} \
     -reference ${normalname} -reference_bam ${normal} \
     -output_dir ${tumorname}_vs_${normalname}_amber \
@@ -346,7 +345,7 @@ process cobalt_tonly {
 
     """
 
-    java -jar -Xmx8G cobalt.jar \
+    java -jar -Xmx8G /opt2/hmftools/cobalt.jar \
     -tumor ${tumorname} -tumor_bam ${tumor} \
     -output_dir ${tumorname}_cobalt \
     -threads $task.cpus \
@@ -382,7 +381,7 @@ process cobalt_tn {
 
     """
 
-    java -jar -Xmx8G cobalt.jar \
+    java -jar -Xmx8G /opt2/hmftools/cobalt.jar \
     -tumor ${tumorname} -tumor_bam ${tumorname} \
     -reference ${normalname} -reference_bam ${normal} \
     -output_dir ${tumorname}_vs_${normalname}_cobalt \
@@ -418,7 +417,7 @@ process purple {
     script:
 
     """
-    java -jar purple.jar \
+    java -jar /opt2/hmftools/purple.jar \
     -tumor ${tumorname} \
     -amber ${amberin} \
     -cobalt ${cobaltin} \

modules/local/structural_variant.nf

@@ -1,11 +1,12 @@
 GENOMEREF=file(params.genomes[params.genome].genome)
-GENOME=params.genome
+ANNOTSVGENOME=file(params.genomes[params.genome].annotsvgenome)
 BWAGENOME=file(params.genomes[params.genome].bwagenome)
-DBSNP_INDEL=file(params.genomes[params.genome].KNOWNINDELS)
+INDELREF=file(params.genomes[params.genome].INDELREF)
 
 
 
 process svaba_somatic {
+    container = "${params.containers.logan}"
     label 'process_highcpu'
 
     input:
@@ -30,7 +31,7 @@ process svaba_somatic {
 
     script:
     """
-    svaba run -t ${tumor} -n ${normal} -p $task.cpus -D $DBSNP_INDEL -a ${tumor.simpleName} -G $BWAGENOME
+    svaba run -t ${tumor} -n ${normal} -p $task.cpus -D $INDELREF -a ${tumor.simpleName} -G $BWAGENOME
     """
 
     stub:
@@ -56,7 +57,7 @@ process svaba_somatic {
 
 
 process manta_somatic {
-
+    container = "${params.containers.logan}"
     label 'process_highcpu'
 
     input:
@@ -102,7 +103,6 @@ process manta_somatic {
 process annotsv_tn {
      //AnnotSV for Manta/Svaba works with either vcf.gz or .vcf files
      //Requires bedtools,bcftools
-
     module = ['annotsv/3.3.1']
 
     input:
@@ -119,7 +119,7 @@ process annotsv_tn {
     mkdir ${sv}
 
     AnnotSV -SVinputFile ${somaticvcf} \
-    -genomeBuild $GENOME \
+    -genomeBuild $ANNOTSVGENOME \
     -SVinputInfo 1 -outputFile ${tumorname} \
     -outputDir ${sv}
 
@@ -136,6 +136,7 @@ process annotsv_tn {
 
 
 process manta_tonly {
+    container = "${params.containers.logan}"
     label 'process_highcpu'
 
     input:
@@ -178,6 +179,7 @@ process manta_tonly {
 
 
 process svaba_tonly {
+    container = "${params.containers.logan}"
     label 'process_highcpu'
 
     input:
@@ -198,7 +200,7 @@ process svaba_tonly {
 
     script:
     """
-    svaba run -t ${tumor} -p $task.cpus -D $DBSNP_INDEL -a ${tumor.simpleName} -G $BWAGENOME
+    svaba run -t ${tumor} -p $task.cpus -D $INDELREF -a ${tumor.simpleName} -G $BWAGENOME
     """
 
     stub:
@@ -230,7 +232,7 @@ process gunzip {
 
     script:
     """
-    gunzip ${vcf} > ${tumorname}.tumorSV.vcf
+    gunzip -f ${vcf} > ${tumorname}.tumorSV.vcf
     """
 
     stub:
@@ -291,7 +293,7 @@ process annotsv_tonly {
     mkdir ${sv}
 
     AnnotSV -SVinputFile ${somaticvcf} \
-    -genomeBuild $GENOME \
+    -genomeBuild $ANNOTSVGENOME \
     -SVinputInfo 1 -outputFile ${tumorname} \
     -outputDir ${sv}
 

modules/local/trim_align.nf

@@ -1,6 +1,6 @@
-GENOMEREF=file(params.genomes[params.genome].genome)
+GENOMEREF = file(params.genomes[params.genome].genome)
 KNOWNRECAL = params.genomes[params.genome].KNOWNRECAL
-
+KNOWNINDELS = params.genomes[params.genome].KNOWNINDELS
 
 process fastp {
     container = "${params.containers.logan}"
@@ -77,6 +77,70 @@ process bwamem2 {
 
 
 
