pwo polymaerase

Kirsty Harvey harvey at ch1.fgg.eur.nl
Fri Aug 18 09:26:12 EST 2000


Hi all,

I have tried to use PWO (proofreading) DNA polymerase for PCR cloning.
With 'normal' Taq polymerase I get a nice one band PCR product. When I
use PWO with similar PCR conditions I get a smear on my agarose gel. I
have tried increasing the Mg-concentration, increasing the annealing
temp., changing both the enzyme and template concentration, the number
of cycles and elongation time, with and without DMSO. So probably most
common problems have been eliminated. Has anyone any other ideas???? I
am out of any...

Claudia

--
K N Harvey
Dept of Cell Biology & Genetics
PO Box 1738
3000DR Rotterdam
The Netherlands
Tel: + 31 10 4087282
Fax: + 31 10 4087282

mailto harvey at ch1.fgg.eur.nl







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