Sequencing gel combs (leakage)

Paul N Hengen pnh at fcs260c2.ncifcrf.gov
Wed Apr 6 12:08:16 EST 1994


 In article <CnGD5B.KD4 at cc.umontreal.ca>
 coady at ERE.UMontreal.CA (Michael Coady) writes:

> Despite all efforts to make my plates perfectly clean, and
> despite using a silicone treatment, I still get little bubbles.  I
> have been pouring plates that are perfectly horizontal; in fact,
> the word pouring isn't really applicable to the capillary process
> that's involved.  In any case, I got rid of my bubbles by including
> a very thin drawn quartz tubing in the gel, at one side.  After the
> gel was poured, I held the tubing at both ends and carefully drew
> it across the gel.  It pulled the bubbles to one side pretty well.

> I didn't need to use all the wells, so I was happy enough to get 
> all the bubbles to one side.  If need be, I could have made a big
> loop with this tubing and pulled it right through the gel, bringing
> the bubbles along with it.  The tubing is available down to 
> 0.165 mm O.D., so it can be used for 0.2 mm gels.  If anybody wants
> to try it out, I can dig up the retailer.  I've gotten it from a 
> lab that's used it for renal tubule perfusion, and for use in
> electrophysiology.
>
> Mike

I wonder if you could use something a little less expensive like a heavy test
fishing line. These come in all diameters so that you might be able to find one
just the size you need to fit between the plates. Also, some of these come
tapered -> for example, those used for the ends of fly fishing lines.  One of
these could be used for removing bubbles by inserting the thin part first, then
trailing it through with increasing thickness coming last to snag the little
bubblets. I've never tried this, but I just thought about it. It sure beats
tapping the glass with a pair of sissors or screwdriver handle as I've seen
many people do. My feeling is that tapping leads to microscopic nicks in the
glass plates which then act as nucleation sites or hanging spots for bubbles
the next time you pour the gel _AND_ I hate the way that sounds when I'm
trying to concentrate on something. It drives me nuts.

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* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
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