Help wanted staining DNA gels

wmelchior at fdant.nctr.fda.gov wmelchior at fdant.nctr.fda.gov
Tue Apr 26 13:05:54 EST 1994


In article <2p5qb5$qd7 at s.ms.uky.edu> jdf at seqanal.mi.uky.edu (Jackie Fetherston) writes:
>Subject: Help wanted staining DNA gels
>Date: 21 Apr 1994 12:08:05 GMT

>Does anyone know of a nontoxic alternative to ethidium bromide to
>stain DNA in agarose gels?  We had a high school teacher doing research
>in the lab last summer who would like to show his students some molecular
>biology but would like to avoid carcinogens.

A couple of years ago, I asked the same question for the same reason.

0.02%  (20 mg/100 ml H20) methylene blue works pretty well.  Place small (ca 2 
x 3 inches) gel in 100 ml stain and rock gently.  Let it sit 1-1 1/2 hr.  
Transfer gel to 1 liter H20, rock gently, and let sit overnight to destain.

I developed a procedure for plasmid isolation that I think would be suitable 
for high school -- uses a clinical centrifuge with regular 6 inch test tubes 
-- if the teacher can grow an overnight culture of plasmid-infected bacteria 
and has access to a  higher capacity, but still low speed centrifuge.  I only 
did it once, and it's never been tried in a classroom, but I'd be happy to 
make it available to those who would like to try it or use it as a starting 
point.  I know there are also similar experiments available from other sources.

Bill Melchior
wmelchior at fdant.nctr.fda.gov                                   (501) 543-7206 
_____________________________________________________________ 
Opinions stated are not those of NCTR or its sponsoring agencies.

National Center for Toxicological Research
FDA
Jefferson, AR  720729




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