963 lines
30 KiB
Plaintext
963 lines
30 KiB
Plaintext
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workflow pancancer_controlsample{
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String normal
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String tumor
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String cancer
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String project = "650gene"
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String bed = "/dataseq/jmdna/database/bed/650.bed"
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String bed_plus = "/dataseq/jmdna/database/bed/650plus50bp.bed"
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String name
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String outputDir
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String need_TMB_test = "yes"
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String need_MSI_test = "yes"
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String need_neoantigen_test = "yes"
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String inputDir
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String ref = "/dataseq/jmdna/database/genome/hg19/hg19.fa"
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String codes_dir = "/dataseq/jmdna/codes/pancancer_controlsample"
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String accessBed = "/dataseq/jmdna/software/cnvkit-0.9.7/data/access-5k-mappable.hg19.bed"
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String annotateGene = "/dataseq/jmdna/software/cnvkit-0.9.7/data/refFlat.txt"
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String gc_wiggle = "/dataseq/jmdna/codes/pancancer_controlsample/hg19.gc200Base.txt.gz"
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Boolean get_sampleinfo_local = false
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#创建目录
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call create_dir{
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input:
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outputDir=outputDir,
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need_neoantigen_test=need_neoantigen_test,
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need_MSI_test=need_MSI_test
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}
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scatter (name in [normal,tumor]){
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call qc{
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input:
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name=name,
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inputDir=inputDir,
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outputDir=outputDir
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}
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call alignment_bwa{
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input:
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name=name,
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outputDir=outputDir,
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ref=ref,
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read1=qc.outputFile[0],
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read2=qc.outputFile[1]
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}
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call markduplicates{
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input:
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name=name,
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outputDir=outputDir,
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ref=ref,
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sortedBam=alignment_bwa.sorted
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}
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call generater_mpileup{
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input:
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name=name,
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ref=ref,
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bed=bed_plus,
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outputDir=outputDir,
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rmdupBam=markduplicates.rmdup
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}
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}
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call mutation_calling{
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input:
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codes_dir=codes_dir,
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tumor=tumor,
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outputDir=outputDir,
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pileup=generater_mpileup.pileup
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}
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call annovar{
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input:
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tumor=tumor,
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normal=normal,
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outputDir=outputDir,
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ref=ref,
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vcf=mutation_calling.vcf
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}
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call dealwithsnvindel{
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input:
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codes_dir=codes_dir,
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project=project,
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outputDir=outputDir,
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tumor=tumor,
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cancer=cancer,
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anno=annovar.anno
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}
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call fusion{
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input:
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ref=ref,
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codes_dir=codes_dir,
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project=project,
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tumor=tumor,
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outputDir=outputDir,
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cancer=cancer,
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rmdupBam=markduplicates.rmdup
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}
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if (need_TMB_test=="yes"){
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call TMB{
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input:
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codes_dir=codes_dir,
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tumor=tumor,
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outputDir=outputDir,
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anno=annovar.anno,
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concordance=conpair.concordance,
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snv_result=dealwithsnvindel.snv,
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cnv_result=cnvkit.cnv,
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fusion_result=fusion.fusion
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}
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}
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if (need_MSI_test=="yes"){
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call MSI{
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input:
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bed=bed,
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tumor=tumor,
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normal=normal,
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outputDir=outputDir,
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rmdupBam=markduplicates.rmdup
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}
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}
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call MMR{
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input:
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codes_dir=codes_dir,
