Smiles in agarose gels

marty marty
Tue Feb 20 09:41:51 EST 1996


In article <landryp.824780576 at alize.ERE.UMontreal.CA>, 
landryp at ERE.UMontreal.CA says...
>
>Hello everyone.
>
>        In order to separate DNA fragments obtained from RAPD, I make 
the 
>amplification samples run through an horizontal agarose gel. 
>Unfortunately, I always get this "smile" that bends all the bands and 
>makes the gel analysis much more complicated. The gels are 1,5%, and 
I've 
>tried different voltages from 20 to 80 volts, and although I noticed 
that 
>the "smile" is reduced with decreasing voltages, it isn't clear enough 
to 
>allow a perfect analysis. I'd be glad to hear any suggestion, advice 
or 
>magic trick to improve the situation.
>
>Thanks in advance.
>
>Pierre-Alexandre Landry
>Dept. des Sciences Biologiques
>Universite de Montreal
>C.P. 6128, succursale centre ville
>Montreal, Quebec
>H3C 3J7
>we had the same problem running RAPD reactions out on gels.  Adding a 
small amount of EtBr to our ficoll loading dye (1µl EtBr stock 
(10mg/ml) per 1ml ficoll).  I'm assumming that each band within each 
lane is 'smiling' and its not just one big smile across the whole gel. 
Hope this helps.




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