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version = "0.26.0"
container_url = "lbmc/kb:${version}"
params.index_fasta = ""
params.index_fasta_out = ""
workflow index_fasta {
take:
fasta
gtf
main:
emit:
index = index_default.out.index
t2g = index_default.out.t2g
report = index_default.out.report
}
process tr2g {
// create transcript to gene table from gtf if no transcript to gene file is provided
label "big_mem_mono_cpus"
tag "$file_id"
if (params.index_fasta_out != "") {
publishDir "results/${params.index_fasta_out}", mode: 'copy'
}
input:
tuple val(file_id), path(gtf)
output:
tuple val(file_id), path("t2g.txt"), emit: t2g
script:
"""
"""
}
process index_default {
container = "${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(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.fa ${fasta} ${gtf} > ${fasta.simpleName}_kb_index_report.txt
include { split } from "./../flexi_splitter/main.nf"
params.kb_protocol = "10x_v3"
params.count = ""
params.count_out = ""
workflow count {
take:
index
fastq
transcript_to_gene
whitelist
main:
whitelist
.ifEmpty(["NO WHITELIST", 0])
.set{ whitelist_optional }
switch(params.kb_protocol) {
case "marsseq":
kb_marseq(index.collect(), split.out.fastq, transcript_to_gene.collect(), whitelist_optional.collect())
kb_marseq.out.counts.set{res_counts}
kb_marseq.out.report.set{res_report}
break;
default:
kb_default(index.collect(), fastq, transcript_to_gene.collect(), whitelist_optional.collect())
kb_default.out.counts.set{res_counts}
kb_default.out.report.set{res_report}
break;
}
emit:
counts = res_counts
report = res_report
}
process kb_default {
container = "${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"){
}
if (reads.size() == 2)
"""
mkdir ${file_prefix}
kb count -t ${task.cpus} \
-i ${index} \
-g ${transcript_to_gene} \
${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"){
}
if (reads.size() == 2)
"""
mkdir ${file_prefix}
kb count -t ${task.cpus} \
-i ${index} \
-g ${transcript_to_gene} \
${reads[0]} ${reads[1]} > ${file_prefix}_kb_mapping_report.txt
else
"""
mkdir ${file_prefix}
kb count -t ${task.cpus} \
-m ${kb_memory} \
-i ${index} \
-g ${transcript_to_gene} \
${reads} > ${file_prefix}_kb_mapping_report.txt
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}
// ************************** velocity workflow **************************
workflow index_fasta_velocity {
take:
fasta
gtf
main:
tr2g(gtf)
index_fasta_velocity_default(fasta, gtf, tr2g.out.t2g)
emit:
index = index_fasta_velocity_default.out.index
t2g = index_fasta_velocity_default.out.t2g
report = index_fasta_velocity_default.out.report
}
process index_fasta_velocity_default {
container = "${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(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}"), path("cdna_t2c.txt"), path("intron_t2c.txt"), 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.fa -f2 intron.fa -c1 cdna_t2c.txt -c2 intron_t2c.txt --workflow lamanno \
${fasta} ${gtf} > ${fasta.simpleName}_kb_index_report.txt
"""
}
params.count_velocity = ""
params.count_velocity_out = ""
workflow count_velocity {
take:
index
fastq
transcript_to_gene
whitelist
config
main:
whitelist
.ifEmpty(["NO WHITELIST", 0])
.set{ whitelist_optional }
switch(params.kb_protocol) {
case "marsseq":
split(fastq, config)
velocity_marseq(index.collect(), split.out.fastq, transcript_to_gene.collect(), whitelist_optional.collect())
velocity_marseq.out.counts.set{res_counts}
velocity_marseq.out.report.set{res_report}
break;
default:
velocity_default(index.collect(), fastq, transcript_to_gene.collect(), whitelist_optional.collect())
velocity_default.out.counts.set{res_counts}
velocity_default.out.report.set{res_report}
break;
}
emit:
counts = res_counts
report = res_report
}
process velocity_default {
container = "${container_url}"
label "big_mem_multi_cpus"
tag "$file_prefix"
if (params.count_velocity_out != "") {
publishDir "results/${params.count_velocity_out}", mode: 'copy'
}
input:
tuple val(index_id), path(index)
tuple val(file_id), path(reads)
tuple val(t2g_id), path(transcript_to_gene), path(cdna_t2g), path(intron_t2g)
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}
-m ${kb_memory} \
-i ${index} \
-g ${transcript_to_gene} \
-o ${file_prefix} \
-c1 ${cdna_t2g} \
-c2 ${intron_t2g} \
--lamanno \
${whitelist_param} \
-x 10XV3 \
${params.count} \
${reads[0]} ${reads[1]} > ${file_prefix}_kb_mapping_report.txt
cp ${transcript_to_gene} ${file_prefix}/
cp ${cdna_t2g} ${file_prefix}/
cp ${intron_t2g} ${file_prefix}/
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"""
}
process velocity_marseq {
// With the MARS-Seq protocol, we have:
// on the read 1: 4 nt of bc plate
// on the read 2: 6 nt of bc cell, and 8 nt of UMI
// this process expect that the bc plate is removed from the read 1
container = "${container_url}"
label "big_mem_multi_cpus"
tag "$file_prefix"
if (params.count_velocity_out != "") {
publishDir "results/${params.count_velocity_out}", mode: 'copy'
}
input:
tuple val(index_id), path(index)
tuple val(file_id), path(reads)
tuple val(t2g_id), path(transcript_to_gene), path(cdna_t2g), path(intron_t2g)
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} \
-o ${file_prefix} \
-c1 ${cdna_t2g} \
-c2 ${intron_t2g} \
--lamanno \
${whitelist_param} \
${params.count} \
-x 1,0,6:1,6,14:0,0,0 \
${reads[0]} ${reads[1]} > ${file_prefix}_kb_mapping_report.txt
cp ${transcript_to_gene} ${file_prefix}/
cp ${cdna_t2g} ${file_prefix}/
cp ${intron_t2g} ${file_prefix}/
"""
else
"""
mkdir ${file_prefix}
kb count -t ${task.cpus} \
-m ${kb_memory} \
-i ${index} \
-g ${transcript_to_gene} \
-o ${file_prefix} \
-c1 ${cdna_t2g} \
-c2 ${intron_t2g} \
--lamanno \
${whitelist_param} \
${params.count} \
-x 1,0,6:1,6,14:0,0,0 \
${reads} > ${file_prefix}_kb_mapping_report.txt
cp ${transcript_to_gene} ${file_prefix}/
cp ${cdna_t2g} ${file_prefix}/
cp ${intron_t2g} ${file_prefix}/