Request: Vent PCR: advice on optimizing Longish PCR

ladasky at my-dejanews.com ladasky at my-dejanews.com
Mon Mar 15 19:25:23 EST 1999


In article <7cjsgn$lq4$1 at nntp4.u.washington.edu>,
  polliwog at u.washington.edu (S. Findley) wrote:
>
> I am trying to amplify fragments of 2-3 kB with NEB's Vent polymerase.
> I have heard that this enzyme doesn't like to please researchers trying to
> amplify longer fragments.
>
> What conditions have proven successful in optimizing/extending Vent
> performance?


Mixing with Taq polymerase, for one!

The long PCR protocols typically use between 1% and 10% Taq polymerase in
conjunction with Vent (NEB) or Pwo (Boehringer) or Pfu (Stratagene).  This
way you get the error correction of the high-fidelity enzym and the proces-
sivity of Taq.

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