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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