PAGE electrophoresis - (Oct/11/2001 )

We have developed a suddem problem in the lab. The PAGE gel just refuses to set. Everything was working well till the last one month.Could some one list what checks I need to do. We would sincerely appreciate the suggestions. Thanks, M.R. PILLAI

1) Check your acrylamide/Bis solutions for any precipitates.2) Make fresh APS and perhaps try another batch of TEMED.3) Make sure that the temperature of the room where the gel is left to polymerize is not too low.4) Double check your protocol and make sure that you are indeed adding the right amounts of the ingredients.

I am guessing you have run enough gels to trouble-shoot the problem and that nothing has worked so far. What Dimitris suggests is very relevant but if you feel that is not the problem, check the filter of your water source- heavy metal sulfates inhibit polymerization. If I am right, adding 1 mM Zn2+ will fix the problem for now, but for long term, make sure the water is clean.

Meghna Vasudevanhttp://www.dstf.bigstep.com

1) Check your acrylamide/Bis solutions for any precipitates.2) Make fresh APS and perhaps try another batch of TEMED.3) Make sure that the temperature of the room where the gel is left to polymerize is not too low.4) Double check your protocol and make sure that you are indeed adding the right amounts of the ingredients.

Also not that the APS is only good for 7 days by the 8th day (you're pushing it) it could or could not work