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tumor=tumor,
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outputDir=outputDir,
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snv=dealwithsnvindel.snv
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}
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call HRR{
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input:
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codes_dir=codes_dir,
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tumor=tumor,
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outputDir=outputDir,
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snv=dealwithsnvindel.snv
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}
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call chemoTherapy{
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input:
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codes_dir=codes_dir,
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normal=normal,
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tumor=tumor,
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outputDir=outputDir,
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ref=ref,
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project=project,
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rmdupBam=markduplicates.rmdup
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}
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call hereditary{
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input:
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codes_dir=codes_dir,
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tumor=tumor,
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outputDir=outputDir,
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project=project,
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snv=dealwithsnvindel.snv
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}
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call conpair{
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input:
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name=name,
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codes_dir=codes_dir,
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tumor=tumor,
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normal=normal,
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ref=ref,
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outputDir=outputDir,
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rmdupBam=markduplicates.rmdup
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}
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call cnvkit{
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input:
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name=name,
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normal=normal,
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tumor=tumor,
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ref=ref,
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bed=bed,
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cancer=cancer,
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project=project,
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codes_dir=codes_dir,
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outputDir=outputDir,
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rmdupBam=markduplicates.rmdup,
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accessBed=accessBed,
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annotateGene=annotateGene,
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purity=tumor_content.purity
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}
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call tumor_content{
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input:
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name=name,
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ref=ref,
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tumor=tumor,
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normal=normal,
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outputDir=outputDir,
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gc_wiggle=gc_wiggle,
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codes_dir=codes_dir,
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pileup=generater_mpileup.pileup
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}
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if (need_neoantigen_test=="yes"){
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call HLA{
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input:
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inputDir=inputDir,
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outputDir=outputDir,
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normal=normal,
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newdir=create_dir.newdir
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}
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call neoantigen{
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input:
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codes_dir=codes_dir,
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outputDir=outputDir,
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name=name,
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normal=normal,
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tumor=tumor,
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hla=HLA.hla,
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vcf=mutation_calling.vcf
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}
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}
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scatter (name in [normal,tumor]){
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call qc_2{
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input:
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ref=ref,
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bed=bed,
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codes_dir=codes_dir,
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name=name,
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tumor=tumor,
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normal=normal,
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outputDir=outputDir,
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anno=annovar.anno,
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get_sampleinfo_local=get_sampleinfo_local
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}
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}
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call auto_report{
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input:
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cancer=cancer,
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codes_dir=codes_dir,
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outputDir=outputDir,
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name=name,
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normal=normal,
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tumor=tumor,
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qc_2=qc_2.qc,
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concordance=conpair.concordance,
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snv_result=dealwithsnvindel.snv,
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cnv_result=cnvkit.cnv,
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fusion_result=fusion.fusion,
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tmb=TMB.tmb
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}
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call hotspot{
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input:
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tumor=tumor,
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outputDir=outputDir,
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ref=ref,
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rmdupBam=markduplicates.rmdup,
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codes_dir=codes_dir
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}
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}
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#create project directory
