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.
Did you mix your running gel solution properly before pouring?
what are your samples and how do you prepare them. salts, lipids, etc, can cause your sample to run different from ideal.