re sequence polymorphisms

goldman%bchem.dnet at DXI.NIH.GOV goldman%bchem.dnet at DXI.NIH.GOV
Thu Nov 3 09:49:31 EST 1994


One issue to bare in mind when sequencing PCR products is fidelity of the 
the PCR reaction. Standard Taq is error prone, it has no editing function.
Therefore an early error will create a poylmorphism in the finl products. 
Pfu and Vent have exonuclease activity which removes errors. The latter two 
enzymes a much harder to use but supplimenting 1U of Taq with 1/200-1/100 U 
of Pfu is meant is be sufficient to reduce the errors significantly 
(see Barnes PNAS 91:2216 - beware in this paper the confusing mix of units 
and micro litres. Barnes used Taq at 25-35 U pel ul, but Pfu at 1U per ul).

So - if you want to sequence of PCR don't use Taq alone - maybe Xs clients 
are doing just that??/

ASHG



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