NEW Sequencing Gel Problem

Greg Hawkins hawkins at MAP.MARC.USDA.GOV
Tue Mar 2 12:46:06 EST 1993

        Our lab needs some helpful explanations for our new sequencing gel
problem.  We are very experienced with sequencing using the Sequenase kit
and AT-Biochem's Long Ranger gel (5%).  Our problem: When we run our gels
for 1.5 hr (short run) our sequencing is perfect.  On the SAME gel we run
the same samples for 3 hr (long run) and get smears (no banding pattern at
all).  We control the temperature of the gels carefully.  We have run
hundreds of samples like this in the far and recent past with no problem.

        Our conditions:
               - M13 
               - Sequenase kit
               - NEN S-35 dATP
               - 40 cm gel; 5% AT-Biochem Long Ranger
               - 1 X TBE
               - 55 Watts

        Is ther something wrong with this batch of Long Ranger??  Could it
be a running buffer problem??  Please respond soon!


                hawkins at (Greg Hawkins)

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