Vent PCR Woes
brunstei at UNIXG.UBC.CA
Fri Jun 10 13:42:57 EST 1994
Are you overlaying your reactions with mineral/paraffin oil? If
so, try a different batch! I spent two months trying to get a simple
Vent PCR to go and in the end it all came down to the mineral oil I was
using. One clue was that with the bad mineral oil, while i was unable to
get any product bands under any of the conditions I tried ( [Mg++],
temp., +/- DMSO) I *did* see the primers dissapear from the reactions at
about what I thought should be the correct annealing temperature. If you
can see this, then you know that your polymerase and dNTP's are OK, which
can help to narrow down the field of possible causes.
Good luck in finding the answer!
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