TBE precipitation

Joan Marie Shields jshields at taurus.oac.uci.edu
Thu Aug 12 14:32:27 EST 1999


 
Colin Rasmussen <colin at pombe.usask.ca> wrote:
>> >I think the borate has a tendency to ppt. with time.  We found in the
>> >past that autoclaving it helps for some reason.  Alternatively, make 5 x
>> >TBE.  That always solves the problem.  We typically run our gels in 0.5
>> >x TBE so you won't wind up making any more than you are now.

Joan Shields <jshields at uci.edu> wrote: 
>> I was having trouble with it precipitating as well - autoclaving seems to
>> have solved that problem.
 
>> I will say regarding the 0.5x - I typically run fairly high percentage
>> agarose gels (2-4%) and I've found that if I use 0.5x TBE that my bands
>> aren't nearly as sharp.  I've gone back to using 1x TBE and have had much
>> better results.
 
>What voltage do you run at.  We've found that at 100v on a minigel,
>sharpness is fine, although I must admit we don't run that high
>percentage.  When I need to separate smaller things I usually switch 
>to acrylamide.

Oh, usually between 60 and 90V, depends on the size of the gel.  One of 
the other fellows here in the lab has also had better results with 1x 
as opposed to 0.5x buffer.  I think it may depend on a variety of factors.
If 0.5x works for what you are running then stay with it - but if you are
seeing your bands become less sharp than they should be I would suggest
trying 1x TBE.



joan


-- 
Joan Shields       jshields at uci.edu       http://www.ags.uci.edu/~jshields
University of California - Irvine         School of Social Ecology
Department of Environmental Analysis and Design
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