Replies

• Dorothy Clyde

  20 March 2008 | 10:26

  Hi Nisha,

  In my time in the lab, the most common reason for gels not running properly was due to the gel being made incorrectly; usually the gel had been absent-mindedly made up with water instead of running buffer (we’ve all done it at least once!)....

  If you’re sure your running buffer is correct, double check the percentage of the gel is correct (if it’s too high, nothing will resolve properly).

  If the gel is fine, it might be worth checking the composition of your marker (and/or loading dye).

  Good luck!

  Best,
  Dot

• Nisha padmanabhan

  20 March 2008 | 13:26

  Thanks dorothy. The gel i made was a SDS-acrylamide, and for both my resolving and stacking, i stuck to the standard compositions. I am wondering if my acrylamide has gone bad. Would you know the normal shelf life of acrylamide powder? I did make fresh acrylamide and deionised the solution, that still didnt help.

• Joao Goncalves

  21 March 2008 | 09:39

  Hello,

  In my lab we keep the acrylamide powder at 4ºC…it solved some problems in the past. Can’t you borrow some acrylamide from some other group so that you can check if that is the problem?

  Good luck.

  João

• Nisha padmanabhan

  24 March 2008 | 10:33

  I did that as well. I think the pH of the solutions arent what they are supposed to be because i see an entire phase moving in the resolving gel and the marker isnt getting resolved.