sequencing gel won't stick to Whatman paper

Paul N Hengen pnh at kaylor.ncifcrf.gov
Thu Jun 13 10:51:03 EST 1996


jlight at SICKKIDS.ON.CA wrote:

: After I run a sequencing gel (using 35S-dATP for labelling) I have a lot of
: problems getting the gel to stick to Whatman paper.  I don't fix the gel
: after the run, because I've found that it's not necessary to get good
: autorads.  Unfortunately, and perhaps because I skip the fixing step, my
: gels don't always stick when I overlay them with Whatman paper (sometimes
: they do, sometimes they don't).  I've tried spraying the gel with water and
: blotting off the excess with Kim wipes prior overlaying with Whatman, which
: helps sometimes, but it's no guarantee.  Sometimes part of the gel sticks
: and part doesn't, so the gel ends up ripping.  Advice from anyone who
: routinely transfers their gels without fixing would be greatly appreciated.

There is a specific amount of liquid needed on the gel before the gel will
stick. I used to fix the gel and then completely drain off the fixer, then
spend 30-45 minutes trying to carefully peel the gel from the plate with
a dry Whatman paper. I had a huge amount of trouble with this until I talked
to a post-doc in our lab, Vinay Thatte, who was very experienced at this. He
bet me a pizza that he could stick the paper on and peel the gel off in one
hand motion under 10 seconds. I lost a pizza because he actually did it! He
knew exactly how much fixer to leave on the gel before placing the paper on.
I think the person who suggested wetting the paper first might have the answer.
I suggest spraying on water from a windex bottle rather than soaking a sheet of
paper in a puddle.  That way the paper has some sticking power left in it :-)

--
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* Paul N. Hengen, Ph.D.                           /--------------------------/*
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