In article <48s9d4$ikg at ipgate.le.ac.uk>, mark leyland <ML27 at le.ac.uk> wrote:
> I've also had this problem in the past and have found that it is often
best to try using the native (non-recombinant) version of Pfu=
> DNA polymerase. For some reason the native version of the enzyme will
often amplify DNA that the recombinant version won't.
I concur. My right and honourable friend expressed feelings of thousands
oppressed users of cloned Pfu polymerase. For reasons that remain obscure
cloned Pfu appears to suck. A lot. There are other choices of high
fidelity enzymes for PCR - we used to rely heavily on Tth polymerase, now
most of the work is done using Pwo pol. Both give good yields, very
reproducible and we now have sequenced more than 12 kb of Pwo products to
attest to its high fidelity.
The lord of the yeast