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task create_dir{
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String outputDir
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String need_MSI_test
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String need_neoantigen_test
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command <<<
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#创建目录
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if [ ! -d ${outputDir} ];then
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mkdir ${outputDir}
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fi
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#创建qc目录
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if [ ! -d ${outputDir}/qc ];then
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mkdir ${outputDir}/qc
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fi
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#创建alignment目录
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if [ ! -d ${outputDir}/alignment ];then
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mkdir ${outputDir}/alignment
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fi
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#创建mutation目录
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if [ ! -d ${outputDir}/mutation ];then
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mkdir ${outputDir}/mutation
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fi
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#创建cnv目录
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if [ ! -d ${outputDir}/cnvkit ];then
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mkdir ${outputDir}/cnvkit
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fi
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#创建report目录
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if [ ! -d ${outputDir}/report ];then
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mkdir -p ${outputDir}/report
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fi
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#创建chemo目录
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if [ ! -d ${outputDir}/chemo ];then
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mkdir ${outputDir}/chemo
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fi
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#创建fusion目录
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if [ ! -d ${outputDir}/fusion ];then
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mkdir ${outputDir}/fusion
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fi
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#创建MSI目录
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if [ ${need_MSI_test} = "yes" ] && [ ! -d ${outputDir}/MSI ];then
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mkdir ${outputDir}/MSI
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fi
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if [ ${need_neoantigen_test} = "yes" ] && [ ! -d ${outputDir}/neoantigen/HLA ];then
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mkdir -p ${outputDir}/neoantigen/HLA
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fi
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#创建MMR目录
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if [ ! -d ${outputDir}/MMR ];then
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mkdir ${outputDir}/MMR
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fi
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##创建HRR目录
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if [ ! -d ${outputDir}/HRR ];then
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mkdir ${outputDir}/HRR
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fi
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#创建HPD目录
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if [ ! -d ${outputDir}/HPD ];then
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mkdir ${outputDir}/HPD
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fi
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>>>
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output{
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String newdir = "${outputDir}/report"
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}
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}
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#generator raw fastq to clean fastq
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task qc{
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String name
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String inputDir
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String outputDir
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command <<<
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echo processing raw reads with fastp
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fastp -i ${inputDir}/*_${name}_*1.fq.gz -o ${outputDir}/qc/${name}_clean_R1.fq.gz \
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-I ${inputDir}/*_${name}_*2.fq.gz -O ${outputDir}/qc/${name}_clean_R2.fq.gz \
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-w 10 \
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--correction \
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--overlap_len_require 10 \
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-j ${outputDir}/qc/${name}.json \
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-h ${outputDir}/qc/${name}.html --report_title ${name} -e 20
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# --adapter_sequence AAGTCGGAGGCCAAGCGGTCTTAGGAAGACAA \
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# --adapter_sequence_r2 AAGTCGGATCGTAGCCATGTCGTTCTGTGAGCCAAGGAGTTG \
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>>>
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output{
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Array[String] outputFile = [
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"${outputDir}/qc/${name}_clean_R1.fq.gz ",
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"${outputDir}/qc/${name}_clean_R2.fq.gz",
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"${outputDir}/qc/${name}.json",
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"${outputDir}/qc/${name}.html"
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]
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}
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}
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#alignment clean fastq to reference
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task alignment_bwa{
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String name
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String ref
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String outputDir
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String read1
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String read2
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command<<<
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bwa mem -R '@RG\tID:group_n\tLB:library_n\tPL:BGI\tPU:unit1\tSM:${name}' -M -t 10 ${ref} \
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${outputDir}/qc/${name}_clean_R1.fq.gz ${outputDir}/qc/${name}_clean_R2.fq.gz | \
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samtools view -@ 5 -bh -o - | samtools sort -@ 5 -o ${outputDir}/alignment/${name}.sorted.bam
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samtools index ${outputDir}/alignment/${name}.sorted.bam
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>>>
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output{
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String sorted = "${outputDir}/alignment/${name}.sorted.bam"
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}
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}
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#remove PCR duplicates
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task markduplicates{
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String name
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String ref
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String outputDir
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String sortedBam
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command<<<
