HELP! VENT / Pfu amplification

[Andrew Schrader]

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

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

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