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sequencing

Gabrielle E. Rieckhof ger4 at merhaba.cc.columbia.edu
Tue Dec 7 21:23:08 EST 1993


Dear bionetters,

This is not a yeast question, per se, but I am sequencing a yeast gene. Anyway,
when I sequence, I routinely load one set of sample, and load a second set 
after the xylene cyanol marker of the first set has run out of the gel. When I
am ready to load the second set, however, the top of the gel (the part that the
comb touches to form the wells) has become distorted. I do not know if this is
due to the heat or the electrical current, but it interferes with loading of 
the second set of samples.

I've found that running the gel on a lower wattage helps, though not 
completely. I am using a wedge spacer on a 6% gel, and running at 50 or less 
watts (about 1200V and 40mA). 

Thanks in advance for any and all help.

KC.

P.S. Another problem I have is that the gel also developes air bubbles several 
centimeters from the top after extended runs. Any remedies?



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