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java -XX:+UseParallelGC -XX:ParallelGCThreads=2 -Xmx12G -jar $PICARD MarkDuplicates \
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I=${outputDir}/alignment/${name}.sorted.bam \
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O=${outputDir}/alignment/${name}.rmdup.bam \
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CREATE_INDEX=true \
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M=${outputDir}/alignment/${name}.rmdup.metrics.txt \
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R=${ref}
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>>>
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output{
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String rmdup = "${outputDir}/alignment/${name}.rmdup.bam"
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}
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}
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# generater mpileup file
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task generater_mpileup{
|
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String name
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String ref
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String bed
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String outputDir
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|
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String rmdupBam
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||
|
|
|
||
|
|
command<<<
|
||
|
|
|
||
|
|
samtools mpileup -Bq 20 -Q 20 -f ${ref} -l ${bed} \
|
||
|
|
${outputDir}/alignment/${name}.rmdup.bam -o ${outputDir}/alignment/${name}.pileup
|
||
|
|
|
||
|
|
>>>
|
||
|
|
|
||
|
|
output{
|
||
|
|
String pileup = "${outputDir}/alignment/${name}.pileup"
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task mutation_calling{
|
||
|
|
String codes_dir
|
||
|
|
String tumor
|
||
|
|
Array[String] pileup
|
||
|
|
String outputDir
|
||
|
|
|
||
|
|
command<<<
|
||
|
|
|
||
|
|
sh ${codes_dir}/mpileup2cns.sh ${sep=" " pileup} ${outputDir} ${tumor}
|
||
|
|
java -jar $GATK MergeVcfs -I ${outputDir}/mutation/${tumor}.snp.Somatic.hc.vcf -I ${outputDir}/mutation/${tumor}.indel.Somatic.hc.vcf \
|
||
|
|
-O ${outputDir}/mutation/${tumor}.snp.indel.Somatic.hc.vcf -D /dataseq/jmdna/database/genome/hg19/hg19.dict
|
||
|
|
java -jar $GATK MergeVcfs -I ${outputDir}/mutation/${tumor}.snp.Germline.vcf -I ${outputDir}/mutation/${tumor}.indel.Germline.vcf \
|
||
|
|
-O ${outputDir}/mutation/${tumor}.snp.indel.Germline.vcf -D /dataseq/jmdna/database/genome/hg19/hg19.dict
|
||
|
|
java -jar $GATK MergeVcfs -I ${outputDir}/mutation/${tumor}.snp.LOH.hc.vcf -I ${outputDir}/mutation/${tumor}.indel.LOH.hc.vcf \
|
||
|
|
-O ${outputDir}/mutation/${tumor}.snp.indel.LOH.hc.vcf -D /dataseq/jmdna/database/genome/hg19/hg19.dict
|
||
|
|
|
||
|
|
>>>
|
||
|
|
|
||
|
|
output{
|
||
|
|
Array[String] vcf = [
|
||
|
|
"${outputDir}/mutation/${tumor}.snp.indel.Somatic.hc.vcf",
|
||
|
|
"${outputDir}/mutation/${tumor}.snp.indel.Germline.vcf",
|
||
|
|
"${outputDir}/mutation/${tumor}.snp.indel.LOH.hc.vcf"
|
||
|
|
]
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task annovar{
|
||
|
|
String tumor
|
||
|
|
String normal
|
||
|
|
String outputDir
|
||
|
|
String ref
|
||
|
|
Array[String] vcf
|
||
|
|
command<<<
|
||
|
|
|
||
|
|
table_annovar.pl \
|
||
|
|
${outputDir}/mutation/${tumor}.snp.indel.Somatic.hc.vcf \
|
||
|
|
/dataseq/jmdna/software/annovar/humandb/ \
|
||
|
|
-buildver hg19 -nastring . -vcfinput -remove -otherinfo \
|
||
|
|
-protocol refGene,avsnp150,cosmic91,clinvar_20220320,1000g2015aug_all,1000g2015aug_eas,esp6500siv2_all,exac03nontcga,gnomad_genome,dbnsfp35c,cytoBand \
|
||
|
|
-argument '-splicing_threshold 2 -hgvs',,,,,,,,,, \
|
||
|
|
--intronhgvs 50 \
|
||
|
|
-operation g,f,f,f,f,f,f,f,f,f,r \
|
||
|
|
--outfile ${outputDir}/mutation/${tumor}.snp.indel.Somatic.anno
|
||
|
|
|
||
|
|
table_annovar.pl \
|
||
|
|
${outputDir}/mutation/${tumor}.snp.indel.Germline.vcf \
|
||
|
|
/dataseq/jmdna/software/annovar/humandb/ \
|
||
|
|
-buildver hg19 -nastring . -vcfinput -remove -otherinfo \
|
||
|
|
-protocol refGene,avsnp150,cosmic91,clinvar_20220320,1000g2015aug_all,1000g2015aug_eas,esp6500siv2_all,exac03nontcga,gnomad_genome,dbnsfp35c,cytoBand \
|
||
|
|
-argument '-splicing_threshold 2 -hgvs',,,,,,,,,, \
|
||
|
|
--intronhgvs 50 \
|
||
|
|
-operation g,f,f,f,f,f,f,f,f,f,r \
|
||
|
|
--outfile ${outputDir}/mutation/${tumor}.snp.indel.Germline.anno
|
||
|
|
|
||
|
|
table_annovar.pl \
|
||
|
|
${outputDir}/mutation/${tumor}.snp.indel.LOH.hc.vcf \
|
||
|
|
/dataseq/jmdna/software/annovar/humandb/ \
|
||
|
|
-buildver hg19 -nastring . -vcfinput -remove -otherinfo \
|
||
|
|
-protocol refGene,avsnp150,cosmic91,clinvar_20220320,1000g2015aug_all,1000g2015aug_eas,esp6500siv2_all,exac03nontcga,gnomad_genome,dbnsfp35c,cytoBand \
|
||
|
|
-argument '-splicing_threshold 2 -hgvs',,,,,,,,,, \
|
||
|
|
--intronhgvs 50 \
|
||
|
|
-operation g,f,f,f,f,f,f,f,f,f,r \
|
||
|
|
--outfile ${outputDir}/mutation/${tumor}.snp.indel.LOH.anno
|
||
|
|
|
||
|
|
java -jar /dataseq/jmdna/software/GenomeAnalysisTK.3.7.jar -T VariantAnnotator \
|
||
|
|
-R ${ref} \
|
||
|
|
-I ${outputDir}/alignment/${tumor}.rmdup.bam \
|
||
|
|
-V ${outputDir}/mutation/${tumor}.snp.indel.Somatic.hc.vcf \
|
||
|
|
-o ${outputDir}/mutation/${tumor}.TandemRepeatAnnotator.vcf \
|
||
|
|
--annotation TandemRepeatAnnotator
|
||
|
|
|
||
|
|
# -nt 10
|
||
|
|
|
||
|
|
grep -v "^##" ${outputDir}/mutation/${tumor}.TandemRepeatAnnotator.vcf | cut -f8| paste ${outputDir}/mutation/${tumor}.snp.indel.Somatic.anno.hg19_multianno.txt - >${outputDir}/mutation/${tumor}.snp.indel.Somatic.annoall.hg19_multianno.txt
|
||
|
|
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
Array[String] anno = [
|
||
|
|
"${outputDir}/mutation/${tumor}.snp.indel.Somatic.anno.hg19_multianno.txt",
|
||
|
|
"${outputDir}/mutation/${tumor}.snp.indel.Germline.anno.hg19_multianno.txt",
|
||
|
|
"${outputDir}/mutation/${tumor}.snp.indel.Somatic.annoall.hg19_multianno.txt",
|
||
|
|
]
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task dealwithsnvindel{
|
||
|
|
String codes_dir
|
||
|
|
String project
|
||
|
|
String outputDir
|
||
|
|
String tumor
|
||
|
|
Array[String] anno
|
||
|
|
String cancer
|
||
|
|
|
||
|
|
command <<<
|
||
|
|
|
||
|
|
perl ${codes_dir}/pick_variant.pl ${outputDir} ${tumor}
|
||
|
|
perl ${codes_dir}/pick_mut_splice_promoter.pl ${codes_dir} ${tumor} ${outputDir} ${project}
|
||
|
|
perl /home/jm001/test_duantao/database_update/codes/682/targetTherapy.pl ${tumor} ${outputDir} ${project} ${cancer}
|
||
|
|
perl /home/jm001/test_duantao/database_update/codes/682/germline_targetTherapy.pl ${tumor} ${outputDir} ${project} ${cancer}
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
String snv = "${outputDir}/mutation/${tumor}.snvindel.pos.txt"
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task qc_2{
|
||
|
|
String ref
|
||
|
|
String codes_dir
|
||
|
|
String name
|
||
|
|
String tumor
|
||
|
|
String normal
|
||
|
|
String bed
|
||
|
|
String outputDir
|
||
|
|
#for task chain
|
||
|
|
Array[String] anno
|
||
|
|
Boolean get_sampleinfo_local
|
||
|
|
command <<<
|
||
|
|
samtools flagstat -@ 10 ${outputDir}/alignment/${name}.rmdup.bam >${outputDir}/qc/${name}.rmdup.flagstat
|
||
|
|
samtools stats --reference ${ref} -t ${bed} -@ 10 ${outputDir}/alignment/${name}.rmdup.bam > ${outputDir}/alignment/${name}.stat
|
||
|
|
mkdir ${outputDir}/qc/${name}_bamdst
|
||
|
|
bamdst -p ${bed} -o ${outputDir}/qc/${name}_bamdst ${outputDir}/alignment/${name}.rmdup.bam
|
||
|
|
Rscript ${codes_dir}/InsertAndDepthStat.R ${outputDir}/qc/${name}_InsertAndDepthStat ${outputDir}/qc/${name}_bamdst/insertsize.plot ${outputDir}/qc/${name}_bamdst/depth_distribution.plot
|
||
|
|
# 本地获取样本信息
|
||
|
|
if [ ! ${get_sampleinfo_local} ]; then
|
||
|
|
# 获取实验样本的信息
|
||
|
|
if [ "${name}" == "${tumor}" ];then
|
||
|
|
python3 ${codes_dir}/sample_post.py -s ${tumor} -o ${outputDir} --n ${normal}
|
||
|
|
fi
|
||
|
|
else
|
||
|
|
cp /home/jm001/test_duantao/database_update/codes/temp.json ${outputDir}/qc/${normal}_post.json
|
||
|
|
fi
|
||
|
|
python3 ${codes_dir}/qc_stat.py ${outputDir}/qc/${name}.json ${outputDir}/qc/${name}_bamdst/coverage.report ${outputDir}/qc/${name}_bamdst/depth_distribution.plot ${outputDir} ${name} ${tumor}
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
String qc = "${outputDir}/qc/${name}_qc.txt"
