PCR probes by incorporation of 32P ?
horton at biosci.cbs.umn.edu
Mon Mar 4 14:45:58 EST 1996
Geoffrey Neale (geoffrey.neale at STJUDE.ORG) wrote:
: Hello everyone.
: I am looking for a method for making 32-P labeled probes during PCR. The
: information that I specifically need is the concentration of cold
: nucleotide in the reaction so that you generate sufficient full length
: probes, but still retain high specificity of the isotope label.
: Other considerations are the amount of starting template, the
: concentration of primers, and the number of cycles.
: Thanks for your input.
: Geoff Neale
: PS Could you please email me your advice since our news reader is not yet
: operational on our system. E-mail is my only access at this time. Thanks.
(Yeah, I'll e-mail it, too. Posting here for the general audience...)
Schowalter DB. Sommer SS. The generation of radiolabeled DNA and
RNA probes with polymerase chain reaction. Analytical Biochemistry.
177(1):90-4, 1989 Feb 15.
You can achieve higher specific activities with lower total [dNTP],
but you need to increase the extension time.
Robert M. Horton
http://188.8.131.52 Have a :) day
"Scotty, try flushing the radioactive waste into the ventilation system!"
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