Hi everyone,
We've been having problems with using Vent polymerase in a high GC
background. I study Pseudomonas aeruginosa (ca 69%GC), and routinely use 5%
DMSO in my PCR's, with Taq - without DMSO there are no bands (it's a bit of
a magic bullet effect!). When we shifted to Vent, we found that the enzyme
seems to be sensitive to DMSO, as we got nothing - and of course no bands
WITHOUT DMSO either! Does anyone have an easy way round this ( ways to
stabilize Vent in the presence of DMSO or other solvents, for instance)?
Thanks a lot,
Michael Kertesz
ETH-Microbiology,
ETH-Zentrum/LFV,
CH-8092 Zurich, Switzerland
tel: +41-1-632 3357
fax: +41-1-632 1148
e-mail: kertesz at micro.biol.ethz.ch
