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  • Lilach
    Member
    • Sep 2011
    • 20

    #1

    how to align concatenated and sheared sequences?

    Hi,

    I enriched DNA targets by PCR, them concatenated (ligated) the DNA and sheared it to get ~300 bp sequences.
    The problem is that each read may include sequences from two different genomic regions.
    The sequencing was performed with HiSeq PE x 100.
    How can I do the alignment with free software?

    In regular enrichments (RNA hybridization) I'm just using bwa and samtools, but I don't know if it is suitable for this case as well?

    Thanks,
    Lilach
  • swbarnes2
    Senior Member
    • May 2008
    • 910

    #2
    I think that's going to be ugly; with short reads, you need speed, and you can't break apart 100 Mreads a thousand ways each to see how the two ends might best map on separate reference sequences.

    You could try making concatenated reference sequence for those reads to align to.

    Comment

    • Lilach
      Member
      • Sep 2011
      • 20

      #3
      Thanks for the answer, although I hope there will be an easier way...
      I know what will be the PCR products. Is there any efficient algorithm that can concatenate these known sequences in all possible ways and "break" to pieces, to see what I get? Then I could align my results to this database?
      But I still have to "label" somehow each PCR product before the concatenation to know where it came from? or at least where are the end points or original sequences?

      Comment

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