I can actually align to the part of transcriptome that I want (first I have to make fasta file and use it as reference file to align to). I can also make gtf file using the same file and count using that. but in both cases it returns 0 counts. so I thought maybe I should align to the genome and count on the reads that mapped to that part. but here I do not know how to do that.
I have RNAseq data and trying to count the reads that mapped only to a part of 5'UTR (in fact the whole 5'UTR except the first 50 nts). do you know how to do that?
2 answers
Hi Sara,
Did you align the raw reads to a reference genome? if so what is the current format of your aligned reads?
Tarek
Hello Sara,
once you have mapped the data against the genome of your interest, you can simply use FeatureCounts program for instance:
featureCounts -a yourGTF.gtf -t UTR5 -g transcript_id -o counts_UTR5_transcripts.txt
The -t option allows you to select a type of features to be counted. The -g option allows you to choose "how to group your data". Using transcript_id (or something equivalent in your GTF/GFF), you tell to FeatureCounts: "ouput the counts for each transcript".
Also if you prefer to use R, you can use "Rsubread" package to count your reads using featureCounts () as mention by glihm.
T
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