Taq - not so promiscuous???

rpgrant at vax.path.ox.ac.uk rpgrant at vax.path.ox.ac.uk
Mon Dec 14 08:33:12 EST 1992


Following the recent discussion about various polymerases used in PCR, I'd 
like to add my tuppence.
Recently I amplified, cloned and sequenced two lengths of DNA, 1.35kb and 
1.9kb, using Taq.   To my surprise, I found that the sequence of the first 
clone in both experiments was 100.000% correct.  (BTW, the amplifications also 
incorporated restriction sites and eukaryotic initiation sequences).

Now, was I exceptionally lucky, or was it the little trick my boss suggested? 
Namely, I used 50uM dNTP in the amplification reactions instead of the normal 
200uM.  
It would be interesting to see if this modification increases fidelity in 
general......
BFN,
Richard 



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