Pouring sequencing gels the old fashion way
colin at fungus.biochem.ualberta.ca
Wed Apr 17 21:04:45 EST 1996
Rob Jordan wrote:
> I know that everybody these days is raving about new, fast ways to pour
> sequencing gels, but let's assume for the time being that I want to do it the
> old fashioned way and tape up the edges of the gel. I've tried three types of
> tape since moving to a new lab and all of them end up leaking (badly). Can
> anyone give me a brand name and/or taping method they use to pour leak-proof
> gels? Every time I've used a gel that has leaked I end up with lanes sucked
> to one side or the other.
Having sequenced since the good old days I would recommend the following. Forget
the tape. Clamp your gels together with binder clips (with the spacers in there
and all). Take about 10 ml of gel mix and add 5 times the normal APS, and then 5
times the normal TEMED. Working quickly (you have about 15 sec), apply with a
P1000 to the bottom of the gel to create a plug. Let set, then pour the remainder
of the gel using the normal amount of APS and TEMED. Using 3 or 4 clamps per side
creates enough pressure that the gel won't leak if poured at a shallow angle and
then let stand flat while the gel polymerizes.
Alternatively, get one of those gel casting boots from BRL. They work great, no
tape, no plugs, no clamps, just pour the gel. I got one recently and I will never
pour sequencing gels any other way.
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