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!
> 
>
> Hi
> 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.
> Regards
> Simon Plyte
> splyte at ocicl.oci.utoronto.ca


>                    



More information about the Methods mailing list