User:Paul Rothenberg/Notebook/CHEM 571 Fall 2014/2015/04/15
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SDS Page Gel Electrophoresis
I. 4 - 12 well pre-cast Mini Protean TGX gel was obtained and prepared.
II. The Bio-Rad Mini Protean system electrophoresis cell was assembled.
III. Each of the 4 gels were loaded with each of the 20 μL reaction samples prepared yesterday in the following manner:
IV. The gel was run for approximately 10 min at 200 V, 0.05 A, and 10 W.
NOTE: The top of the chamber was not functioning properly in that a closed circuit was not established and the top had to be manually held/pushed down for the entirety of the run.
V. After 10 min, the gel was placed in a Fixative Solution (40% methanol, 10% acetic acid, 50% water) for 30 minutes
VI. The gel was then placed in a Stain Solution (0.025% (w/v) Coomassie Blue, 10% acetic acid, 90% water) for approximately 24 hours
VII.Finally, the gel was placed in Destain Solution (10% acetic acid, 90% water) three times for 15 minutes each and the following results were obtained: