96-well PCR troubles

mcgraw%gandal.dnet at SERVER.UGA.EDU mcgraw%gandal.dnet at SERVER.UGA.EDU
Wed Nov 25 15:27:39 EST 1992


Hi netters,

Here's the problem.  We have a PCR that we've used many times on many
different DNAs.  We feel that we have the conditions pretty well
optimized.  It has worked consistently both in a 60-tube format and 
in a 96-well microtiter tray format.  We have both machines from M-J.
The PCR is a "microsatellite", or dinucleotide repeat polymorphism, with
two twentymer primers.  Recently, the reaction stopped working for us,
but only in the 96-well machine. If you mix up a reaction, split it, and
run half in a tube and half in a microtiter well, the tube rxn works, the
other doesn't.  This makes us suspect the machine, but as nearly as we
can tell by watching it, the machine appears to be operating properly.
Has anyone out there had a similar problem with microtiter PCR's in an
M-J machine?  What else might be the problem?  We use Falcon plates.
Any comments appreciated.  Thanks.

				Al McGraw



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