Help with EMSA minigel conditions

Darren Tyson tysondr.nospam at slu.edu
Thu Oct 16 09:57:19 EST 1997


Howdy,

I'm trying to convert my electrophoretic mobility shift assay (EMSA)
from a large gel format (20 cm X 20 cm) to a minigel format (BioRad
Mini-Protean II).  For the large gel format the running conditions were
constant 35 mA at 4 deg C for 4 hours.  I believe constant 35 mA for a
minigel would be too much power for the protein-DNA interactions to be
maintained and would finish the run very quickly.  Can anyone suggest
the proper running conditions for my application?  BTW, I'm using a high
ionic strength buffer (Tris/glycine/EDTA).

Any help would be greatly appreciated.  Thanks in advance.

Darren




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