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  • RPKM and read counts from RNA-seq

    Hi All,
    I have alignment files from RNA-seq data in .sam format. I want to calculate read counts and RPKM values using RefSeq reference data (refseq.gft) file for several samples. I tried to use HTSeq, TopHat etc but I have not be able to get RPKM and read counts. Can you please suggest appropriate bioinformatics tools?
    Thanks for your help,
    Rakesh

  • #2
    Tophat is meant to align reads, not give you expression values.
    Try Cufflinks or rsem.
    HTSeq will give you the count data which you could convert into a normalized value if you wish.

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