hot glass plates from acrylamide gels

Klaus Salger salger at wap1.zi.biologie.uni-muenchen.de
Wed Mar 19 18:56:48 EST 1997


Timothy R Jackson (tjackson at uoguelph.ca) wrote:
: has anybody experienced the following problem and managed to solve it...

: we are using 6% denaturing acrylamide gels to separate pcr amplified 
: microsatellites.  one of our primers are end-labelled with gamma 
: 33P-ATP.  after running our gels, we fix them in a 10% methanol, 10% 
: acetic acid solution and then dry them.  we have recently found that when 
: we remove our gel from fixer and lift it off of the glass plate, the 
: glass plate that the gel was fixing on is radioactive in the same region 
: as the amplified microsatellite locus.  in some cases the glass plate is 
: hotter than the gel!!  this appears to be affecting the consistancy of 
: our results as some glass plates get hotter than others and could be 
: reducing the signal of our gel.  any solutions????


Tim,

I experienced the same problem once when doing gelshifts. Cleaning the
plates VERY thoroughly with ethanol and water followed by siliconization
fixed the problem for me.

Cheers
  Klaus

--
Klaus Salger		phone : +49 (0)89 5902 	-502
Zoologisches Institut	FAX   :			-450
AG Wetterauer		e-mail: salger at zi.biologie.uni-muenchen.de
Luisenstr. 14
80333 Muenchen
Germany

BioLinks: http://www.zi.biologie.uni-muenchen.de/~salger/salger.html



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