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vvanoost
nextflow
Commits
30e9022a
Commit
30e9022a
authored
6 years ago
by
vvanoost
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CCA removal from mature tRNA nf
parent
15bc6ba1
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src/RNAseq_sen1D_bowtie2_SE_CCA.nf
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src/RNAseq_sen1D_bowtie2_SE_CCA.nf
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30e9022a
/*
* cutadapt :
* Imputs : fastq files
* Output : fastq files
*/
/* Small RNA-seq Illumina adaptor removal NEXTflex Small RNA Seq Kit v3 */
/*
* for single-end data
*/
params
.
fastq
=
"$baseDir/data/fastq_SE/*.fastq.gz"
log
.
info
"fastq files : ${params.fastq}"
Channel
.
fromPath
(
params
.
fastq
)
.
ifEmpty
{
error
"Cannot find any fastq files matching: ${params.fastq}"
}
.
set
{
fastq_files
}
fastq_files
.
into
{
fastq_files_adaptor
;
fastq_files_fastq
}
process
fastqc_fastq
{
tag
"$reads.baseName"
publishDir
"results/fastq_SE/fastqc/raw"
,
mode:
'copy'
input:
file
reads
from
fastq_files_fastq
output:
file
"*.{zip,html}"
into
fastqc_repport
script:
"""
fastqc --quiet --threads ${task.cpus} --format fastq --outdir ./ ${reads}
"""
}
process
adaptor_removal
{
tag
"$reads.baseName"
input:
file
reads
from
fastq_files_adaptor
output:
file
"*_cut.fastq.gz"
into
fastq_files_cut
script:
"""
cutadapt -a TGGAATTCTCGGGTGCCAAGG -g CCTTGGCACCCGAGAATTCCA \
-o ${reads.baseName}_cut.fastq.gz \
${reads} > ${reads.baseName}_report.txt
"""
}
fastq_files_cut
.
into
{
fastq_files_cut_randombp
;
fastq_files_cut_fastq
}
process
fastqc_fastq_cutadapt
{
tag
"$reads.baseName"
publishDir
"results/fastq_SE/fastqc/adaptor_removal/"
,
mode:
'copy'
input:
file
(
reads
)
from
fastq_files_cut_fastq
output:
file
"*.{zip,html}"
into
cutadapt_fastqc_repport
script:
"""
fastqc --quiet --threads ${task.cpus} --format fastq --outdir ./ ${reads}
"""
}
process
random_bases_4_trimming
{
tag
"$reads.baseName"
publishDir
"results/fastq_SE/adaptor_removal/"
,
mode:
'copy'
input:
file
reads
from
fastq_files_cut_randombp
output:
file
"*_cut4.fastq.gz"
into
fastq_files_cut4
script:
"""
cutadapt -u 4 -u -4 \
-o ${reads.baseName}_cut4.fastq.gz \
${reads[0]} > ${reads.baseName}_report.txt
"""
}
fastq_files_cut4
.
into
{
fastq_files_trim
;
fastq_files_cut4_fastq
}
process
fastqc_fastq_randombp
{
tag
"$reads.baseName"
publishDir
"results/fastq_SE/fastqc/random_bases_4_trimming/"
,
mode:
'copy'
input:
file
reads
from
fastq_files_cut4_fastq
output:
file
"*.{zip,html}"
into
randombp_fastqc_repport
script:
"""
fastqc --quiet --threads ${task.cpus} --format fastq --outdir ./ ${reads}
"""
}
/*
* urqt :
* Imputs : fastq files
* Output : fastq files
*/
/* quality trimming */
/*
* for single-end data
*/
process
trimming
{
tag
"${reads}"
cpus
4
publishDir
"results/fastq_SE/trimming/"
,
mode:
'copy'
input:
file
reads
from
fastq_files_trim
output:
file
"*_trim.fastq.gz"
into
fastq_files_urqt
script:
"""
UrQt --t 20 --m ${task.cpus} --gz \
--in ${reads} \
--out ${reads.baseName}_trim.fastq.gz \
> ${reads.baseName}_trimming_report.txt
"""
}
fastq_files_urqt
.
