DEPC treated water in PCR

Enrique Jose Labadan Frio - SCBC - 3421101 g3421101 at MUCC.MAHIDOL.AC.TH
Wed Nov 2 01:30:38 EST 1994


> > Does anyone know if DEPC-treated water will cause problems in PCR?  I just
> > found out that some of my oligo primers were diluted in DEPC water and they
> > don't seem to work anymore.  Thanks.
> 
> aaron,
> 
> the problem that you may be having is not with the DEPC, but with the water
> itself.  primers which are diluted in water rather than Tris or TE (my
> personal favorite) break down much more quickly than if they are in a
> buffered solution.  FWIW, i always elute, dilute or whatever-lute DNA
> (oligos, cDNAs, genomic, etc.) in TE (10mM Tris, 1mM EDTA pH8) and end up
> having much better luck with them.

	Just curious - but EDTA can and will chelate some of the Mg2+ in
	the PCR mix and skew the yield or specificity somewhat. Depends on
	how much ul of primers [depends on concentration] are added to the
	mix. 

	There was a previous post a long time ago stating that trace
	amounts of DEPC inhibits Taq pol, and this may be the source of
	the problem. 

	Also, why would primers "break down" in clean water? If DNA could 
	survive chemical and environmental harshities and be used in
	forensic analysis, then....

	AIMS? [Am I missing something?]


Disclaimer: I speak only for myself.

============================
Querix
g3421101 at mucc.mahidol.ac.th
============================
Scientific snigglet [words that should be in the 
dictionary, but aren't]:

"squiggles" = those little bits of gel stuck to the comb after pulling out 
	      from a freshly-made gel



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