SUBJECT: Non-toxic DNA staining
DATE: 9/96
Biolabbers:
I have 2 questions:
Is there a procedure for qualitative analysis of nucleic acids that is
not dangerous nor too expensive ? Diphenylamine went on the district's
"forbidden" list and we are looking for another procedure.
Thanks in advance for your help
=============================================
Steve Uyeda
Mountain View High School
3901 W. Linda Vista Blvd.
Tucson, Az 85742
(520) 682-9190 - work (520) 318-3637 - fax
Steve,
Carolina Biological sells something called Carolina BLU for staining
agarose gels. Their catalog gives no indication of the sensitivity or how
many gels could be stained with the amount supplied for $4.95 (Cat#
K3-21-7300). Promega and Bio-Rad both sell silver staining kits but
they're fairly expensive and use chemicals that you probably aren't
supposed to use (Sodium thiosulfate, formaldehyde, silver nitrate).
Bio-Rad also sells amido black, which is supposed to work for DNA but
I've only ever used it for protein. Methylene blue works but is not very
sensitive. I also remember receiving something a year or two ago about
some non-toxic green stain, but I don't remember who sells it. Good Luck
Jeff
Jeffrey D. Newman newman@lycoming.edu
Department of Biology http://www.lycoming.edu/~newman/
Lycoming College Phone: 717-321-4386
Williamsport PA 17701 Fax: 717-321-4073
Hello BioLabbers-
I just read an article about visualizing DNA in agarose gels using
certain non-toxic dyes instead of ethidium bromide (I apologize if this
has already been discussed but I haven't been following all the threads
lately). Of several dyes that can be used, the article discussed the
use of Nile Blue and how it gave sharp and persistent bands and the fact
that it is not toxic. Nile Blue can be purchased from Sigma.
For those of you interested in the Nile blue article - it just came out
recently (two weeks ago) and it seems to be a sound article however, I
have not tried using the dye myself. The article came
from Analytical Biochemistry 240, 17-23 (1996). It is titled
"Visualization of DNA in Agarose Gels as Migrating Colored Bands:
Applications for Preparative Gels and Educational Demonstrations" by
Steve Adkins and Margit Burmeister.
Barbara Stegenga
***********************************
Barbara D. Stegenga
208A Coker Hall
Biology Department
UNC-Chapel Hill, NC 27599
Davidsoh.Coker@mhs.unc.edu
(919)966-3180
FAX:(919)962-1625
We have been using methylene blue, but
it does not have the sensitivity needed for the new labs. I really don't
want to be responsible for 150 students using ethidium bromide. I have a
technique for using brilliant cresyl blue (available through Sigma) I have
not had the opprotunity to try it yet. But as soon as I do I will let you
know how it works, if anyone is interested.
The reference for using brilliant cresyl blue is:
Trends in Genetics, February 1993 vol 9, no 2
The Article is " A Novel Stain for DNA in Agarose Gels"
Kirsten Mahovlich
Dept. of Biology
University of Toledo
We use Carolina Blu stain to visulize DNA and have been pleased
with the results. For $4.95 you get enough stain to run approx.
10 mini gels. It is a good deal and you don't have to deal
with ethidium bromide!
Nancy McInerney
Red Deer College
Alberta, Canada