Problems with 377 48cm gels

Macky Edmundson mackyE at qimr.edu.au
Thu Jun 13 04:29:53 EST 1996


Hi again,

thanks for the replies from everyone, It sounds like I will just have 
tolive with loose combs on 48cm gels.
Our problem with leaking upper buffer is not usually due to leaking of 
the seals but seems to be due to wicking of the buffer up and over the 
spacers.
I was overfilling my early gels and the wicking was a lot worse but 
even now  I am still  having problems. Squirting 3% agarose over the 
spacers seems to help but it is a pain to have to do it for every run.

The acrylamide is polymerised right to the top though there is always 
a small bubble formed in the "v" of the spacer. Wicking does not seem 
to be related to the size or number of bubbles anyway.

Margret Keller writes ;

> We have found that the 377 pouring apparatus is cumbersome to use,
>especially with the longer plates. We are currently using the Sooner
>Scientific Gel Caster, with no complaints. As far as the leakage problem, >have you considered that your pouring problems may lead to less acrylamide >up around the comb corners contributing to your leakage problem?
>
>We have a 377 problem as well. Both with 36 and 48 cm plates, we get very noisy plate checks that do not clean at all. Plates are washed with water only. The 48 cm plate set was never used before, so I cant believe it is a
 build-up problem. We have loaded around the noisy lanes and these 
noisy lanes show up as green streaks on the gel image. This makes it 
currently impossible to get a full gel on our 377 and is very 
frustrating. Any suggestions would be appreciated!
>Margaret Keller

I has taken me about a month to perfect gel poring with the ABI 
apparatus and while I still miss the simplicity of slide pouring of 
our old 373 gels it is working O.K. for now. The most important thing 
with the 377 plates is to wash the plates with very hot water, then 
allconox , scrub with wet kimwipes, then hot water again, and finally 
with deionised. About once a month squirt the plates with saturated 
KOH in ethanol , let it dry for a few minutes then wash off. Just 
before gel pouring put some ethanol on 2 kimwipes and scrub the inside 
of the plates VERY hard and as fast as possible. If I do not do this I 
cannot get the gel to pour properly.

If I have problems with an area of the plates which are not wetting
properly (like the area around the seal) I scrub the area with
dimethylformamide (acetone should work as well) on some kimwipes , 
then wash normally and that fixes it.

I find that 377 is much less sensitive to gel background problems than 
the 373 now that I have got the gel pouring sorted out.

Tip of the Day

To help protect your gel plates go to any carpet dealer and buy some
non-slip backing for rugs. It looks really weird as it is a grid of  
soft rubber blobs stuck together. We use two layers on the drying
bench and in the sink and it works much better than the shower mats 
some people use as it allows the plates to dry and it is cheap.


   Macky Edmundson - Queensland Institute of Medical Research

        Socrates         - To Be is To Do
        Jean Paul Sartre - To Do is To Be
        Frank Sinatra    - Do Be Do Be Do

        email mackyE at qimr.edu.au  (Australia)
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