1 Commits

2 changed files with 53 additions and 18 deletions

View File

@ -0,0 +1,53 @@
#!/bin/bash
#
# Align reads against reference genome.
STORAGE="/groups/umcg-griac/tmp04/rawdata/$(whoami)/step3"
# Store genome index in this location:.
GENOME_INDEX="${STORAGE}/genome_index"
mkdir -p "${GENOME_INDEX}"
# Store the generated `Aligned.sortedByCoord.out.bam` in this dir.
ALIGNMENT_OUTPUT="${STORAGE}/alignment"
mkdir -p "${ALIGNMENT_OUTPUT}"
# 1) Generate genome index.
#
# N.B.:
# - We're assuming a read size of 100 bp (--sjdbOverhang 100). Refer back to the
# previous quality control steps if you are unsure about the size. In case of
# reads of varying length, the ideal value is max(ReadLength)-1.
# - We're using gzip compressed reference data (--readFilesCommand zcat), i.e.,
# .gtf.gz and fa.gz. If not, you can remove the `zcat` flag.
# Storage location reference data (in this case on calculon).
REFERENCE_DATA="/groups/umcg-griac/prm02/rawdata/reference/genome"
GTF_FILE="${REFERENCE_DATA}/Homo_sapiens.GRCh38.100.gtf.gz"
FASTA_FILE="${REFERENCE_DATA}/Homo_sapiens.GRCh38.dna.primary_assembly.fa.gz"
STAR \
--runThreadN 8 \
--runMode genomeGenerate \
--readFilesCommand zcat \
--sjdbOverhang 100 \
--genomeFastaFiles ${FASTA_FILE} \
--sjdbGTFfile ${GTF_FILE} \
--genomeDir ${GENOME_INDEX}
# 2) Do the actual alignment.
#
# N.B.:
# - We are assuming paired-end, gzip compressed (--readFilesCommand zcat) FastQ
# files.
# THe compressed paired-end FastQ's that we are aligning.
R1="sample1_R1.fastq.gz"
R2="sample1_R2.fastq.gz"
STAR \
--runThreadN 8 \
--readFilesCommand zcat \
--readFilesIn "${R1}" "${R2}" \
--outSAMtype BAM SortedByCoordinate \
--genomeDir ${GENOME_INDEX} \
--outFileNamePrefix "${ALIGNMENT_OUTPUT}"

View File

@ -1,18 +0,0 @@
## Generate Raw Counts using FeatureCounts/HTSeqCounts
## Load package Subread (packge within which FeatureCounts is present)
+ ml Subread/1.6.4-foss-2015b FeatureCounts
## Features used in FeatureCounts :
## -s : reversely stranded
## -T : number of threads used
## -a : reference genome
## -o : output file
+ featureCounts \
+ --a "path to reference genome" \
+ --T 6 \
+ --o "path to output file" \
+ --s 2