BRL RNA ladder hybridizes to my probe
ryan at mbcf.stjude.org
ryan at mbcf.stjude.org
Fri Aug 18 14:24:35 EST 1995
In article <DDH3B2.FEr at ns1.nodak.edu>, stallard at badlands.NoDak.edu (Lana C Stallard) writes:
> significant hybridization with a mouse DNA probe (a cDNA probe of
> angiotensin converting enzyme) to the ladder when i transfer the ladder
> onto a Northern blot. Why would there be so much sequence similarity
> Lana Stallard
Hi Lana,
When I've done RNA probe/DNA blot hybridizations, I've sometimes seen
uncceptable levels of nonspecific background hybridization which was fixed by
a wash in a single-strand-specific nuclease (I used RNase A). Probe must be
specifically & completely hybridized to be protected from digestion; non-
specifically hybridized counts come out in the wash.
Just a thought.
Good luck,
Kevin.
--
--
==========================================
Kevin W. Ryan
Department of Virology & Molecular Biology
St. Jude Children's Research Hospital
Memphis, Tennessee 38101-0318, U.S.A.
phone: (901) 495-3411
fax: (901) 523-2622
Internet: ryan at mbcf.stjude.org
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