2D gel running problem

[cosmis85]

Hi I've been having a certain problem when running my 11cm 2D gels in a Criterion gel tank by Bio-Rad. I run them under constant voltage conditions of 200V but I seem to get really low mA values. As a result my gels will either not run at all giving me a total smear when stained or stop running midway through. Additionally it takes 2-3 hours for them yo run as opposed to 50 min previously. Lately I have been having the same problem with my 1D gels as well. From that I thought that the problem was with my electrical source ( PowerPac 300) but I switched it with another one that we had in the lab but still have the same problem. Any ideas on what maybe at fault or what to do?

[apsbabu]

Check your Buffer .. I think there is too little salt in it !

[leaf]

I think the problem lies in you buffer,prepare a new buffer and do it again.

[Newscience]

The first problem is usually the powersupply however you should try a ready made buffer to eliminate buffer problems. If the problems still exist try running your proteins on a different system. I highly reccomend the Gebagel System from chembio Ltd.

[Only registered users see links. ]

[prot]

Do you prepare your gel correctly? Check if you use old acrylamide. Make sure you never recycled your buffer, use fresh for every run.