Help spotty northern problem help

Juergen Brosius brosius at uni-muenster.de
Wed Aug 6 10:35:29 EST 1997


In article <33E76F50.3D1E at leleand.stanford.edu>, punk at leleand.stanford.edu
wrote:

> Hi, I am doing a lot of northerns on cellular RNA and having a problem
> with spots on my autorads. These spots are small and round. The
> hybridization signal is good, no crosshyb to rRNA and the general
> background is low except for the spots. I am using  zeta-probe membrane
> from biorad and have changed lots which didn't help
> 
> - I am using the .5M NaPO4  7%SDS  hyb buffer. It used to work well no
> spots and this is what is recommended for the zetaprobe.  I have switch
> the solutions and filtered them but still the spots are present.
> 
> - I am using random primed probes and a hybridization oven. The probe is
> cleaned up using qiagen spin column 
> 
> If any body has any suggestions on what might be causing these spots
> please post to this group I check it regulary Thanks alot gregg

Do nont use powdered gloves  -- good luck !

Juergen

-- 
                                          usual disclaimers
Juergen Brosius
Inst. f. eXp. Pathol.
University of Muenster
Von-Esmarch-Str. 56
D-48149 Muenster, Germany

Fax: +49  251  835 8512
Tel: +49  251  835 8511



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