Increasing PCR specificity

[Leslie A. Johnston-Dow]

Michael and all,

A nice discussion of the specificty of PCR reactions can be found in the 
October 1991, Issue 7 of the PE-Cetus publication 'Amplifications' and a 
couple of nice, indepth reviews (W. Bloch,'Biochemistry'1991, 30(11)
pp2735-2747 and H.Erlich et al 'Science'1991 252 pp1643-1651). The reviews
focus on the specifics of the PCR process.

I have tried several approaches to increase PCR specificity when amplifying  
from human genomic DNA and have found the best approach for my work is to 
increase the annealing temperature 5degC in the first 5 cycles. This was 
described by Gualberto Ruano in the PE-Cetus article. This provides very 
clean PCR products, which I then sequence on our fluorescent sequencer.

Good Luck!
Leslie  


Applied Biosystems, Inc
(415) 570-6667
ldow at apldbio.com