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:

>Hello Netters;
>  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
>silane.
>
>  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?
>
>-Greg

Greg,  

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.

Good luck, 
dennis



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