96-well PCR troubles

John Alsobrook ALSOBROOK at biomed.med.yale.edu
Wed Nov 25 16:34:05 EST 1992


In <9211252027.AA14226 at server.uga.edu> mcgraw%gandal.dnet at SERVER.UGA.EDU writes:

> 
> 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
... cool stuff deleted ...
> 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.
... cool stuff deleted ... 
> 				Al McGraw

Al, haven't used a 96-well machine personally, but have contact with a few
labs that do. Have you changed your vendor for plates? Are the plates a
different batch?  Since they aren't optimized for heat transfer in rapid
temp gradients, you might be experiencing lot-to-lot variation in the 
plates themselves.  Just a thought...

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