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  • szaks
    Junior Member
    • Mar 2013
    • 1

    Cufflinks - LOWDATA and NOTEST in *.diff files

    After running cuffdiff, in all of the *.diff files the test status is “LOWDATA”, “NOTEST”, or, to a smaller extent, “FAIL.” On the other hand, the *_exp.diff files are fine; many of the test status are "OK". Any thoughts why this might be happening?

    The LOWDATA status is strange, as all of my targeted genes have >1000x coverage. Of note, this is targeted RNASeq data set; we only sequenced 30 genes. Maybe the targeted nature of the data set is the problem? The .gtf file that I'm using has all human genes in it; should I parse out our targeted genes of interest and use the smaller gtf file?

    I'm using cufflinks v2.0.2.

    Thanks in advance for any advice, guidance, or direction!
  • ramiro2k
    Member
    • Mar 2012
    • 12

    #2
    Any answers?

    Did you find a solution to this?? I am experiencing the same problem...

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    • GATTACAT
      Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
      by GATTACAT
      Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
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    • SEQadmin2
      Nine Things a Sample Prep Scientist Thinks About Before Sequencing
      by SEQadmin2


      I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.

      Here are nine questions we think about, in roughly the order they matter, before...
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