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  • Big_Cheese
    Junior Member
    • Jul 2012
    • 7

    #1

    Isoforms in Cufflinks

    Hi,

    I am using cuffdiff and Deseq/edgeR to compare Gene expression levels between mice kidneys between a control and an affected group.

    Both outputs are comparable, the difference is that cufflinks also finds gene isoforms.
    So for example if you compare a random gene you will probably get a 1:1 ratio with the raw read data. But if you look it up in cuffdiff, there are sometimes up to 5 isoforms with very expression ratios. But if you add them together you will get a 1:1 ratio again.
    There are some cases where the isoform seems to "switch".
    So in the control group you have average 5 fpkm for isoform A and 0 for isoform B,
    but in the affected group it's the other way around: isoform B has average 5 fpkm and isoform A has nothing.


    Most of the times when the isoform varies, its with very low FPKM
    example:
    From Gene X,
    isoform A has 40 FPKM in control and 38 in affected
    isoform B has 0 FPKM in control and 0.5 in affected
    isoform C has 5 in control and 2 in affected

    if you add these values, you get the same ratio for Gene X you would get in the raw reads file.

    So my question now is if that means something. Can different isoforms affect the kidneys, or is what I see nothing out of the ordinary and the variations are natural/statistical Errors

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