Cleaning contaminated primer stocks (SUMMARY)
Simon E Plyte
splyte at ocicl.oci.utoronto.ca
Fri Nov 26 09:28:36 EST 1993
In article <1993Nov26.101544.2782 at infodev.cam.ac.uk>,
mjl17 at mbuc.bio.cam.ac.uk (Michael Lush) wrote:
> Last Friday I posted a plea for help Re: Contaminated
> Primer stocks. I recieved lots of email help which I have edited
> down a bit and am now posting with a big thankyou to everyone who
> sent advice....
> Also just as a little postscript the primer stocks
> wern't contaminated (praise be!!), it was probably one of
> the buffers... Anyway thanks again for the advice, I just hope
> I never need it!
> I have just joined the Net and read about the problems of primer
> contamination. I had this problem a couple of years ago and solved it
> in a couple of ways: a) UV light b) DNAse treatment.
> A) UV treatment... this was done by irradiating the samples in a
> UV stratalinker at 254 nm for 10 mins. (ref. Sarker and Sommer, 1990,
> Nature 343 : 27)
> B) DNAse treatment... Here I added 0.5 units of DNAse to my PCR reaction
> mix (i.e. everything added except the template and taq) and incubated
> at RT for 10 mins....then boiled for 10 mins to kill the DNAse prior
> to the addition of template and taq. I never tried adding the DNAse
> straight to the primer stock, but doing it this way eliminates the
> contamination from all of your stocks. (Ref. Furrer et al., 1990,
> Nature 346: 324).
> I always do the DNAse treatment these days....just in case.
> Anyway, glad to hear you primers weren't contaminated.
> Simon Plyte
> splyte at ocicl.oci.utoronto.ca
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