Rep (2) : a good PCR mimic?
sge1 at york.ac.uk
Fri Jun 28 09:19:29 EST 1996
Can I add that I have used several methods to generate internal standards
for competitive PCR and low stringency amplification of nontemplate was
the easiest and cheapest. However none of the literature on this that I
have seen tells you to check for both primer sites. With low stringency
PCR it is possible that some products have the same primer site at either
end. If the two primers have different Tm then it is likely that
proportional co-amplification will not occur. It is easy to check the
primer sites of the internal standard by trying to PCR it with the two
primers singly and in combination. Only fragments amplified in the
presence of both primers should be used.
University of York
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