hekp with sigmacoat and sequencing gels

Thomas Chuang tommyc at caa32.alumni.berkeley.edu
Sun Jul 23 13:24:29 EST 1995


Jeffrey Mahr (szjmahr at peseta.ucdavis.edu) wrote:
: I am relatively new to sequencing.  at first when pouring my gels i would 
: clean them with detergent and a sponge, lay them flat on a tt rack and 
: with no tape, pour the gel - this worked great until once when taking the 
: apparatus apart the gel stuck to both plates and made a mess of it.  to 
: try to prevent this i tried using sigmacoat on one plate and disaster 
: struck (huge pockets where the acrylamide just WOULD NOT go - the 
: location of which where not the same when i tried this 3 times in a 
: row).  for my fourth attempt icleaned the plates using every dish washing 
: solvent in the lab and did not use sigma coat - the same thing to a 
: somewhat lesser degree happened again (i even tried taping halfway 
: through and then bringing the plates vertical but the huge bubbles 
: wouldn't budge despite my pouding on the plates)  i have heard mixed 
: reveiws on sigma coat
: 	1.  will i ever be able to get the stuff off my plates
: 	2.  does anyone know of a way to use the stuff so that it works reliably
: 	3  does any one have  alternative ideas for helping me out????

After another round of sequencing, I just use warm water and some ALCONOX 
detergent to let sit the plates in a tub. When ready for use, scrub a 
bit, rinse and dry the plates. Apply RAIN-X on both sides of the smaller 
plate of the plate sandwich (or the one plate that you are lifting to leave the 
polyacrylamide gel on). Rinse and dry the plates again. Use 95% EtOH wash 
and wipe dry with large Kimwipes. Then you're ready to go. Once RAIN-X is 
applied, I found that it lasts 5-6 times of sequencing until 
re-application is necessary.

Hope that helps,


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			    University of California at Berkeley
      	Tom Chuang		-= Home of Golden Bears =-
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