SUMMARY Sequencing gel combs (leakage)

the End jgraham at bronze.ucs.indiana.edu
Thu Mar 31 13:31:42 EST 1994


Fellow Bionetters,

Thanks everyone for the hearty response. Another demonstration of
the power of the traditional free exchange of information unhampered
by patents, profiteering, and proprietary materials. :)

After clamping the *&$#@ out of the top of my plates during polymerization
while leaving the edges unclamped (as Mike suggests) I've seen vast
improvment.

Following an editied repost of Mike's good word is a nice addition sent to me by Michael Finney that I intend to try shortly.

Jim
J. Graham

_____________________________

               
From: tencza at med.unc.edu (Michael G. Tencza)         29 Mar 1994 14:42
Organization: UNC-CH School of Medicine

The first recommendation I can offer is to make sure
the best combination of long plate to bottom plate is achieved, thus
optimizing the spacing between plates where the comb fits in.  The next thing
is to make sure your comb is replaced in the same orientation between the
plates when you remove the flat side and replace the tooth side (ie: don't
flip the comb right to left).  The last and possibly most important tip is
to place the top clamps on the glass directly over the comb and not the side
spacers during polymerization of your gel.  This normally insures a good
seal and little or no leakage will occur between lanes.

Good luck,

Mike

_______________________________

From: mikef at mjr.com (Michael Finney)
Organization: MJ Research, Inc.

I read the first 7 posts to follow up your question, and I second the
consensus about clamping the plates while polymerizing.

But there are further tricks to make the gel polymerize solidly right up to
the comb.  These seem to make a real difference.
1  make sure the gel mix is very thoroughly degassed.
2  wipe the flat edge of the comb with a kimwipe soaked in 10% APS before
inserting it between the plates.

The second rather clever trick can be traced back at least to Nancy
Hopkins's
lab at MIT, but may not have originated there.

Mike

.




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