+process indelrealign {
+    container "${params.containers.logan}"
+    label 'process_long'
+
+    input:
+    tuple val(samplename), path("${samplename}.bam"), path("${samplename}.bai")
+
+    output:
+    tuple val(samplename), path("${samplename}.ir.bam"), path("${samplename}.ir.bai")
+
+    script:
+
+    """
+    /usr/lib/jvm/java-8-openjdk-amd64/bin/java -Xmx16g -jar \$GATK_JAR -T RealignerTargetCreator \
+        -I ${samplename}.bam \
+        -R ${GENOMEREF} \
+        -o ${samplename}.intervals \
+        -nt $task.cpus \
+        ${KNOWNINDELS}
+
+    /usr/lib/jvm/java-8-openjdk-amd64/bin/java -Xmx16g -jar \$GATK_JAR -T IndelRealigner \
+        -R ${GENOMEREF} \
+        -I ${samplename}.bam \
+        ${KNOWNINDELS} \
+        -targetIntervals ${samplename}.intervals \
+        -o ${samplename}.ir.bam
+    """
+
+    stub:
+    """
+    touch ${samplename}.ir.bam ${samplename}.ir.bai
+    """
+
+}
+
+
+process bqsr_ir {
+    /*
+    Base quality recalibration for all samples
+    */
+    container = "${params.containers.logan}"
+    label 'process_low'
+    input:
+        tuple val(samplename), path("${samplename}.ir.bam"), path("${samplename}.ir.bai"), path(bed)
+
+    output:
+        tuple val(samplename), path("${samplename}_${bed.simpleName}.recal_data.grp")
+
+    script:
+    """
+    gatk --java-options '-Xmx16g' BaseRecalibrator \
+    --input ${samplename}.ir.bam \
+    --reference ${GENOMEREF} \
+    ${KNOWNRECAL} \
+    --output ${samplename}_${bed.simpleName}.recal_data.grp \
+    --intervals ${bed}
+    """
+
+    stub:
+    """
+    touch ${samplename}_${bed.simpleName}.recal_data.grp
+    """
+}
+
 process bqsr {
     /*
     Base quality recalibration for all samples
@@ -103,7 +167,6 @@ process bqsr {
     """
     touch ${samplename}_${bed.simpleName}.recal_data.grp
     """
-
 }
 
 process gatherbqsr {
@@ -131,16 +194,15 @@ process gatherbqsr {
     """
 }
 
-
 process applybqsr {
     /*
     Base quality recalibration for all samples to
     */
     container = "${params.containers.logan}"
-    label 'process_low'
+    label 'process_long'
 
     input:
-        tuple val(samplename), path("${samplename}.bam"), path("${samplename}.bai"), path("${samplename}.recal_data.grp")
+        tuple val(samplename), path(bam), path(bai), path("${samplename}.recal_data.grp")
 
     output:
         tuple val(samplename), path("${samplename}.bqsr.bam"),  path("${samplename}.bqsr.bai")
@@ -150,7 +212,7 @@ process applybqsr {
     """
     gatk --java-options '-Xmx32g' ApplyBQSR \
         --reference ${GENOMEREF} \
-        --input ${samplename}.bam \
+        --input ${bam} \
         --bqsr-recal-file ${samplename}.recal_data.grp \
         --output ${samplename}.bqsr.bam \
         --use-jdk-inflater \
@@ -166,7 +228,6 @@ process applybqsr {
 }
 
 
-
 process samtoolsindex {
     container = "${params.containers.logan}"
     label 'process_medium'
@@ -198,48 +259,21 @@ process bamtocram_tonly {
         tuple val(tumorname), path(tumor), path(tumorbai)
 
     output:
-        path("${sample}.cram")
-
-    script:
-    """
-        samtools view -@ $task.cpus -C -T $GENOMEREF -o ${sample}.cram {$tumor}.bam
-    """
-}
-
+        path("${tumorname}.cram"), path("${tumorname}.cram.crai")
 
-/*
-process indelrealign {
-    input:
-    tuple val(samplename), path("${samplename}.bam"), path("${samplename}.bai")
-
-    output:
-    tuple val(samplename), path("${samplename}.ir.bam")
 
     script:
-
     """
-    /usr/bin/java -Xmx32g -jar \${GATK_JAR} -T RealignerTargetCreator \
-        -I ${samplename}.bam \
-        -R ${GENOMEREF} \
-        -o ${samplename}.intervals \
-        -nt 16 \
-        -known ${MILLSINDEL} -known ${SHAPEITINDEL}
-
-    /usr/bin/java -Xmx32g -jar \${GATK_JAR} -T IndelRealigner \
-        -R ${GENOMEREF} \
-        -I ${samplename}.bam \
-        -known ${MILLSINDEL} -known ${SHAPEITINDEL} \
-        --use_jdk_inflater \
-        --use_jdk_deflater \
-        -targetIntervals ${samplename}.intervals \
-        -o  ${samplename}.ir.bam
+        samtools view -@ $task.cpus -C -T $GENOMEREF -o ${sample}.cram $tumor
+        samtools index ${tumorname}.cram -@ $task.cpus
     """
-
-
+
     stub:
     """
-    touch ${samplename}.ir.bam
+    touch ${tumorname}.cram ${tumorname}.cram.crai
     """
-
 }
-*/
+
+
+
+