Thanks for the reply…we'll discuss your suggestions and see what works!
On Sunday, May 26, 2013 1:10:27 PM UTC-4, cory....@gmail.com wrote:
Assuming you added all of the correct ingredients and you know your--
primers work, I have two suggestions
1) Make sure your pipettes are calibrated and that you pipette
accurately. When I run bio classes for beginners this seems to be one
of the biggest hurdles to getting PCRs to work. Taq polymerase is
very sensitive to pH, salt concentration, etc so pipetting accurately
is critical.
2) Unless you know the concentration of your template DNA, make a
couple dilutions of your template. Start with a 100x dilution and see
if that works. FYI, high concentration of DNA can inhibit PCR or
cause non-specific amplification.
-cory
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