Rep (2) : a good PCR mimic?

SG Edwards 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.

Simon Edwards
University of York
UK



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