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task fusion{
|
||
|
|
String ref
|
||
|
|
String codes_dir
|
||
|
|
String tumor
|
||
|
|
String outputDir
|
||
|
|
String cancer
|
||
|
|
String project
|
||
|
|
#for task chain
|
||
|
|
Array[String] rmdupBam
|
||
|
|
|
||
|
|
command<<<
|
||
|
|
# Extract the discordant paired-end alignments.
|
||
|
|
samtools view -b -F 1294 ${outputDir}/alignment/${tumor}.rmdup.bam > ${outputDir}/fusion/${tumor}.discordants.bam
|
||
|
|
|
||
|
|
# Extract the split-read alignments
|
||
|
|
samtools view -h ${outputDir}/alignment/${tumor}.rmdup.bam \
|
||
|
|
| /dataseq/jmdna/software/lumpy-sv/scripts/extractSplitReads_BwaMem -i stdin \
|
||
|
|
| samtools view -Sb - \
|
||
|
|
> ${outputDir}/fusion/${tumor}.splitters.bam
|
||
|
|
|
||
|
|
lumpyexpress \
|
||
|
|
-B ${outputDir}/alignment/${tumor}.rmdup.bam \
|
||
|
|
-S ${outputDir}/fusion/${tumor}.splitters.bam \
|
||
|
|
-D ${outputDir}/fusion/${tumor}.discordants.bam \
|
||
|
|
-o ${outputDir}/fusion/${tumor}.fusion.vcf
|
||
|
|
|
||
|
|
perl ${codes_dir}/fusion.filter.pl ${outputDir}/fusion/${tumor}.fusion.vcf ${outputDir}/fusion/${tumor}.fusion.filter.vcf
|
||
|
|
|
||
|
|
svtyper \
|
||
|
|
-B ${outputDir}/alignment/${tumor}.rmdup.bam \
|
||
|
|
-i ${outputDir}/fusion/${tumor}.fusion.filter.vcf \
|
||
|
|
-T ${ref} \
|
||
|
|
-o ${outputDir}/fusion/${tumor}.fusion.gt.vcf
|
||
|
|
|
||
|
|
table_annovar.pl \
|
||
|
|
${outputDir}/fusion/${tumor}.fusion.gt.vcf \
|
||
|
|
/dataseq/jmdna/software/annovar/humandb/ \
|
||
|
|
-buildver hg19 -nastring . -vcfinput -remove -otherinfo \
|
||
|
|
-protocol refGene \
|
||
|
|
-operation g \
|
||
|
|
--outfile ${outputDir}/fusion/${tumor}.fusion
|
||
|
|
|
||
|
|
perl ${codes_dir}/fusion.reanno.pl ${outputDir}/qc/${tumor}_bamdst/depth.tsv.gz ${outputDir} ${tumor}
|
||
|
|
perl /home/jm001/test_duantao/database_update/codes/682/fusion_targetTherapy.pl ${codes_dir} ${tumor} ${outputDir} ${project} ${cancer}
|
||
|
|
|
||
|
|
>>>
|
||
|
|
|
||
|
|
output{
|
||
|
|
String fusion = "${outputDir}/fusion/${tumor}.fusion.pos.txt"
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task chemoTherapy{
|
||
|
|
String codes_dir
|
||
|
|
String normal
|
||
|
|
String tumor
|
||
|
|
String outputDir
|
||
|
|
String ref
|
||
|
|
String project
|
||
|
|
Array[String] rmdupBam
|
||
|
|
command <<<
|
||
|
|
# perl ${codes_dir}/chemo/singlecancer_chemo_2.pl ${codes_dir} ${outputDir} ${normal} ${ref} ${project}
|
||
|
|
${codes_dir}/chemo/chemo_panel.py -p ${project} -o ${outputDir} --n ${normal}
|
||
|
|
>>>
|
||
|
|
}
|
||
|
|
|
||
|
|
task hereditary{
|
||
|
|
String codes_dir
|
||
|
|
String tumor
|
||
|
|
String outputDir
|
||
|
|
String project
|
||
|
|
String snv
|
||
|
|
command <<<
|
||
|
|
${codes_dir}/hereditary/hereditary.py -p ${project} -o ${outputDir} --n ${tumor}
|
||
|
|
>>>
|
||
|
|
}
|
||
|
|
|
||
|
|
task conpair{
|
||
|
|
String name
|
||
|
|
String tumor
|
||
|
|
String normal
|
||
|
|
String outputDir
|
||
|
|
String codes_dir
|
||
|
|
String ref
|
||
|
|
#for task chain
|
||
|
|
Array[String] rmdupBam
|
||
|
|
command <<<
|
||
|
|
|
||
|
|
python3 /dataseq/jmdna/software/Conpair-master/scripts/run_gatk_pileup_for_sample.py \
|
||
|
|
-M /dataseq/jmdna/software/Conpair-master/data/markers/GRCh37.autosomes.phase3_shapeit2_mvncall_integrated.20130502.SNV.genotype.sselect_v4_MAF_0.4_LD_0.8.bed \
|
||
|
|
-B ${outputDir}/alignment/${normal}.rmdup.bam \
|
||
|
|
-O ${outputDir}/alignment/${normal}.gatk.mpileup \
|
||
|
|
-R ${ref} \
|
||
|
|
-G /dataseq/jmdna/software/GenomeAnalysisTK.3.7.jar
|
||
|
|
|
||
|
|
python3 /dataseq/jmdna/software/Conpair-master/scripts/run_gatk_pileup_for_sample.py \
|
||
|
|
-M /dataseq/jmdna/software/Conpair-master/data/markers/GRCh37.autosomes.phase3_shapeit2_mvncall_integrated.20130502.SNV.genotype.sselect_v4_MAF_0.4_LD_0.8.bed \
|
||
|
|
-B ${outputDir}/alignment/${tumor}.rmdup.bam \
|
||
|
|
-O ${outputDir}/alignment/${tumor}.gatk.mpileup \
|
||
|
|
-R ${ref} \
|
||
|
|
-G /dataseq/jmdna/software/GenomeAnalysisTK.3.7.jar
|
||
|
|
|
||
|
|
sed -i 's/^chr//g' ${outputDir}/alignment/${normal}.gatk.mpileup
|
||
|
|
sed -i 's/^chr//g' ${outputDir}/alignment/${tumor}.gatk.mpileup
|
||
|
|
|
||
|
|
|
||
|
|
python3 /dataseq/jmdna/software/Conpair-master/scripts/verify_concordance.py \
|
||
|
|
-T ${outputDir}/alignment/${tumor}.gatk.mpileup \
|
||
|
|
-N ${outputDir}/alignment/${normal}.gatk.mpileup \
|
||
|
|
-H \
|
||
|
|
-O ${outputDir}/qc/${name}_concordance.txt
|
||
|
|
|
||
|
|
python3 /dataseq/jmdna/software/Conpair-master/scripts/estimate_tumor_normal_contamination.py \
|
||
|
|
-T ${outputDir}/alignment/${tumor}.gatk.mpileup \
|
||
|
|
-N ${outputDir}/alignment/${normal}.gatk.mpileup \
|
||
|
|
-O ${outputDir}/qc/${name}_contamination.txt
|
||
|
|
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
