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10% acrylamide/bisacrylamide, or 15% ? - SDS PAGE gel making (Jun/29/2008 )


My SDS PAGE has been not doing so well. At the end of my electrophoresis, the lane width of the my SDS-PAGE Broad Standard (Ladder) narrows as the dye front passes the stacking gel into the running gel. I'm thinking I need to add less of the 12 % Acrylamide:Bisacrylamide into something like 10%. Does this sound okay? Will this help me get more rectangular bands and less "cone" shaped narrowing?



ps: another way of describing my problem is that i'm not getting a nice uniform lanes shape in my running gel after a 1hr 15 min SDS PAGE run.


Just wondering...

Did you mix your running gel solution properly before pouring?

-Minnie Mouse-

what are your samples and how do you prepare them. salts, lipids, etc, can cause your sample to run different from ideal.