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RPKM normalization - Perl
By EagleEye 3077 days ago Comments (2)
#!/usr/bin/perl
#use strict;
#use warnings;
#Author: Santhilal Subhash
#Contact: santhilal.subhash@gu.se
#RPKM for RNAseq V1.3
#USAGE for sample input provided: perl rpkm_script_beta.pl sample_count_test.count 2:9 15 > sample_count_test.rpkm
#USAGE: perl rpkm_script_beta.pl input_count_file.txt ActualColumnStart:ActualColumnEnd ColumnGeneLength > results.rpkm
open $fh1, '<', $ARGV[0] or die $!;
open $fh2, '<', $ARGV[0] or die $!;
$total = 0;
$count = 0;
$cols=$ARGV[1];
$len_col=$ARGV[2];
while (<$fh1>)
{
@array=split("\t");
($cstart,$cend)=split(":",$cols);
for($i=$cstart-1;$i<=$cend-1;$i++)
{
$libarray[$i] += $array[$i];
}
}
while (<$fh2>)
{
$next = <>;
if ($next =~ /^#/)
{
$header=$next;
$header =~ s/#//;
print "$header";
}
if ($next !~ /^#/)
{
@array=split("\t");
($cstart,$cend)=split(":",$cols);
for($i=$cstart-1;$i<=$cend-1;$i++)
{
if($array[$i]!=0)
{
$array_rpkm[$i]=((1000000000*$array[$i])/($libarray[$i]*$array[$len_col-1]));
}
else
{
$array_rpkm[$i]=0;
}
}
local $" = "\t";
print "$array[$cstart-2]@array_rpkm\t@array[$cend..$#array]";
}
}
Comments
Usage: perl rpkm_script_beta.pl sample_count_test.count 2:9 28 > sample_count_test.rpkm
Colum1 in the input matrix is gene name, column 2 to 9 samples with corresponding counts and column 28 is length of each corresponding gene
More info: https://github.com/santhilalsubhash/rpkm_rnaseq_count
Sample input:
https://github.com/santhilalsubhash/rpkm_rnaseq_count/blob/master/sample_count_test.count
Using R script,
http://bioinformaticsonline.com/snippets/view/28042/rpkm-normalization-r-script