Pouring sequencing gels w/o leaking

Dennis J. Templeton djt2 at po.CWRU.Edu
Tue Jul 6 14:11:05 EST 1993


In a previous article, Miao at oregon.uoregon.edu (Vivian Miao) says:

>In article <21a41p$den at news.bu.edu>, pfoster at bu.edu (Patricia Foster)
>wrote:
>> 
>> James Gibbs (gibbs at husc4.harvard.edu) wrote:
>> 
>> : The only problem I have with pouring horizontal gels, letting capillary
>> : action fill the plates, is bubbles. If bubbles occur, how can you get them
>> : out?
>> 
>> (stuff deleted)
>> 
>
>Ok, I can't stand it; I want to get into this fray of home remedies too!
>
>Does anyone else "fish out" the bubbles?  I'm not into banging on the glass
>(makes me nervous), but when I do get bubbles, I sort of fish them out
>using a long piece of spacer material that has a hook shape cut at the end
>of it. 
>I extend the  hook (has to be clean, of course) past the bubble, draw it
>back

OK, one more into the fray.  We use both the no tape horizontal appraoch
and the bubble hook.  The best bubble hook, IMHO, is made from photographic
film. Cut a shape that reminds you of Ahab's tool (no not that one!) and
that is long enough to reach the middle of the gel. BUT DON'T USE IT YET! 

Remember that film is covered with silver emulsion in gelatin, and this
layer swells when it wets so that it is more than 0.4mm thick.  You can
easily remove the emulsion layer with NaOH (maybe 100 mM) which strips away
the gelatin leaving a thin plastic sheet that is about 0.25 mM thick.  We
used to make spacers and combs fromthis stuff too, but the modern sharks
tooth combs are better.

Happy Hooking,
dennis



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