Convert modENCODE PET RNA-seq SAM to pileup bigWig

convertModENCODERnaSeqToBW.sh

#!/bin/bash

if [ $# -lt 1 ]; then
    echo `basename $0` "<PET RNAseq SAM filename>"
    exit
fi

F=$1

# chromosome length file
G=ce10.len

# a temporary name
TNAME=`mktemp -u`
TNAME=$1`basename $TNAME`

# convert SAM to BAM
samtools view -bS $F -o ${TNAME}.bam
# sort BAM
samtools sort ${TNAME}.bam ${TNAME}.sorted
# keep only paired reads
samtools view -b -f 2 ${TNAME}.sorted.bam > ${TNAME}.sorted.paired.bam
# convert to bed12
bamToBed -i ${TNAME}.sorted.paired.bam -bed12 > ${TNAME}.sorted.paired.bed12
# fix chromosome names from wormbase/ensembl convention to UCSC style
perl -pi -e '$_="chr".$_;s/MtDNA/M/' ${TNAME}.sorted.paired.bed12

# count total reads number and calculate scaling factor for RPM values
TOTAL=`wc -l ${TNAME}.sorted.paired.bed12|cut -f 1 -d " "`
SCALE_FACTOR=`echo "scale=5;1000000.0/"$TOTAL | bc`

# compute coverage file in bedGraph, for RPM values
genomeCoverageBed -split -scale ${SCALE_FACTOR} -i ${TNAME}.sorted.paired.bed12 -bga -g ${G} > ${F}.bdg

# convert bedGraph to bigWig 
bedClip ${F}.bdg ${G} ${TNAME}.bdg.clip
bedGraphToBigWig ${TNAME}.bdg.clip ${G} ${F}.bw

# clean temporary files
rm -f ${TNAME}.*

echo "Check ${F}.bw"