proofreading thermostable polymerases: Pfu vs. Pwo

Paul W. Diaz Pendragn at uclink2.berkeley.edu
Thu Nov 9 00:21:42 EST 1995


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





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