hekp with sigmacoat and sequencing gels

Youre-mailaddress Youre-mailaddress
Wed Jul 12 15:37:49 EST 1995


In article <Pine.SOL.3.91.950711134256.15100A-100000 at chip>, 
szjmahr at peseta.ucdavis.edu says...
>
>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????
>
>
>
>sincerly - jamahr at ucdavis.edu
>

I use SigmaCote straight from the bottle, using a disposable pipettor, 
(perhaps a total of 6 drops/plate) and distributing evenly with a 
Kimwipe.  I let the plates air dry.  I routinely coat both plates, then 
hold the sandwich at about a 30 degree angle and pipet the acrylamide 
solution in with a disposable transfer pipet into the top corner, letting 
capillary action and gravity fill the plates.  Sometimes it takes awhile 
to fill, sometimes it fills very rapidly.  A BubbleBuster (AT Biochem) is 
the best way to get rid of bubbles--and to use to start the filling if 
the SigmaCote is really working.  I use a 2% NaOH solution to get rid of 
the SigmaCote--although normally I just wash and reuse my plates.  The 
SigmaCote usually lasts 3 plus times before I have sticking problems 
again.  I prefer SigmaCote to Gel Slick (AT Biochem) because I prefer to 
have a slow-pouring gel than one that sticks, tears, and ripples when 
transferring to filter paper.  Good luck.




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