Optimal Conditions for Boehringer PWO enzyme in PCR

Jonathan Patrick Burke 075jon at chiron.wits.ac.za
Tue Oct 18 06:47:49 EST 1994


I am interested in using the Boehringer Pwo polymerase enzyme in my PCR 
reactions to amplify a 350bp fragment from whole genomic human DNA. I have 
tried a reaction using a setup analogous to the normal and successfulTaq 
polymerase reaction. The Taq works but the Pwo does not. The blurb from 
Boehringer states that the reaction conditions need to be optimised. The 
number of possible variations is very large; does anyone have any 
experience with this enzyme that would give me a starting point? The other 
claim is that the Pwo amplicon is blunt ended and therefore easier to 
subclone. Is this the case?
Thanking you in advance for any help

Jonathan Burke

Univ Witwatersrand Medical School
Johannesburg
South Africa



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