into
{
fastq_files_CCA
;
fastq_files_urqt_fastq
}
process
fastqc_fastq_urqt
{
tag
"$reads.baseName"
publishDir
"results/fastq_SE/fastqc/urqt/"
,
mode:
'copy'
input:
file
reads
from
fastq_files_urqt_fastq
output:
file
"*.{zip,html}"
into
urqt_fastqc_repport
script:
"""
fastqc --quiet --threads ${task.cpus} --format fastq --outdir ./ ${reads}
"""
}
process
CCA_removal
{
tag
"$reads.baseName"
input:
file
reads
from
fastq_files_CCA
output:
file
"*_cut_CCA.fastq.gz"
into
fastq_files_cut_CCA
script:
"""
cutadapt -a CCA -g TGG \
-o ${reads.baseName}_cut_CCA.fastq.gz \
${reads} > ${reads.baseName}_report.txt
"""
}
/*
* Bowtie2 :
* Imputs : fastq files
* Imputs : fasta files
* Output : bam files
*/
/* fasta indexing */
params
.
fasta
=
"$baseDir/data/bam/*.fasta"
log
.
info
"fasta files : ${params.fasta}"
Channel
.
fromPath
(
params
.
fasta
)
.
ifEmpty
{
error
"Cannot find any bam files matching: ${params.fasta}"
}
.
set
{
fasta_file
}
process
index_fasta
{
tag
"$fasta.baseName"
cpus
4
publishDir
"results/mapping/index/"
,
mode:
'copy'
input:
file
fasta
from
fasta_file
output:
file
"*.index*"
into
index_files
script:
"""
bowtie2-build --threads ${task.cpus} ${fasta} ${fasta.baseName}.index &> ${fasta.baseName}_bowtie2_report.txt
if grep -q "Error" ${fasta.baseName}_bowtie2_report.txt; then
exit 1
fi
"""
}
/*
* for single-end data
*/
process
mapping_fastq
{
tag
"$reads.baseName"
cpus
4
publishDir
"results/mapping_SE/bams/"
,
mode:
'copy'
input:
file
reads
from
fastq_files_cut_CCA
file
index
from
index_files
.
collect
()
output:
file
"*.bam"
into
bam_files
file
"*_bowtie2_report.txt"
into
mapping_fastq_report
script:
index_id
=
index
[
0
]
for
(
index_file
in
index
)
{
if
(
index_file
=~
/.*\.1\.bt2/
&&
!(
index_file
=~
/.*\.rev\.1\.bt2/
)
)
{
index_id
=
(
index_file
=~
/(.*)\.1\.bt2/
)[
0
][
1
]
}
}
"""
bowtie2 --very-sensitive -p ${task.cpus} -x ${index_id} \
-U ${reads} 2> \
${reads.baseName}_bowtie2_report.txt | \
samtools view -Sb - > ${reads.baseName}.bam
if grep -q "Error" ${reads.baseName}_bowtie2_report.txt; then
exit 1
fi
"""
}
/*
* filtering mapping quality >= 2
*/
process
quality_filtered_bam
{
tag
"$bam.baseName"
cpus
2
input:
file
bam
from
bam_files
output:
file
"*_filtered.bam*"
into
filtered_bam_files
script:
"""
samtools view -hb -q 2 ${bam} > ${bam}_filtered.bam
"""
}
/* MultiQC */
process
multiqc
{
tag
"$repport"
publishDir
"results/fastq_SE/multiqc/"
,
mode:
'copy'
cpus
=
1
input:
file
repport
from
fastqc_repport
.
collect
()
file
repport_urqt
from
urqt_fastqc_repport
.
collect
()
file
repport_cutadapt
from
cutadapt_fastqc_repport
.
collect
()
file
repport_randombp
from
randombp_fastqc_repport
.
collect
()
output:
file
"*multiqc_*"
into
multiqc_report
script:
"""
multiqc -f .
"""
}
/* bams sorting */
process
sort_bam
{
tag
"$bam.baseName"
cpus
4
publishDir
"results/mapping_SE/bams/"
,
mode:
'copy'
input:
file
bam
from
filtered_bam_files
output:
file
"*_sorted.bam"
into
sorted_bam_files
script:
"""
samtools sort -@ ${task.cpus} -O BAM -o ${bam.baseName}_sorted.bam ${bam}
"""
}
/* bams indexing */
process
index_bam
{
tag
"$bam.baseName"
publishDir
"results/mapping_SE/bams/"
,
mode:
'copy'
input:
file
bam
from
sorted_bam_files
output:
file
"*bam*"
into
indexed_bam_file
script:
"""
samtools index ${bam}
"""
}
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