SSCP detection

HARDIES at THORIN.UTHSCSA.EDU HARDIES at THORIN.UTHSCSA.EDU
Thu Dec 8 14:59:17 EST 1994


Jim Schupp writes:

>        We have been using AT Biochem's MDE gel for SSCP detection and
> always have a hell of a time transfering the 0.4mm 14X17 gel to filter
> paper for drying and film exposure.  The gel is very sticky, even on
> rain-x treated glass plates.  More often than not, we get distorted gels
> due to the gel stretching during transfer.  Anyone have similar experiences?
> Should we not be transfering to filter paper, just exposing the gel while
> it is still attached to the glass?

I just take off one plate, put the other plate with the gel in a 
plastic bag, and expose direct.  Since I use P32, it never occurred
to me that there would be any reason to dry the gel; is there?

When you say you have trouble transferring to filter paper, are you
laying a dry Whatman sheet on the gel and then peeling it off like 
for a sequencing gel?  I never used MDE, but it sounds like you're
saying it sticks to the glass tighter than the paper.  I have seen
people have this problem with polyacrylamide because they let the
paper get too wet.  Is part of the problem the large surface you're
trying to transfer?  Maybe you cound find the counts with a Geigor 
counter first, lay down a smaller sheet of paper, and then cut around
it.  The smaller paper might be easier to lift off.

Hope this helps.
Steve Hardies, Dept. of Biochem., Univ. of Texas HSC at San Antonio
Hardies at thorin.uthscsa.edu




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