Non Radioactive Southern Blotting - Teaching

Mike Prigge prigge at darkwingREMOVE.uoregon.edu
Fri Oct 23 04:10:34 EST 1998


In article <362F2407.8954CA7B at rpms.ac.uk>, mjones at rpms.ac.uk wrote:

> Hi
> 
> I run a MSc course, and the students carry out various mol biol
> techniques, in particular a Southern blot, Northern blot and lambda
> phage library lift screening.
> 
> Up until now I have used P32 to make radioactive probes to screen the
> filters for the above techniques.
> 
> I was wondering if anybody could point me in the right direction (if
> considered suitable) for a reliable non-radioactive method of detection
> (Amersham ECL, biotin, digoxigenin, etc) to be used with the above three
> techniques.
> 
> Many thanks
> 
> Could people reply direct to my email address (as well as the news
> group).
> 
> mjones at rpms.ac.uk

I TA'd a molecular biology lab 4 years ago when I was a U of Minnesota. 
The class used BMB's Genius Kit (digoxigenin label and CSPD detection) to
do a Drosophila genomic Southern blot for RFLP mapping.  It worked quite
well.  However, there MIGHT be a problem with plaque lifts on
nitrocellulose (instead of nylon) membranes.  I now successfully use
nitrocellulose membranes but I switched to the anti-DIG-Peroxidase
antibody and luminol detection instead of anti-DIG-AP and CDP-Star.  I
cannot comment on whether it would have worked with anti-DIG-AP nor can I
comment on Northern blots since I have never tried them with the Genius
system.

Good Luck,
Mike

-- 
Michael Prigge                          Phone: (541) 346-4256
Institute of Molecular Biology            Fax: (541) 346-5891
University of Oregon /\\ /\\ /\\ /\\ /\\ /\\ /\\ /\\ /\\ /\\
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