;; fastq sensitive short read SARS-CoV-2 mutation calling (use-modules (ccwl ccwl)) (define fastq-1 (input "fastq_1")) (define fastq-2 (input "fastq_2")) (define bowtie2-index (input "bowtie2_index" #:other '((secondary-files . #(".1.bt2" ".2.bt2" ".3.bt2" ".4.bt2" ".rev.1.bt2" ".rev.2.bt2"))))) (define bwa-mem-index (input "bwa_mem_index" #:other '((secondary-files . #(".amb" ".ann" ".bwt" ".pac" ".sa"))))) (define reference (input "reference" #:other '((secondary-files . #(".fai"))))) (define (bam2vcf prefix) (workflow (string-append prefix "_bam2vcf") (list (pipeline "samtools" (list (command "samtools_view" (list "samtools" "view" "-h" "-b" "-F" "4" (input (string-append prefix "_input")))) (command "samtools_sort" (list "samtools" "sort" "-n")) (command "samtools_fixmate" (list "samtools" "fixmate" "-m" "-" "-")) (command "samtools_sort2" (list "samtools" "sort" "-")) (command "samtools_markdup" (list "samtools" "markdup" "-" "-") #:other '((stdout . "dedup.bam")))) (list (output "dedup_bam" #:type 'File #:source "samtools_markdup/stdout"))) (command "samtools_index" (list "samtools" "index" (input "dedup_bam")) #:outputs (list (output "indexed_dedup_bam" #:type 'File #:binding '((glob . "$(inputs.dedup_bam.basename)")) #:other '((secondary-files . #(".bai"))))) #:other `((hints (Initial-work-dir-requirement (listing . #("$(inputs.dedup_bam)")))))) (pipeline "freebayes" (list (command "freebayes" (list "freebayes" "--ploidy" "1" "--bam" (input "indexed_dedup_bam" #:other '((secondary-files . #("bai")))) "-f" reference)) (command "bcftools_view" (list "bcftools" "view" "-i" "QUAL > 10" "-Oz") #:other `((stdout . ,(string-append prefix "_output.vcf.gz"))))) (list (output "vcf" #:type 'File #:source "bcftools_view/stdout"))) (command "tabix" #:outputs (list (output "indexed_vcf" #:type 'File (list "tabix" (input "vcf")) #:binding '((glob . "$(inputs.vcf.basename)")) #:other '((secondary-files . #(".tbi"))))) #:other `((hints (Initial-work-dir-requirement (listing . #("$(inputs.vcf)"))))))) (list (output (string-append prefix "_output") #:type 'File #:source "tabix/indexed_vcf")))) (define variant-calling (workflow "variant-calling" (list (command "bowtie" (list "bowtie2" "-p" "16" "-x" bowtie2-index "-1" fastq-1 "-2" fastq-2) #:outputs (list (output "bowtie_input" #:type 'stdout))) (bam2vcf "bowtie") (command "bwa_mem" (list "bwa" "mem" "-K" "100000000" "-v" "3" "-t" "16" "-Y" bwa-mem-index fastq-1 fastq-2) #:outputs (list (output "bwa_mem_input" #:type 'stdout))) (bam2vcf "bwa_mem")) (list (output "bowtie_output" #:type 'File #:source "bowtie_bam2vcf/bowtie_output") (output "bwa_mem_output" #:type 'File #:source "bwa_mem_bam2vcf/bwa_mem_output")))) (write-cwl variant-calling "workflows/variant-calling-bowtie-bwa-mem.cwl")