Strand-specific 32-P probe by PCR????

neale at mbcf.stjude.org neale at mbcf.stjude.org
Fri Aug 18 15:51:53 EST 1995


Hello netters,


I would like some assistance in making 32-P labeled probes by thermal cycling.

I want to make a strand-specific probe to test on Northern blots. Yes, I can
clone and make riboprobes (and I am pursuing that also), but there ought to be
a quick way of using a single primer with a small amount of my insert DNA.

I wish to incorporate 32-P nucleotide into the nascent strand and not end label
the primer so that the specific activity will be comparable to random primed
probes.

I have checked a number of PCR protocol books, but they only offer advice on
the production of non-isotopic probes and not necesarrily strand-specific. If
anyone has a working protocol that they could share I would be very grateful.

Thanks,

Geoff Neale

Dept. of Virology and Molecular Biology        E-mail: neale at .stjude.org
St. Jude Children's Research Hospital           Phone: (901) 495-3400
Memphis, TN                                       Fax: (901) 523-2622





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