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cuffdiff2 fails to find diff expressed genes!!??

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  • cuffdiff2 fails to find diff expressed genes!!??

    Hello,

    I processed 8 breast cancer samples, 4ER+ and 4ER- using tophat2/cufflinks2/cuffmerge2/cuffdiff2/cummeRbund.

    The expression of the gene GREB1 is nearly equal to 0 for the 4 ER-
    and ~100 for ER+:

    cuff <- readCufflinks()
    expr=repFpkmMatrix(genes(cuff))
    expr[rownames(expr)=='XLOC_011466',]
    Neg_0 Neg_1 Neg_2 Neg_3 Pos_0 Pos_1 Pos_2 Pos_3
    XLOC_011466 0.182529 0.791468 0.721075 0.15861 35.9609 231.247 64.1414 39.4893

    However, the result of cuffdiff is:
    diffData(getGene(cuff, 'GREB1'))
    gene_id sample_1 sample_2 status value_1 value_2
    log2_fold_change test_stat p_value q_value significant
    1 XLOC_011466 Neg Pos OK 0.460534 92.726
    7.65352 -1.1267 0.25987 0.999954 no

    How can you explain this. It seems not consistent for me. I would have
    expected a nice q value, a significant test, etc.

    thanks for feedbacks,
    colin

    ps. data were downloaded from SRA/GEO : GSE27003 and are available at inSilico: https://insilico.ulb.ac.be/app/browse?q=GSE27003

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