ETBr in Loading Buffer

David L Haviland David.L.Haviland at uth.tmc.edu
Tue Feb 5 09:23:08 EST 2002


> I was wondering if anyone has ever tried or knows why Ethidium Bromide
> is not added to Loading buffer? I've never run across an article where
> Ethidium Bromide is present in the Loading Buffer, it's allways
> present either in the gel or as a post electorphoresis step.

Hi... 

ALL THE TIME... I put EtBr in the gel (2-4ul/100mls) and I have it in 
my loading buffer at 1ug/ml.  I never use it in the running buffer as 
we can't pour the buffer down the drain otherwise.   It has to be 
filtered through a EtBr reclaimation filter.  I've never had a problem 
in doing this over the last 10 years, neither in the running of nor in 
the interpretation of the data.  In fact, it was even a suggestion out 
of BioTechniques, a millenia ago... I can't even remember the issue.   
I even do it for northerns too but drop the amount in the gel to 1-
2ul/100mls of gel, lest the formaldehyde turn the gel into a "pink 
flare" on the transilluminator.

Clear skies...
David 

============ 
David L. Haviland, Ph.D., Asst. Prof. Immunology 
University of Texas - Houston, H.S.C. 
Institute of Molecular Medicine, R907 
2121 W. Holcombe Blvd.,  Houston, TX  77030 
713.500.2413-Voice//713.500.2424-FAX 
----------------- 
If everything seems to be going so well, you have obviously 
overlooked something. 
============

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