Problems with SDS-Page

Thu Apr 21 09:29:55 EST 1994

In my experience, misshapen wells in the stacking gel of SDS-PAGE are 
caused when the well-forming comb is removed 'late'.  For some reason 4% 
acrylamide, when fully polymerized, tends to stick to the teflon comb, so 
wells become distorted upon comb removal.  We get perfectly well formed 
wells (pun intended) when we remove combs from the stacking gel at about 15 
minutes after TEMED addition (room temperature).  There will still be some 
liquid that remains in the wells, but we simply blot it out with Whatman 
3MM strips; if you leave the fluid in the wells, it will polymerize.  After 
blotting, the wells have sharp sides and corners and are of uniform size.
Hope this help!  Cheers,  Shaun
  = Shaun D. Black, PhD   | Internet address:     shaun at = 
  = Dept. of Biochemistry | University of Texas Health Center, at Tyler = 

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