PCR contamination

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


> PCR contamination
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> Neil Burns (pdxrnb at pdn1.gene.nottingham.ac.uk)
> Tue, 19 Mar 1996 10:33:16 +0000
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> ---------------------------------------------------------------------=
-
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> To: methods at net.bio.net
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> From: pdxrnb at pdn1.gene.nottingham.ac.uk (Neil Burns)
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> Subject: PCR contamination
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> Date: Tue, 19 Mar 1996 10:33:16 +0000
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> Hi All,
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> Has anybody ever heard of or tried UV irradiation of primers for use
> in PCR
> to remove contamination?
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> Cheers in advance,
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> Any replies to e-mail please,
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> 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 =
DNA=20
template.  We have also done this with the Taq already added and it sti=
ll=20
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 =
each)=20
in our crosslinker (from Stratagene) with vortexing inbetween each cycl=
e.=20
 I have heard that other people simply place their reaction mixture on =
a=20
transilluminator for a few minutes with equal success.  Give it a try. =
=20
I=92m sure it will solve a lot of your headaches.

Shaun.

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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