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ericmjl / protein-systematic-characterization

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Troubleshooting the PCR #10

Closed ericmjl closed 7 years ago

ericmjl commented 7 years ago

@Starnite - just thought of something.

Checked page 10 of the Agilent enzyme manual. http://www.agilent.com/cs/library/usermanuals/public/200550.pdf

There's two things we could try to optimize the PCR reaction that we haven't tried.

  1. Add DMSO. In molbio folklore, often this is a "magic" reagent that helps straighten out DNA. Mechanistically not sure how it works, but I know empirically that it can help.
  2. Try 40 cycles of PCR. That would equal to 210-fold amplification over 30 cycles.

What do you think?