Array[String] concordance = [
|
||
|
|
"${outputDir}/report/qc/${name}_concordance.txt",
|
||
|
|
"${outputDir}/report/qc/${name}_contamination.txt",
|
||
|
|
]
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task MSI{
|
||
|
|
String bed
|
||
|
|
String tumor
|
||
|
|
String normal
|
||
|
|
String outputDir
|
||
|
|
Array[String] rmdupBam
|
||
|
|
|
||
|
|
command <<<
|
||
|
|
|
||
|
|
msisensor2 msi -d /dataseq/jmdna/software/msisensor2/hg19.microsatellites.list -n ${outputDir}/alignment/${normal}.rmdup.bam -t ${outputDir}/alignment/${tumor}.rmdup.bam -e ${bed} -b 10 -o ${outputDir}/MSI/${tumor}.msi
|
||
|
|
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
String target="${outputDir}/MSI/${tumor}.msi"
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task MMR{
|
||
|
|
String codes_dir
|
||
|
|
String tumor
|
||
|
|
String outputDir
|
||
|
|
String snv
|
||
|
|
command <<<
|
||
|
|
perl ${codes_dir}/mmr_controlsample.pl ${outputDir} ${tumor}
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
String mmr_out = "${outputDir}/MMR/${tumor}_mmr.txt"
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task HRR{
|
||
|
|
String codes_dir
|
||
|
|
String tumor
|
||
|
|
String outputDir
|
||
|
|
String snv
|
||
|
|
command <<<
|
||
|
|
perl ${codes_dir}/hrr_controlsample_tissue.pl ${outputDir} ${tumor}
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
String mmr_out = "${outputDir}/HRR/${tumor}_hrr.txt"
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task TMB{
|
||
|
|
String codes_dir
|
||
|
|
String tumor
|
||
|
|
String outputDir
|
||
|
|
Array[String] anno
|
||
|
|
Array[String] concordance
|
||
|
|
String snv_result
|
||
|
|
String cnv_result
|
||
|
|
String fusion_result
|
||
|
|
|
||
|
|
|
||
|
|
|
||
|
|
|
||
|
|
|
||
|
|
command <<<
|
||
|
|
perl ${codes_dir}/tmb.pl ${outputDir} ${tumor}
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
String tmb="${outputDir}/mutation/${tumor}.tmb.txt"
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task cnvkit{
|
||
|
|
String ref
|
||
|
|
String bed
|
||
|
|
String name
|
||
|
|
String tumor
|
||
|
|
String normal
|
||
|
|
String outputDir
|
||
|
|
String cancer
|
||
|
|
String accessBed
|
||
|
|
String annotateGene
|
||
|
|
String codes_dir
|
||
|
|
String project
|
||
|
|
#for task chain
|
||
|
|
Array[String] rmdupBam
|
||
|
|
String purity
|
||
|
|
|
||
|
|
command <<<
|
||
|
|
|
||
|
|
echo run cnvkit batch to processing cnv calling
|
||
|
|
cnvkit.py batch \
|
||
|
|
${outputDir}/alignment/${tumor}.rmdup.bam \
|
||
|
|
--normal ${outputDir}/alignment/${normal}.rmdup.bam \
|
||
|
|
--targets ${bed} \
|
||
|
|
--fasta ${ref} \
|
||
|
|
--access ${accessBed} \
|
||
|
|
--output-reference ${outputDir}/cnvkit/${normal}_reference.cnn \
|
||
|
|
--annotate ${annotateGene} \
|
||
|
|
--drop-low-coverage --scatter --output-dir ${outputDir}/cnvkit
|
||
|
|
|
||
|
|
cnvkit.py scatter \
|
||
|
|
${outputDir}/cnvkit/${tumor}.rmdup.cnr -s ${outputDir}/cnvkit/${tumor}.rmdup.cns \
|
||
|
|
--y-max 3 --y-min -3 \
|
||
|
|
--title ${tumor}.cns \
|
||
|
|
-o ${outputDir}/cnvkit/${tumor}.cnv.png
|
||
|
|
|
||
|
|
if [ -e "${outputDir}/qc/sequenza/${name}_confints_CP.txt" ]; then
|
||
|
|
# absolute copy number
|
||
|
|
cnvkit.py call \
|
||
|
|
-m clonal \
|
||
|
|
${outputDir}/cnvkit/${tumor}.rmdup.cns \
|
||
|
|
-y \
|
||
|
|
--purity `head -n2 ${outputDir}/qc/sequenza/${name}_confints_CP.txt |tail -n1|cut -f1` \
|
||
|
|
--drop-low-coverage \
|
||
|
|
--filter ampdel \
|
||
|
|
-o ${outputDir}/cnvkit/${tumor}.rmdup.cns.cn.hc
|
||
|
|
fi
|
||
|
|
perl ${codes_dir}/log2_cn.pl ${outputDir}/cnvkit/${tumor}.rmdup.cns ${outputDir}/cnvkit/${tumor}.rmdup.cns.cn
|
||
|
|
perl /home/jm001/test_duantao/database_update/codes/682/cnv_targetTherapy.pl ${codes_dir} ${tumor} ${outputDir} ${project} ${cancer}
|
||
|
|
>>>
|
||
|
|
|
||
|
|
output{
|
||
|
|
String cnv = "${outputDir}/cnvkit/${tumor}.rmdup.cns"
|
||
|
|
}
|
||
|
|
|
||
|
|
}
|
||
|
|
|
||
|
|
task tumor_content{
|
||
|
|
String name
|
||
|
|
String ref
|
||
|
|
String tumor
|
||
|
|
String normal
|
||
|
|
String outputDir
|
||
|
|
String gc_wiggle
|
||
|
|
String codes_dir
|
||
|
|
#for task chain
|
||
|
|
Array[String] pileup
|
||
|
|
|
||
|
|
command <<<
|
||
|
|
|
||
|
|
sequenza-utils bam2seqz -p -gc ${gc_wiggle} \
|
||
|
|
-F ${ref} \
|
||
|
|
-n ${outputDir}/alignment/${normal}.pileup \
|
||
|
|
-t ${outputDir}/alignment/${tumor}.pileup | gzip >${outputDir}/qc/target_${name}.200base.seqz.gz
|
||
|
|
sequenza-utils seqz_binning -w 200 -s ${outputDir}/qc/target_${name}.200base.seqz.gz | gzip > ${outputDir}/qc/target_${name}.200base.small.seqz.gz
|
||
|
|
|
||
|
|
Rscript ${codes_dir}/sequenza.R ${name} ${outputDir}/qc/target_${name}.200base.small.seqz.gz ${outputDir}/qc/sequenza || echo "sequenza failed!"
