I Used 30% acrylamide, separating buffer Ph-8.8, 10%SDS, 20%APS and TEMED to prepare 10% acrylamide, but it turns cloudy white after solidification. Can anyone tell the reason behind it?
Was 10% SDS the final concentration of your separation buffer or was that a stock solution and you've taken out a portion (like 100 ul of 10% SDS into 10 ml final volume of buffer) to make the separation buffer? Judging by that one photo it looks like the SDS has crashed out of solution
Try using the attached protocol. Gels prepared this way never caused any problems for me.
http://cshprotocols.cshlp.org/content/2015/7/pdb.rec087908.full?rss=1
Dear Albert Lee, thanks for the reply; I also thought the same, so the second time I did not use SDS, but the same outcome was there.
If it is not the SDS, then that leaves APS/TEMED and the acrylamide. But if the gel is solidifying you can assume APS and TEMED are working fine. Acrylamide/Bis-acrylamide are light and air sensitive. Try to use a different batch of these chemicals as the one you have may have expired/spoiled. I don't think this is a pH issue unless the pH of the Tris you are using is way off.
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