Hello
I have 400 fastq files from different samples in two sequencing runs. Both runs were on Illumina Hiseq. How i can merge the .fastq files of both runs for each sample, and in one step..For sure we have to keep R1 and R2 separate ..I know that we can just merge the .fastq files of both runs using cat..but i have to use this only for one sample…and than i have to repeat this many times for all the samples..and i have more than 100 samples... What command do i use ?? any folder to prepare?
Thank you for helping me
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written
by
rim.klabi •
1 answer
This is a little perl script I use. It works off the assumption that everything is in the same folder, and everything before the 'S\d+' is the name. It will cat together everything with the same name
foreach my $sample (@dir) {
next unless $sample =~ /.gz/;
my ($shortname) = $sample =~ /(\S+)_S\d+_L\d+_R\d_\d\d\d.fastq.gz/;
$hash{$shortname}++;
}
foreach my $key(keys(%hash)) {
mkdir $key;
my $temp = $dir . "/" . "$key" . "*.gz";
system("cd $key; cat $temp | STAR --genomeDir $genomeDir --sjdbGTFfile $gtf --readFilesIn - --readFilesCommand zcat --quantMode TranscriptomeSAM GeneCounts --outSAMunmapped Within --outSAMtype BAM SortedByCoordinate -- runThreadN 20 --limitBAMsortRAM 1001279989; cd ..;");
}
Thank you i will try ..
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You'll need a for loop for this, and to figure out how to write this command we need to know how your files are named, which naming pattern you use to distinguish the samples/lanes/read direction.
Why are you merging reads again?
OP is not merging the reads but merging file pieces for a sample.
bcl2fastqused to break files up in 2 million read chunks in past.maybe im just being too literal here, but he seems to be saying that he's merging reads from two independent runs. and if thats the case, then he probably should be treating each run as a technical replicate, or consider merging after correcting for batch effects.
I have to merge reads from two runs in order to increase the reads number ..
1, prepare two folders, and the same sample in two runs should share the same file name. And create an
combinedfolder.2, loop the folder of first run, for each file name you get, do 3
3, do
cat run1/samplename.R1.fq run2/samplename.R1.fq > combined/samplename.R1.fqandcat run1/samplename.R2.fq run2/samplename.R2.fq > combined/samplename.R2.fq