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  • yueluo
    Member
    • Aug 2013
    • 82

    #1

    Wierd issue with Cufflinks 2.2.0

    Hi all,
    I'm having issues with results from cuffnorm.

    I had 4samples(R1-R4) with 3replicates each, so that's 12 PE fastqs
    R1= A1 A2 A3;
    R2= B1 B2 B3;
    R3= C1 C2 C3;
    R4= D1 D2 D3

    These were the steps I ran:
    1. Tophat / Cufflinks for each fq.(get 12 BAMS + 12 transcript.gtf)
    2. Cuffmerge to create merged.gtf.
    3. Cuffquant to create cxb.(each BAM vs merged.gtf, get 12 abundance.cxb)
    4. Cuffnorm with cxb and sample_sheet.
    And got (TableA1)
    I repeated step4, this time I treated replicates as individual samples.
    And got (TableA2)

    So now it seems cuffnorm re-arranged some of the columns, but FPKM values are the same.
    Since this run was performed on a cluster where I had no root-rights, I used a pre-compiled version of cufflinks-2.2.0.

    I then moved to our local server(with root-rights) and built cufflinks-2.2.0 from source.
    I used the same reference/cxb/merged.gtf files to rerun cuffnorm with the same parameters above. And got (TableB1) and (TableB2)

    To simplify things, all the tables where produced using '$> head genes.fpkm_table'. I also uploaded the "samples.table" of each run.
    Seems to me that Tables A2 and B2 agree with each other, but obviously something's wrong with Tables A1 and B2.

    Another thing I noticed : The pre-compiled version goes with SVN revision 4223, but the one I built from source is SVN revision 4222.

    I wonder if anyone else has run into this problem?
    Attached Files
    Last edited by yueluo; 03-31-2014, 07:46 PM. Reason: Missed attachments
  • yueluo
    Member
    • Aug 2013
    • 82

    #2
    I've been having problems browsing Cufflinks' Google Group. But my friend helped post it:


    Hope this doesn't cause any convenience.

    Comment

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