version = "0.26.0" container_url = "lbmc/kb:${version}" params.index_fasta = "" params.index_fasta_out = "" workflow index_fasta { take: fasta cdna gtf transcript_to_gene main: transcript_to_gene .ifEmpty(["NO T2G", ""]) .set{ transcript_to_gene_optional } tr2g(gtf, transcript_to_gene_optional) index_default(fasta, cdna, gtf, tr2g.out.t2g) emit: index = index_default.out.index t2g = index_default.out.t2g report = index_default.out.report } process tr2g { // create transcript to gene table from gtf if no transcript to gene file is provided container = "${container_url}" label "big_mem_mono_cpus" tag "$file_id" if (params.index_fasta_out != "") { publishDir "results/${params.index_fasta_out}", mode: 'copy' } input: tuple val(file_id), path(gtf) tuple val(t2g_id), file(transcript_to_gene) output: tuple val(file_id), path("t2g.txt"), emit: t2g script: if (t2g_id == "NO T2G") """ t2g.py --gtf ${gtf} """ else """ mv ${transcript_to_gene} t2g.txt """ } process index_default { container = "${container_url}" label "big_mem_mono_cpus" tag "$file_id" if (params.index_fasta_out != "") { publishDir "results/${params.index_fasta_out}", mode: 'copy' } input: tuple val(file_id), path(fasta) tuple val(cdna_id), path(cdna) tuple val(gtf_id), path(gtf) tuple val(t2g_id), path(transcript_to_gene) output: tuple val(file_id), path("*.idx"), emit: index tuple val(t2g_id), path("${transcript_to_gene}"), emit: t2g tuple val(file_id), path("*_report.txt"), emit: report script: """ kb ref \ -i ${fasta.simpleName}.idx \ -g ${transcript_to_gene} \ ${params.index_fasta} \ -f1 ${cdna} ${fasta} ${gtf} > ${fasta.simpleName}_kb_index_report.txt """ } include { split } from "./../flexi_splitter/main.nf" params.kb_protocol = "10x_v3" params.count = "" params.count_out = "" workflow count { take: index fastq transcript_to_gene whitelist config main: whitelist .ifEmpty(["NO WHITELIST", 0]) .set{ whitelist_optional } switch(params.kb_protocol) { case "marsseq": split(fastq, config) kb_marseq(index.collect(), split.out.fastq.view(), transcript_to_gene, whitelist_optional) kb_marseq.out.counts.set{res_counts} kb_marseq.out.report.set{res_report} break; default: kb_default(index, fastq, transcript_to_gene, whitelist_optional) kb_default.out.counts.set{res_counts} kb_default.out.report.set{res_report} break; } emit: counts = res_counts report = res_report } process kb_default { container = "${container_url}" label "big_mem_multi_cpus" tag "$file_prefix" if (params.count_out != "") { publishDir "results/${params.count_out}", mode: 'copy' } input: tuple val(index_id), path(index) tuple val(file_id), path(reads) tuple val(t2g_id), path(transcript_to_gene) tuple val(whitelist_id), path(whitelist) output: tuple val(file_id), path("${file_prefix}"), emit: counts tuple val(file_id), path("*_report.txt"), emit: report script: def kb_memory = "${task.memory}" - ~/GB/ if (file_id instanceof List){ file_prefix = file_id[0] } else { file_prefix = file_id } def whitelist_param = "" if (whitelist_id != "NO WHITELIST"){ whitelist_param = "-w ${whitelist}" } if (reads.size() == 2) """ mkdir ${file_prefix} kb count -t ${task.cpus} \ -m ${kb_memory} \ -i ${index} \ -g ${transcript_to_gene} \ ${whitelist_param} \ -x 10XV3 ${params.count} \ -o ${file_prefix} \ ${reads[0]} ${reads[1]} &> ${file_prefix}_kb_mapping_report.txt """ } process kb_marseq { // With the MARS-Seq protocol, we have: // on the read 1: 4 nt of bc plate // on the read 2: 6 nt of bc cell, and 8 nt of UMI // this process expect that the bc plate is removed from the read 1 container = "${container_url}" label "big_mem_multi_cpus" tag "$file_prefix" if (params.count_out != "") { publishDir "results/${params.count_out}", mode: 'copy' } input: tuple val(index_id), path(index) tuple val(file_id), path(reads) tuple val(t2g_id), path(transcript_to_gene) tuple val(whitelist_id), path(whitelist) output: tuple val(file_id), path("${file_prefix}"), emit: counts tuple val(file_id), path("*_report.txt"), emit: report script: def kb_memory = "${task.memory}" - ~/GB/ if (file_id instanceof List){ file_prefix = file_id[0] } else { file_prefix = file_id } def whitelist_param = "" if (whitelist_id != "NO WHITELIST"){ whitelist_param = "-w ${whitelist}" } if (reads.size() == 2) """ mkdir ${file_prefix} kb count -t ${task.cpus} \ -m ${kb_memory} \ -i ${index} \ -g ${transcript_to_gene} \ ${whitelist_param} \ ${params.count} \ -x 1,0,6:1,6,14:1,14,0 -o ${file_prefix} \ ${reads[0]} ${reads[1]} &> ${file_prefix}_kb_mapping_report.txt """ }