Big Dye blobs with AGTC plates

Margaret Scott mscott at rascal.med.harvard.edu
Fri Dec 5 13:05:54 EST 1997


We are using Big Dyes and had great success with them while using
Centricept Columns to process the sequencing reactions. Since we changed
to the AGTC 96 well format, the dyes are not being removed and show up
as bright yellow blobs at the beginning of each lane (obscuring the
sequence).  The 96 well format saves so much time, I hate to go back to
individual columns.  Has anyone found a gel filtration product in a 96
well format that removes the Big Dyes?  I have talked extensively with
AGTC and they have suggested some changes in the spin time and force
during the centrifuge step, but that has not aleviated the problem. 
Thank you in advance for any insite you can provide.

Margaret Scott
Biolpolymer Facility
Harvard Medical School




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