RNA blot question

Heinz-Juergen Schaefers mfm434 at rzmail.uni-erlangen.de
Mon Jul 29 15:16:15 EST 1996


Julie Roberts wrote:
> [skip]
>
> We then attempted to use more specific mouse probes
> constructed in our lab using PCR  and then began the trouble.
> 
> One blot in particular looks as though it has a reverse hybridization
> pattern in which the probe has hybridized to everything but the RNA
> bands. Has anyone seen anything like this before? Can anyone explain
> this hybridization pattern?

Hi!

I got this "reverse hybridisation" one or two times, using a very short
PCR fragment (230 bp) as radiolabled northern probe. The "reverse hybridisation"
band was at height of the 18s rRNA.
The reason for many unspecific bands or high background is probably a too short 
PCR fragment as probe.
The reason for this "reverse hybridisation" i don't know...

bye
	Heinz
-- 
Dipl. Biol. Heinz-Juergen Schaefers
University of Erlangen, Med. IV, Nephrology Research Lab
E-mail: university: mfm434 at rzmail.uni-erlangen.de
           private: h-j.schaefers at erlangen.netsurf.de



More information about the Methods mailing list