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salt-gradient sequencing gels

Michael Myers myersm at rockvax.rockefeller.edu
Mon Nov 20 23:12:35 EST 1995

In article <48qg6p$1fnu at bigblue.oit.unc.edu>, jtsuruta at med.unc.edu (James
Tsuruta) wrote:

>How are salt-gradient sequencing gels run?  I saw an old post that said:
>0.5X TBE in upper resevoir
>1X TBE in lower resevoir + "3M NaOAC"
>I'm assuming that the concentration of acetate in the lower resevoir is
>less than 3Molar. (maybe 0.3M?) Can anyone tell me how they do this?

I do this all the time and it works well. However, I've always put 1X TBE
in the top and 0.5X in the bottom. Perhaps someone steered me wrong on
that, but it still works. 

You add the 3M NaOAc to the bottom reservoir to 1M final (ie. 500 ml of
0.5X TBE + 250 ml 3M NaOAc). If you pre-run your gels, don't add the NaOAc
until *afterwards*. You will see that the BromoPhenol Blue dye migrates
very slowly at the bottom of the gel.

Michael Myers
Laboratory of Genetics
The Rockefeller University
212-327-8233  (-7420 fax)
myersm at rockvax.rockefeller.edu

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