|
||
|
|
|
||
|
|
|
||
|
|
# Rscript ${codes_dir}/qc_stat.R ${outputDir} ${name}
|
||
|
|
# cp ${outputDir}/qc/${name}_qcstat.txt ${outputDir}/report/qc/${name}_qcstat.txt
|
||
|
|
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
String purity = "${outputDir}/qc/sequenza/${name}_CP_contours.pdf"
|
||
|
|
}
|
||
|
|
|
||
|
|
}
|
||
|
|
|
||
|
|
task HLA{
|
||
|
|
String inputDir
|
||
|
|
String outputDir
|
||
|
|
String normal
|
||
|
|
String newdir
|
||
|
|
command <<<
|
||
|
|
|
||
|
|
razers3 -tc 10 -i 95 -m 1 -dr 0 -o ${outputDir}/neoantigen/HLA/fished_1.bam /dataseq/jmdna/software/OptiType-1.3.5/data/hla_reference_dna.fasta ${inputDir}/*_${normal}_*1.fq.gz
|
||
|
|
samtools bam2fq ${outputDir}/neoantigen/HLA/fished_1.bam > ${outputDir}/neoantigen/HLA/${normal}_1_fished.fastq
|
||
|
|
rm ${outputDir}/neoantigen/HLA/fished_1.bam
|
||
|
|
razers3 -tc 10 -i 95 -m 1 -dr 0 -o ${outputDir}/neoantigen/HLA/fished_2.bam /dataseq/jmdna/software/OptiType-1.3.5/data/hla_reference_dna.fasta ${inputDir}/*_${normal}_*2.fq.gz
|
||
|
|
samtools bam2fq ${outputDir}/neoantigen/HLA/fished_2.bam > ${outputDir}/neoantigen/HLA/${normal}_2_fished.fastq
|
||
|
|
rm ${outputDir}/neoantigen/HLA/fished_2.bam
|
||
|
|
/dataseq/jmdna/software/OptiType-1.3.5/OptiTypePipeline.py -i ${outputDir}/neoantigen/HLA/${normal}_1_fished.fastq ${outputDir}/neoantigen/HLA/${normal}_2_fished.fastq --dna -v --prefix ${normal} -o ${outputDir}/neoantigen/HLA/
|
||
|
|
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
String hla = "${outputDir}/neoantigen/HLA/${normal}_result.tsv"
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task neoantigen{
|
||
|
|
String codes_dir
|
||
|
|
String outputDir
|
||
|
|
String name
|
||
|
|
String normal
|
||
|
|
String tumor
|
||
|
|
String hla
|
||
|
|
Array[String] vcf
|
||
|
|
command <<<
|
||
|
|
sh /home/jm001/test_duantao/database_update/test_project/20230814_test/predict_neoantigen.sh ${outputDir} ${name} ${tumor} ${codes_dir}
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
String neoantigen = "${outputDir}/neoantigen/MHC_Class_I/${tumor}.all_epitopes.netchop.txt"
|
||
|
|
}
|
||
|
|
}
|
||
|
|
|
||
|
|
task auto_report{
|
||
|
|
String cancer
|
||
|
|
String codes_dir
|
||
|
|
String outputDir
|
||
|
|
String name
|
||
|
|
String normal
|
||
|
|
String tumor
|
||
|
|
Array[String] qc_2
|
||
|
|
Array[String] concordance
|
||
|
|
# String purity
|
||
|
|
String snv_result
|
||
|
|
String cnv_result
|
||
|
|
String fusion_result
|
||
|
|
String tmb
|
||
|
|
|
||
|
|
command <<<
|
||
|
|
|
||
|
|
perl /home/jm001/test_duantao/database_update/codes/682/indication.pl ${outputDir} ${cancer}
|
||
|
|
python3 ${codes_dir}/qc_check_tissue.py ${outputDir} ${tumor} ${normal} ${name}
|
||
|
|
python3 ${codes_dir}/drug_dedup.py ${outputDir} ${tumor}
|
||
|
|
perl ${codes_dir}/hpd.pl ${outputDir} ${tumor}
|
||
|
|
python3 ${codes_dir}/682gene_tissue_control_report.py ${outputDir} ${tumor} ${normal} ${cancer}
|
||
|
|
python3 ${codes_dir}/wdl_check.py -o ${outputDir}
|
||
|
|
perl ${codes_dir}/file_format_change.pl ${outputDir} ${tumor}
|
||
|
|
# cp ${outputDir}/qc/sequenza/${name}_confints_CP.txt ${outputDir}/report/${name}_confints_CP.txt
|
||
|
|
cp ${outputDir}/qc/qc_fail.txt ${outputDir}/report/qc_fail.txt
|
||
|
|
cp ${outputDir}/qc/${tumor}_qc.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/cnvkit/${tumor}.rmdup.cns.cn ${outputDir}/report/
|
||
|
|
cp ${outputDir}/cnvkit/${tumor}.cnv.png ${outputDir}/report/
|
||
|
|
cp ${outputDir}/fusion/${tumor}.fuison.vus.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/fusion/${tumor}.fusion.reanno.vcf ${outputDir}/report/
