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Drying agarose gels

James Kami jakami at ucdavis.edu
Mon Mar 22 14:14:38 EST 1999


A method I used many years ago came from a Hoffer techniques pamphlet.
Essentially, the gel has first denatured with NaOH as if preparing for a
southern blot, then dried onto standard filter paper using a vacuum gel
drier for one hour with no heat followed by one hour with low heat, less
than 55 degrees so as not to melt the gel. After drying the gel has
rehydrated in DI H2O and floated free of the filter paper. The result
was a thin, leathery material that was remarkably tough. It could be
used for hybridization just as a Southern blot on filter paper, although
stripping had to be done chemically rather than by heat. The great
advantage was that I could use small probes (20-40 bp) and had NO
background. The gel could be reused many times and still gave extremely
clean RFLPs. I used probes up to 500 bp with no problem.
	Depending upon the size of your protein, I wonder if you might not be
able to incubate a dried DNA gel with a protein solution and detect
protein binding useing an antibody or other method. My understanding is
that, unless the DNA-protein binding is of very high affinity, gel shift
sometimes doesn't work. Anyway, it was just a thought.

Good Luck

Jim Kami PhD.
Blue Rose Biotech
Davis, California 95616
Dr. Fernando Rodriguez-Pascual wrote:
> 
> I will be very grateful if someone can give me some tips to dry agarose
> gels. The purpose is to perform a kind of electrophoretic mobility shift
> assay to look the possible interaction between a 560 bp radioactive
> transcript and a purified protein. For this transcript size I should use
> agarose instead PAGE, but I was told that the drying of agarose gels is
> not easy.
> 
> Thank you in advance
> 
> --
> #############################################
> Dr. Fernando Rodríguez Pascual
> Department of Pharmacology
> Johannes Gutenberg University
> Obere Zahlbacher Str. 67
> D-55101 Mainz, Germany
> Tf. +49 (6131) 17 68 36 or 17 32 45
> Fax. +49 (6131) 17 66 11
> email: frodrigu at mail.uni-mainz.de
> #############################################



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