Water used in PCR

David L. Haviland, Ph.D. HAVILAND at KIDS.WUSTL.EDU
Sat Jun 4 00:05:57 EST 1994


In <0097F67A.06A308E0.17 at umbsky.cc.umb.edu> 4700gbera at UMBSKY.CC.UMB.EDU writes:

> Hello Netters-
> 
> I'm having real trouble consistantly amplifying an 1880 bp fragment
> from bacteria. My first couple of trys worked beautifully. Then the 
> product pretty much disappeared. (except for one run a few days ago,
> this is really driving me nuts)  I've gone through the usual suspects -
> Taq, primer, 10x, Mgcl, etc etc. Right now I'm running a temperature 
> recorder on our thermocycler. I'm also wondering about our water supply. 
> 
> My question - what kind of water do you folks out there use for PCR??
> Has anyone run into any problems with it?

Gina:

In a word, yes.  We have found pure water to be critical not only in PCR 
but in libraries as well.  With either a Mili-q or Barnsedt (no 
endorsements here, these are the only two we've used...) as long as we get 
16-18 meg-ohm water our reactions seem to work well.  What we do is bake a 
flask (not autoclave yet), cool it, fill with pure water one of these 
machines, autoclave, cool, aliquot and freeze.  Treat it like gold and use 
only for *the* most important of reactions, like long (>1500 bp) PCR 
reactions or making libraires.

Hope this helps,
David




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