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  • Merged illumina data

    I encountered something rather odd and I am not certain that this is an illumina artefact or that it is true what I see.

    I was curious if it is possible that when working with RNA sequences, is it possible that two separate RNA strands are merged against each other?

    That for example when sequence:

    ATGCATGCATGCATGCATGCATGC

    and

    GATCGATCGATCGATCGATCGATC

    become

    [Adapter]ATGCATGCATGCATGCATGCATGCGATCGATCGATCGATCGATCGATC[adapter]

  • #2
    How were these RNA libraries prepared? If the ligation method was used (directional RNA-Seq instead of TruSeq) to add the sequencing adapters, it is possible for two inserts to ligate together during the adapter ligation reaction.

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    • #3
      I will look up the exact procedure but what I found out so far is that of the 5.700.000 reads 290.000 of them had this odd result. Which is 5% of the total amount of reads. Would this be a lot if this was an illumina artefact?

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      • #4
        How did you find those reads which originated from two different strands at the first place?

        Wei

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