DNA staining

Donald A. Lehn donnel at helix.nih.gov
Tue Jul 30 11:21:26 EST 1991

In article <910729200935.20400736 at NTDOC.NCTNET.GOV> WMELCHIOR at NTDOC.NCTNET.GOV writes:
->I am helping develop an experiment for high school students
->involving agarose gel electrophoresis of DNA.  My question is,
->what stain should we use.  I have read that methylene blue
->can be used -- this would avoid the use of the mutagenic
->ethidium bromide and of ultraviolet light, which would be
->desirable for working with a group of students.  On the 
->other hand, I'm worried that the sensivity would not be 
->high enough.  I'd like to keep to less than a microgram
->of DNA per band.
->Does anyone have experience with methylene blue staining
->of agarose gels?  I haven't been able to find anything
->in my "standard" reference works about how to use it or
->how sensitive it is.  Or are there other non-hazardous
->stains I should investigate?  (I need advice on staining 
->solutions, such as dye concentration and buffer components,
->as well as destaining solutions and procedures.)
->I'm anxious to help my teacher friend develop an
->exciting but safe experiment.  Any help will be much
->Bill Melchior      
->National Center for Toxicological Research
->Jefferson, AR  72079
->(501) 543-7206
->wmelchior at ntdoc.nctnet.gov

You might consider using biotinylated DNA.  You could then combine with
this experiment the use of enzymes (avidin coupled with peroxidase) to
tag molecules.

Don Lehn

Donald A. Lehn, Ph.D.                      Phone: (301) 496-2885
Bldg.37 Rm 3D20                            FAX:   (301) 496-8419
National Cancer Institute / NIH            Email: donnel at helix.nih.gov
Bethesda MD 20892

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