HELP! VENT / Pfu amplification

Andrew Schrader andrews at
Wed Jun 26 18:53:19 EST 1996

Although some optimization was necessary, the yield of DNA (0.14 
to 2.3 kb frags) from amplification using Pfu is now comparable to 
that obtained with taq. I reduced my Mg++ by 2/3, added 1/10 vol of 
DMSO and reduced the dNTPs to 1/5 of that in a reaction using taq. 

I have also reduced the cycle number from 30 to 25 and raised the 
annealing temperature in most cases to 62C.

Hope this advice helps


Dr Andrew Schrader                                             
Molecular Biology and Hypertension Laboratory   
Department of Physiology (F13)
The University of Sydney, NSW, 2006
Phone 61 2 351 4599
Fax     61 2 351 2058
andrews at


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