taurine gels

Wolfgang Schechinger wgschech at med.uni-tuebingen.de
Tue Sep 1 04:11:48 EST 1998


Seems to be a question a of temeprature. At least I observed this 
when pre-running sequencing gels too long. The cause probably is 
expansion of the gel.
You might try running the gel at lower temperatures (=less voltage) 
or in the cold room. If it's possible to decrease the mechanical 
force that clamps to two glass plates together in the loading region 
of the gel, this could help, too. 

Just some ideas. 

Wolfgang

> We are trying to use taurine gels for manual sequencing with new
> thermosequenase and ATP-p33. We got some trouble in the loading
> surface after 2 hours run at 50oC, that seemed to be uneven and
> irregular, making the second loading impossible.    Conditions used
> for the gel: 6% acrylamide, 8M urea, 1 X GTG buffer, 1 ml APS 10%,
> 25 microlitres of TEMED (for a 100 ml gel). All itens are from
> Pharmacia except for GTG buffer:USB mixed powder . These conditions
> were suggested at the thermosequenase protocol by Amersham. We also
> tried TEMED and  APS 25%, 100 microlitres each, and the problem
> persisted.
> 
> 
> Does anybody would have a suggestion to overcome those problems?
> Please help me! I would appreciate very much any sympathetic
> message. 
> 
> 
> Mario Mayer
> Instituto de Ciencias Biomedicas
> Universidade de Sao Paulo
> Brasil
> e-mail : tefcosta at usp.br
> 
> MZÉ
> 
> 
-----
usual disclaimers apply * This message is RNAse free - please don't touch!
-----                              
Wolfgang Schechinger         
University of Tuebingen, Germany
email: wgschech at med.uni-tuebingen.de * wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
-------



More information about the Methods mailing list