Team:University of Ottawa/12 June 2008

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Today in the lab

Dan and Matt

Results from the gel
  • Showed that PCR did not work
  • Source of problems can be from:
    1. The sequence that we have on hand is not the plasmid that we have
    To resolve this we will digest the plasmid with HindIII
    2. The PCR is not working
    We will run a PCR temperature gradient.
  • If the second test is inconclusive we will order new primers.

    • Digestion of 7/8 plasmids using HindIII'

  • Worked out as expected, there is a high probability that we have the correct plasmid

    • PCR gradient between 55 and 70 degrees C

  • Gel showed that there are very faint expected PCR products between ~56-60 degrees C
  • We will narrow our gradient and try the PCR optimization again, although we think that it is unlikely that there is a problem with the PCR because we are using the enhanced phusion enzyme.
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