Fixing/drying silane bound seq gels
Dennis J. Templeton
djt2 at po.CWRU.Edu
Mon Jun 15 15:11:54 EST 1992
In a previous article, 37_527 at uwovax.uwo.ca (Gregory A. Denomme) says:
> A number of postings have been made concerning fixing/drying sequencing
>gel but are not applicable for those who bind the gel to the plate using
> How about some tips or shortcuts for this method?
> I have found that occasionally areas of the gel crack and peel off the
>plate. This occurs near the end of the drying step. I usually dry my plate
>about 1.5 hrs at 65oC. I am not sure why this happens. I have increase the
>fixing time to 1 hr and the soaking time with H2O to 1.5 hrs. I'm thinking
>now that it may be related to the drying temp. Any comments?
We use this method too, and the only time we've had "peeling" problems, it
was due to not having enough "bind" silane in spots. Our recipe:
10 ml EtOH (95%)
100 ul Glacial Acetic acid
15 ul bind silane (from sigma, methoxypropylsomethingsilane
Rub on with kimwipe, let dry, rinse well with 95% Etoh, dry again
We don't fix anywhere near as long as you, maybe 15 minutes in regular gel
fix with agitation. I'd be afraid of losing signal from small-sized bands
with a long soak. Then we rinse for just 2 mins in water
we dry at RT with a fan, since we don't have a big oven.
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