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  • NicoBxl
    not just another member
    • Aug 2010
    • 264

    #1

    Illumina stranded RNA-Seq background problem

    Hi everybody,

    We perform RNA-Seq experiments using illumina TruSeq total RNA stranded (with RiboZero) and we have strange results compared to previous samples. We sequenced on a HiSeq2000 2x100bp.

    Here a picture of the alignment results :



    The two first lane are a previously sequenced sample (same kit) and the two below are a new one. You can see that on the new ones, there is a background. And it is almost on the whole genome like that...

    Someone has an idea of ​​where the problem comes from?

    Thanks a lot,

    N.
  • Bukowski
    Senior Member
    • Jan 2010
    • 388

    #2
    Well if it's all over the genome, I'd suggest it was genomic DNA. Contamination or sample mix-up.

    Comment

    • Genohub
      Registered Vendor
      • Mar 2013
      • 210

      #3
      Looks like genomic DNA contamination. Did you treat your RNA with DNAse?

      Comment

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