protocol colony PCR?

Rene Haanstra r.haanstra
Thu Sep 9 04:21:33 EST 1999


Dear Netters,

Does anyone have a protocol for colony PCR that works for most
bacteria? I experience a lot of problems when i try to do a 16S PCR
on colonies of bacteria. Two out of five colonies don't produce a product.
It seems to be not a problem in breaking the cells, since we get a product
with the same colonies when we perform a REP-PCR. In REPs the product
does not serve as a template, so the cells are broken. We think that the
DMSO in the REPs helps accessing the DNA, but we don't like to use DMSO
because it also has an effect on the annealing temperature and we don't
like to optimize the annealing temperature again.

Please help me out!

Rene Haanstra

´¯`·.¸¸.·´¯`·.¸¸.·´¯`·.¸¸.·´¯`·.¸¸.·´¯`·.¸¸.·´¯`·.¸¸.·´¯`·.¸¸.·´¯`·.¸¸.·´¯`·
.¸¸
Rene Haanstra
Center for Ecological and Evolutionary Studies
University of Groningen
PO Box 14
9750 AA  Haren
The Netherlands

Phone      + 31 50 363 2236
Fax          + 31 50 363 2154
Email        r.haanstra at biol.rug.nl






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