taurine gels

Carlos A.Moreira-Filho cmoreira at BIOMED.ICB2.USP.BR
Mon Aug 31 15:45:45 EST 1998


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
  
MZ



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