seqn gel pouring
halligan at post.its.mcw.edu
Fri Jul 9 11:58:11 EST 1993
The method that I learned as a post-doc and we still use in my lab is to
place a strip of Whattman No. 1 filter paper (1 cm wide tape) at the bottom
of the plates in place of a bottom spacer. We then pour horizontally with
no tape, but clamps around the edge. The paper spacer seems to let air pass
(it is slightly thinner than the spacer material) but the gel mix causes it
to swell and form a loose seal. The advantage of this method is that the
paper tape is porous and does not have to be removed before running the gel
(this also avoids bubbles at the bottom of the gel).
Don Back (BACKD at QUCDN.QueensU.CA) wrote:
: In article <20tgrp$94v at usenet.INS.CWRU.Edu> bl275 at cleveland.Freenet.Edu (Dan Diaz) writes:
: >From: bl275 at cleveland.Freenet.Edu (Dan Diaz)
: >Subject: Re: seqn gel pouring
: >Date: 1 Jul 1993 02:06:49 GMT
: >In a previous article, brett at BORCIM.WUSTL.EDU () says:
: >>to *eliminate* gel leaking simply do this: catalyze ~5 mls of your gel soln
: >>and pour into your gel from the top, keeping yuor plates VERTICAL. Use enough
: >>TEMED for it to polymerize in about 10-20 min. When set just pour the rest on
: >>top, lay flat and insert comb of your choice. Because you pour it vertically,
: >>all bubbles will rise to the top and pop. I might add that I pour the 5 mls
: >>onto loosely taped plates without any leakage. The 5 mls is strong enough to hold
: >>the gel above it.
: >my lab doesnt bother with tape or acrylamide plugs or nuttin. Clamp your
: >plates on the sides. Prepare your acrylamide solution and lay your plates
: >flat on a few empty pipet tip boxes over some disposable bench matting
: >material. Put your acryl mix in a plastic 100 mL cylinder and begin slowly
: >pouring near one edge of the plates. Move the cylinder slowly across to
: >the other side as you continue pouring slowly. When you have gotten to the
: >other side and the acrylamide front is progressing down the plates, return
: >the cylinder to the center and continue pouring slowly. Capillary action
: >will do the rest. If your plates are clean, bubbles will be rare, and
: >should they occur, can be removed quickly with a long bent piece of thin
: >wire. When the acryl solution reaches the bottom, a bit will spill, but
: >thats what the bench matting is for. Insert the comb, clamp, voila. A tad
: >messy, but it's easy and its yet to fail us.
: >Dan `Diz' Diaz "My opinions are those of all intelligent people"
: >BL275 at cleveland.freenet.edu
: I stopped taping plates years ago. My technique is a bit different from
: Lay the bottom plate level on a couple of pipet tip boxes, just to get it up
: off the bench. Place a layer of absorbent bench paper down first.
: Set the spacers on the bottom plate so that only the top part of the spacer
: is on the plate and the rest is dangling onto the bench.
: Pour the acrylamide solution more or less evenly over the bottom plate.
: Make certain that you have one continuous bead along the bottom of the plate.
: Set the top edge of the top plate along the bottom edge of the bottom plate.
: Slowly slide the top plate along the bottom plate. This will draw the
: spacers up along with the top plate.
: The acrylamide will be drawn in by capillary action as the plates slide
: along. Watch the bottom edge, if you slide the plate too quickly you will
: draw air into the bottom.
: When the plates are in position insert the comb and clamp. The excess
: acrylamide will leak out onto the bench paper.
: I find the surface tension formed along the top edge interface as you slide
: the top plate up traps any bubbles that might form. The advantage is that
: you don't require capillary action to draw acrylamide all the way to the
: bottom of the gel.
: Give it a try, it sure cuts down on preparation time.
: Don Back
: Department of Biochemistry
: Queen's University
: Kingston, Ontario
Brian D. Halligan halligan at post.its.mcw.edu
Assistant Professor 414-257-8413 (voice)
Department of Microbiology 414-257-8427 (FAX)
Medical College of Wisconsin
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