Site-directed mutagenesis - with Taq? (Jul/30/2006 )

Maybe this will be a stupid question but is it possible to use Taq instead of Pfu? I know thath Taq mutation frequency is higher but I have sequenced hundreds of amplicons produced by Taq and I did not see additional mutations. Then how high is the possibility that it will produce mistakes in my 1,3kB long insert?

You can't do the Quikchange protocol with Taq. Quikchange relies on the enzyme stopping when it encounters the bound 5' end of the primer after copying the entire vector. Taq will displace the 5' primer end and keep going. Use Pfu, Phusion, or other enzymes which do not have displacement activity.

thanks... everyday i can learn something new