Southern Blot Background

George Rutherford gruther at bilbo.bio.purdue.edu
Fri Oct 9 15:31:43 EST 1998


In article <361CDB71.46B4BF8B at arches.uga.edu>, Brown Lab
<aedes at arches.uga.edu> wrote:

> Hello all,
> 
> I have been having background problems with the top half of my southern
> blot.  The bottom half of the gel is devoid of any background, however
> the top is completely black.  I  ran a second the gel without EtBr,
> thinking that may be the problem but I obtained the same background
> pattern.  I am using a positively charged nylon membrane, an 800bp  P32
> probe, and I hybridize and wash at 55C.  Has anyone else experienced
> this background problem?  Any help woud be greatly appreciated.  In
> addition, has anyone prepped genomic DNA from Aedes aegypti and
> succesfully used it for a Southern.  If so what protocol(s) did you use.


I trust you have excluded the possibility of a light leak?



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