Hi, I tried washing the wells with distilled water and running buffer using syringe and extended tip prior to loading to wash off any unpolymerized acrylamide. It stacked to the side and bottom of the well. I used same size spacer and comb.
Any help would be greatly appreciated.
Nishan
Im presuming that you tried to cast a discontinuous gel and faced the problem which usually happens when there is air space between your comb and the acrylamide polymers. Most common reason is that you filled the space (for Stacking Gel) partially and then inserted comb with expecting it will cover the entire space. However, with even minimal gap, the issue crops up. Secondly if your spacers and comb are old and Teflon comb is bent, it leads to small air pockets and same issue. Always use spacers and comb from same lot.