HELP! VENT / Pfu amplification

C.Pical C.Pical at shef.ac.uk
Wed Jun 26 05:03:22 EST 1996


Yes, I have had this problem, using Pfu. I managed to get some 
amplification, by using tha reaction buffer provided with Taq, and 
doubling the extension time (Pfu has proof-reading activity). Still, 
the yield was poor compared to Taq (about 20% I would say). But that 
was enough for successful cloning. I have not tried ro increase the 
yield, by playing with MgCl2,... because Pfu is expensive. Still, I 
think there must be a way to get good yields. Any other suggestions 
anyone?




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