DEPC treated water in PCR

Klaus.Matthaei at ANU.EDU.AU Klaus.Matthaei at ANU.EDU.AU
Tue Nov 1 19:39:43 EST 1994


>In article <38jofj$e1a at cerberus-138.wustl.edu>, MOE at KIDS.WUSTL.EDU (AARON
>J. MOE) wrote:
>
>> 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.
>
>eric
>
>--
>*************************************
>*Of course it's not true...         *
>*But let's make the bastard deny it!*
>*                        LBJ (1948) *
>*************************************
>eric c. anderson
>anderson at pharmdec.wustl.edu
>dept. of molecular bio. and pharm.               (314)362-3963 (lab)
>washington univ. school of medicine              (314)862-2435 (home)
>660 s. euclid box 8103                           (314)362-7058 (FAX)
>st. louis, mo 63110

Be careful diluting primers, nucleotides etc in 10mM Tris 1mM EDTA.  The
EDTA mops up your magnesium and can kill your PCR.

I prefer 10mM tris, 0.1mM EDTA which works just as well.

Cheers, Klaus

*************************************************************************
Klaus Matthaei
Head, Gene Targeting                            |           |
The John Curtin School of Medical Research      |  _--_|\   |
The Australian National University              | /      \  |
Snail mail: Canberra, ACT 0200, Australia       | \_.--._/<<|
E-mail: Klaus.Matthaei at anu.edu.au               |       v   |
Landline: 61 6 249 3782
                        "Sometimes I'm the Louisville slugger"
                        "Sometimes I'm the Ball"
                                Dire Straits
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