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  • tujchl
    Member
    • Sep 2009
    • 74

    #1

    strange output from RSEM

    hi,
    I use RSEM (http://deweylab.biostat.wisc.edu/rsem/README.html) to analysis my RNA-seq data and want to summarize expression level of each RefSeq ID.
    the command I used is as below:
    rsem-calculate-expression -p 7 --fragment-length-min 51 --fragment-length-max 51 --fragment-length-mean 51 --fragment-length-sd 0
    BTW, the length of all my reads is 51bp

    I found that many isoform from the same gene got quite different TPM and FPKM. An example is as below:
    transcript_id gene_id length effective_length expected_count TPM FPKM IsoPct
    NM_001130147.1 CCDC77 2359 2309.00 317.27 16.83 11.21 100.00
    NM_001130148.1 CCDC77 2305 2255.00 2.23 0.12 0.08 100.00

    as you see. NM_001130147.1 and NM_001130148.1 are different isoform of CCDC77 and the mRNA seq is quite similar with each other. but the TPM as well as FPKM is quite different which I regard it as err.

    Does anyone meet the err? can you help me?
  • dpryan
    Devon Ryan
    • Jul 2011
    • 3478

    #2
    Why do you consider that to be an error? Typically there's only one major isoform.

    Comment

    • tujchl
      Member
      • Sep 2009
      • 74

      #3
      thank you for your replying, dpryan:

      because the mRNA seq of these two isoforms are nearly the same. Most of the reads mapped to NM_001130147.1 can also be mapped to NM_001130148.1. So I think the expression level of these two isoforms should be nearly the same.

      Am I right?

      Comment

      • dpryan
        Devon Ryan
        • Jul 2011
        • 3478

        #4
        No, just because most of the reads can map to both doesn't mean the expression was more or less the same. That's the entire point of programs like RSEM, have a read through the paper for details.

        Comment

        • tujchl
          Member
          • Sep 2009
          • 74

          #5
          Thank you, dpryan
          To be honest, I didn`t read the papper. Now I think it`s the time to read it.

          Comment

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