proofreading thermostable polymerases: Pfu vs. Pwo

[Paul W. Diaz]

I had to make around 50 "linker-scanning" mutagenized promoter 
constructs involving PCR amplication of a 800 bp promoter fragment.  
Only a couple of times did I detect infidelity due to Taq.  Curious 
about proofreading thermo-pols as you are, I tried Pfu and found that my 
PCR reactions would work only after substantial troubleshooting (and 
even then, not very well), so I returned to using Taq.  You might want 
to give Taq a try, unless the newer generations of proofreading pols 
work better.

John Rooney

UCB--Dept MCB