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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