Vent PCR Woes

John Brunstein 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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