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UV sterilization of PCR reagents

UV sterilization of PCR reagents - Protocols and Methods Forum

UV sterilization of PCR reagents - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 10-20-2004, 06:01 PM
Charlie Butterworth
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Default UV sterilization of PCR reagents



Hello,

I am setting up a new molecular lab, and will not have access to Type I
water for reagents. I can filter sterilize most of the reagents, and /
or use purchased Type 1 water.

However, I am thinking that small quantities of water that would go
into reagents for DNA stocks (such as TAE for DNA stocks, and water for
dissolving new primers) could be aliquoted into 1.5 mL tubes then
exposed to UV light from a UV lamp (mineralight). Even PCR cocktails
(minus Taq, primers, template and mineral oil) could be UV sterilized
in PCR tubes.

OK, so are PCR tubes and 1.5 mL plastic tubes UV permeable?

If so, which wavelengths of UV are best?

I'd appreciate any advise.


Charles A. Butterworth (Ph.D.)
Research Specialist
Desert Botanical Garden
Phoenix, Arizona

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  #2  
Old 10-20-2004, 06:29 PM
Wolfgang Schechinger
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Default UV sterilization of PCR reagents

Charles,

you'll probably kill yll your reagents. UV induces lots of radicals and
specially will destroy your bases.... That's the way you're actually
killing the bacteria in your mixes. Better spend a few bucks on Molecular
Biology Grade Water (a sixpack of 100 ml bottles is about 100 USD, aliquote
and frezze down 1 ml aliquots to avoid contamination of your stock). Or
stick at least to RO "distilled" water from local pharmacy or gas station
(for batteries). If you don't have an autoclave, sterilize it in a pressure
cooker or use 0.1 um syringe filters.


At 19:01 20.10.2004 +0100, Charlie Butterworth wrote:
Wolfgang Schechinger, PhD (Dr. rer. nat.)
Chemistry and Life Sciences

BGFA XU44
Med Clinic I / Endocrine Research
University Hospital "Bergmannsheil"
Bürkle-de-la-Camp-Platz 1
44789 Bochum
Germany

Phone +49-234-302-4789
Fax +49-234-307-4790

[Only registered users see links. ]




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