T7 DNA pol for SD mutagenesis

[Karl Fischer]

In article <chauser-1412941628200001 at bglab.botany.duke.edu>,
chauser at acpub.duke.edu (Chuck Hauser) wrote:

> I have heard that some supplier's T7 DNA polymerase will not work in
> Kunkle mutagenesis.  I know that the 3' exo is necessary, and thus
> sequenase does not work, but rumor is that some suppliers holoenzyme is
> inefficient at repairing.  Does anyone have experience with one which they
> know will work?
>                   thanks, chuck

We don't use no fargin' T7 Dpol <insert evil grin here>. 

Seriously....I use T4 Dpol/T4 ligase in a single tube extension on a
tandem oligo mutagenesis system. Works great (less filling...sorry -
couldn't resist).  Drop a line if you want the procedure.

Cheers and beers

Karl

-- 
Karl Fischer
kfischer at gpu.srv.ualberta.ca
tyr-2 at bones.biochem.ualberta.ca