EtBr staining acrylamide gels

holin sue holin at pc2.hinet.net
Tue Apr 23 11:17:26 EST 1996


>
>Pete wrote:
>> 
>> > > Does anyone know if it is possible to incorporate EtBr dye into a
>> > > polyacrylamide gel while casting so that the DNA stains is stained as
>> > > it is run?
>> > >
>> I've stained the gel afterwards by incubating it in 1 ug/ml EtBr for some
>> time, haven't tried adding it to the gel, though ...
>> 
>
>Well, i have done it with agarose gels, not PAGEs.  i would imagine that
>it would probably work just as well. it works extremely well with
>agarose gels like i mentioned before(with a very few minor exceptions).
>i add 0.5 ul for every 10 ul of agarose gel.  so that would be 
>2.5 ul/50 ul gel. this works very wel.  th only problem would be during
>purification form the gel.  the et.br intercalates into the dna and if
>you want super, duper clean dna then you might consider somehing else.
>of course, the et.br intercalation factor is extremely negligible until
>unless you are doing dna curvature studies or protection/binding
>assays...cheers... 
>
>
>
>> Pete
>
>-- 
>Arioch (sai at topaz.microbio.uab.edu)
>Daemon prince of Chaos (a.k.a Cell and Molecular Biology Graduate
>student)
>The East side of the 9th plane of hell (a.k.a Dept. of Microbiology)
>Dimension of Chaos (a.k.a University of Alabama at Birmingham...)
>
holin(national defense of medical college)
usually I stain the acrylamide gel in the concentrated EtBr after the
gel was run off. The gel must destained in water for several hours. Then
you can see what you want to see.



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