PCR contamination

Shaun Tyler styler at HPB.HWC.CA
Thu Mar 21 20:06:38 EST 1996

> PCR contamination
> Neil Burns (pdxrnb at pdn1.gene.nottingham.ac.uk)
> Tue, 19 Mar 1996 10:33:16 +0000
> ---------------------------------------------------------------------=
> To: methods at net.bio.net
> From: pdxrnb at pdn1.gene.nottingham.ac.uk (Neil Burns)
> Subject: PCR contamination
> Date: Tue, 19 Mar 1996 10:33:16 +0000
> Hi All,
> Has anybody ever heard of or tried UV irradiation of primers for use
> in PCR
> to remove contamination?
> Cheers in advance,
> Any replies to e-mail please,
> Linz

You =93betchya=94.  We routinely work with a set of primers which are=20
universal for the 16S gene from eubacteria and have had extensive=20
problems with contamination.  To cure this we now UV irradiate our=20
reaction mixture (primers, dNTP=92s and all) before adding the Taq and =
template.  We have also done this with the Taq already added and it sti=
seems to work fine.  Our standard procedure is to prepare the reaction=20
mixture and then put it through 5 =93Auto-crosslink=94 cycles (1200 kJ =
in our crosslinker (from Stratagene) with vortexing inbetween each cycl=
 I have heard that other people simply place their reaction mixture on =
transilluminator for a few minutes with equal success.  Give it a try. =
I=92m sure it will solve a lot of your headaches.


Shaun Tyler
Head, DNA Core Facility
Laboratory Centre for Disease Control
Health Canada

Ph#:  (613) 941-6441
FAX#: (613) 957-1358

E-mail:  styler at hpb.hwc.ca

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