Uploaded image for project: 'UGENE'
  1. UGENE
  2. UGENE-771

Incorrect fragment digestion with non-palindromic restriction sites

    XMLWordPrintable

    Details

    • Type: Bug
    • Status: Closed
    • Priority: Major
    • Resolution: Fixed
    • Affects Version/s: None
    • Fix Version/s: 1.10.3
    • Component/s: Basic-Nucl
    • Labels:
    • Tests Type:
      Functional/Unit
    • Sprint:
      TEST-2013/08/15, TEST-2013/08/29
    • Affect Type:
      Userdefined

      Description

      Overhang's strand is not detected properly when the cut occurs on the negative strand of non-palindromic site.

      From user's email:

      "I've run into an issue with a non-palindromic restriction enzyme,
      BsaI, when using the "Digest into Fragments" function. The cutting
      pattern is below:
      http://www.neb.com/nebecomm/products/productR0535.asp
      When the recognition sequence GGTCTC is on the primary strand, cutting
      goes correctly:
      GGTCTCTAATGC
      CCAGAGATTACG
      becomes
      GGTCTCAXXXX
      CCAGAGATTAC
      (where X means absent bases)

      However! if the recognition sequence is not on the primary strand, e.g.
      AGCGCTGAGACCCGATAT
      TCGCGACTCTGGGCTATA
      the cut should be
      XGCGCTGAGACCCGATAT
      XXXXX ACTCTGGGCTATA
      (to the 3' end of the recognition sequence on the reverse
      complementary strand) but the result given by the "digest into
      fragments function" :
      AGCGCTGAGACCCXXXXX
      TCGCGACTCTGGGCTATX
      (which is to the 5' end of the recognition sequence on the reverse
      complementary strand.)
      In other words, despite the fact that GGTCTC is on the rev. compl.
      strand, it is correctly locating the sequence... and then cutting as
      if it's on the primary strand. Looking at other randomly chosen
      non-palindromic enzymes like BceAI, this problem does not seem to be
      limited to BsaI. Anyway, this is obviously a problem if you are
      trying to use the cloning functions and you are using a
      non-palindromic enzyme. (I don't seem to be having this problem with
      any of the palindromic enzymes I've used recently - NcoI, EcoRI,
      SacII.)""

        Attachments

          Activity

            People

            Assignee:
            kokonech Konstantin Okonechnikov
            Reporter:
            kokonech Konstantin Okonechnikov
            Assigned Tester:
            Artyom Savchenko
            Watchers:
            2 Start watching this issue

              Dates

              Created:
              Updated:
              Resolved: