Vent PCR Woes

Tony Hodge tph at mrc-lmb.cam.ac.uk
Wed Jun 15 05:22:12 EST 1994


Subject: Re: Vent PCR Woes
From: Hong Dang, hdang at cns.neusc.bcm.tmc.edu
Date: 13 Jun 1994 15:36:37 GMT
In article <2thue5$nct at gazette.bcm.tmc.edu> Hong Dang,
hdang at cns.neusc.bcm.tmc.edu writes:
>Dave,
>You didn't do anything wrong, to my experience. I tried Vent and
DeepVent, never
>got any decent PCR. Stratagene's Pfu worked a little better, but
nothing to really
>depend on. One thing you can try if you have to have them work, is
10% glycerol.
>Apparently, 3'-5' exo proofreading activity makes them very
"picky", so there is
>one more enzyme you can try, Ultma (Perkin-Elmar) has a reduced exo
activity which
>allows it to be more efficient, so I heard, but it is just like
mixing Taq and an
>axo enzyme (Barnes' paper).

Hong,

??????

In a number of head to head Taq vs Vent experiments I nearly always
got more, higher quality (by cloning and sequencing) product from
the Vent.  The only time I had problems was in trying to PCR Screen
with Vent - when it worrks it gave masses of product but on
occaision failed completely so I now only use Taq for this
application.

I'm sold on it anyway!

Tony.


.
Tony P Hodge
Structural Studies Division
Medical Research Council Laboratory of Molecular Biology
Hills Road
Cambridge
CB2  2QH
UK

Phone (0223)  402260

Fax     (0223)  213556



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