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.
-----------== Posted via Deja News, The Discussion Network ==----------
http://www.dejanews.com/ Search, Read, Discuss, or Start Your Own
More information about the Methods
mailing list
Send comments to us at biosci-help [At] net.bio.net