|
||
|
|
# cp ${outputDir}/mutation/*filter* ${outputDir}/report/
|
||
|
|
cp ${outputDir}/mutation/${tumor}.snvindel.Germline.target.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/mutation/${tumor}.nontarget.vus.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/mutation/${tumor}.snvindel.vus.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/mutation/${tumor}.target.splicing.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/mutation/${tumor}.target.promoter.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/mutation/${tumor}.tmb.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/MMR/${tumor}.mmr.pre.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/HRR/${tumor}.hrr.pre.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/HPD/${tumor}.hpd.pre.txt ${outputDir}/report/
|
||
|
|
cp ${outputDir}/hereditary/${tumor}.hereditary.pre.txt ${outputDir}/report/
|
||
|
|
python3 ${codes_dir}/txt_xlsx.py ${outputDir} ${tumor}
|
||
|
|
>>>
|
||
|
|
}
|
||
|
|
|
||
|
|
# hotspot
|
||
|
|
task hotspot{
|
||
|
|
String tumor
|
||
|
|
String outputDir
|
||
|
|
String ref
|
||
|
|
Array[String] rmdupBam
|
||
|
|
String codes_dir
|
||
|
|
command<<<
|
||
|
|
|
||
|
|
mkdir -p ${outputDir}/mutation/hotspot
|
||
|
|
samtools mpileup -Bq 20 -Q 20 -f ${ref} -l ${codes_dir}/hotspot.bed ${outputDir}/alignment/${tumor}.rmdup.bam -o ${outputDir}/mutation/hotspot/${tumor}.hotspot.pileup
|
||
|
|
java -jar $VARSCAN mpileup2cns ${outputDir}/mutation/hotspot/${tumor}.hotspot.pileup --min-var-freq 0.005 --min-avg-qual 20 --output-vcf 1 --variants 1 --p-value 0.99 --min-reads2 2 --strand-filter 0 > ${outputDir}/mutation/hotspot/${tumor}.hotspot.L.snp.indel.vcf
|
||
|
|
java -jar $VARSCAN mpileup2cns ${outputDir}/mutation/hotspot/${tumor}.hotspot.pileup --min-var-freq 0.01 --min-avg-qual 20 --output-vcf 1 --variants 1 --p-value 0.05 --min-reads2 3 --strand-filter 1 > ${outputDir}/mutation/hotspot/${tumor}.hotspot.H.snp.indel.vcf
|
||
|
|
perl ${codes_dir}/hotspot.hvl.pl ${outputDir} ${tumor}
|
||
|
|
|
||
|
|
if [ -e "${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.vcf" ]; then
|
||
|
|
table_annovar.pl \
|
||
|
|
${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.vcf \
|
||
|
|
/dataseq/jmdna/software/annovar/humandb/ \
|
||
|
|
-buildver hg19 -nastring . -vcfinput -remove -otherinfo \
|
||
|
|
-protocol refGene \
|
||
|
|
-argument '-hgvs' \
|
||
|
|
-operation g \
|
||
|
|
--outfile ${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.anno
|
||
|
|
|
||
|
|
|
||
|
|
# java -jar /dataseq/jmdna/software/GenomeAnalysisTK.3.7.jar -T VariantAnnotator \
|
||
|
|
# -R ${ref} \
|
||
|
|
# -I ${outputDir}/alignment/${tumor}.rmdup.bam \
|
||
|
|
# -V ${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.vcf \
|
||
|
|
# -o ${outputDir}/mutation/hotspot/${tumor}.hotspot.TandemRepeatAnnotator.vcf \
|
||
|
|
# --annotation TandemRepeatAnnotator
|
||
|
|
|
||
|
|
# grep -v "^##" ${outputDir}/mutation/hotspot/${tumor}.hotspot.TandemRepeatAnnotator.vcf | cut -f8| paste ${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.anno.hg19_multianno.txt - >${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.Somatic.annoall.hg19_multianno.txt
|
||
|
|
# cp ${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.anno.hg19_multianno.txt ${outputDir}/report/${tumor}.hotspot.snp.indel.anno.hg19_multianno.txt
|
||
|
|
|
||
|
|
perl ${codes_dir}/hotspot.filter.pl ${outputDir} ${tumor}
|
||
|
|
fi
|
||
|
|
|
||
|
|
>>>
|
||
|
|
output{
|
||
|
|
String hotspot = "${outputDir}/mutation/hotspot/${tumor}.hotspot.H.snp.indel.vcf"
|
||
|
|
}
|
||
|
|
}
|