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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