Sequencing Gels Not Sticking to Whatmann

Bryan L. Ford fordb at bcc.orst.edu
Mon Nov 10 01:15:52 EST 1997


Rob Kirkpatrick wrote:
> 
>    One of the people in our lab is having a unique problem.  After running
> her sequencing gels she places the Whatman paper on the gel to remove it
> from the front plate and then disaster strikes.  The gel does not want to
> come off the glass plate and rips to shreds.  This has happened twice now
> and I can't figure it out.  The backplate is "Sigma Coated" so we can't
> really apply this to the front plate (or can we?).  We haven't changed the
> Whatman or anything else that I can think of...  Has anyone ever
> encountered this before???


Rob:

I concur with most of the other suggestions your inquiry has elicited.
Recently I noticed one novel source of excessive stickiness: if one has
weak or old ammonium persulfate in the mix there is a slower
polymerization of the gel, this appears to result in greater adhesion to
glass surfaces even if they are silanized and quite clean. My surmise
from this experience is that normal rapid polymerization tends to leave
a thin layer of excluded buffer between the gel and the glass, whereas
slow polymerization results in polymer right up to intimate contact with
the glass surface. Such slow-polymerized gels are very difficult and
sometimes impossible to remove, without tearing, from the glass
plate(s).

So while you're experimenting, I would recommend you try say 50% more of
your "old" ammonium persulfate-- And to give you time to pour/fill the
gel in case this is way too fast, keep your mix on ice right up to the
pouring/filling time.

Good luck,

Bryan




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