Help wanted staining DNA gels

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Thu Apr 21 15:00:07 EST 1994


 In article <2p5qb5$qd7 at s.ms.uky.edu>
 jdf at seqanal.mi.uky.edu (Jackie Fetherston) writes:

| 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.

I suppose I should include this in the FAQ list. A while ago, people were
discussing this very issue. The best posts are found below.

================================================================================
> As part of a followup to these two methods of subcloning agarose
> gel purified DNA fragments I have found that using methylene blue
> to visualize the bands vs. EB UV light results in a 10-fold higher
> efficiency in insert-containing clone isolation.  Simply stain
> your agarose gel in 0.25% methylene blue in 0.1X TAE for 30 min
> and destain in 0.1X TAE until you can see the band you want.
> The only contraindication I know for this method is that it doesn't
> work with a product called Synthagel.
>                                    Eric
> -- 
>   Eric R. Hugo, Postdoctoral Research Associate |eric at bcserv.wustl.edu
>   Dept. of Biochemistry and Molecular Biophysics|                
>   Washington University School of Medicine      |               
>   Box 8231, St. Louis, MO 63110_________________| (314) 362-3342
================================================================================

================================================================================
> Recent edition of TIG (9(2):pg 40) reported comparison of basic dyes incl.
> methylene blue for nontoxic staining of agarose gels.  They found the best was
> to immerse the gel in 0.04% brilliant cresol blue in 20% ethanol for 15-60
> mins, wash dH20 10s, 70% ethanol 5s, twice more dH20 to detect 25ng of DNA.
> 
> Haven't tried it, but they reckon lower background than methylene blue
> 
> Cheers
> 
> Brent Gilpin
> Crop & Food Research
> Lincoln
> New Zealand
================================================================================

That should do it. Warning: One person reported that the use of brilliant cresol
blue resulted in a completely blue gel. Maybe you should try it first before
demonstrating it to the entire high school class, else egg looks good on the
teacher's face. Good Luck!

-Paul.

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* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
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