FROZEN acrylamide sol'n!

Duke Groebe drg at prophet.pharm.pitt.edu
Thu Apr 18 09:51:22 EST 1996


In article <4l2t3c$17l at ccshst05.uoguelph.ca>, tjackson at uoguelph.ca (Timothy
R Jackson) wrote:
> 
> we are making our own 8% denaturing acrylamide gel solution for 
> electrophoresis, sequencing, etc. and were told by a few people that it's 
> best to store the stuff in a 4 degree fridge.  well our damn acrylamide 
> solution freezes!!  some days it doesn't some batches don't - it's a real 
> pain!  however, since we started doing this, when the acrylamide isn't 
> frozen, we're getting the nicest gels we've ever run.  what's the scoop 
> here?  why does it freeze and what can we do to prevent it?  any help 
> here would be greatly appreciated.  if it's any help, we're making 7.83M 
> urea gels and mixing the solution in a vacuum (i.e. degassing).
> thanx in advance!


If you have urea in your gel stock, leave it out.  *Everyone* knows that
you're supposed to make urea solutions fresh anyway because the stuff
breaks down.  Make up a higher concentration of stock acrylamide solution
(I think 40% is recommended) in H2O and dilute to make your final gel
solution.  So, for 50 ml of gel stock at 7M urea and 8% acrylamide, you'll
need 21 g of urea, 5 ml of 10X TBE, 10 ml of 40% acrylamide solution (which
is actually 38% acrylamide, 2% bis-acrylamide), and about 19.3 ml H2O.  Mix
under vacuum.  Redi-2-use.

Duke

******************************************************************************
The opinions expressed here are my own.  MY very own and NOBODY else's!  I
THOUGHT OF THEM FIRST, HA HA, AND YOU DIDN'T!!  SEE!?  There's my name up
there!  And the date - Ooohh, the DATE!  That's important!  HEE HEE, I'M
GONNA BE RICH!  DO YOU HEAR ME!?  RICH!!  HA HA!!  They're mine...heh, heh,
all mine....  GOD!  I'm so smart!



More information about the Methods mailing list