sequencing gel won't stick to Whatman paper

Jasper Saris jasper at ruly46.medfac.leidenuniv.nl
Tue Jun 18 07:43:57 EST 1996


Paul N Hengen (pnh at kaylor.ncifcrf.gov) wrote:
: 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.

Hoi,

For several years now we don't fix the sequencing gels. Both plates were
repelled (Pharmacia) and we used Whatman 3MM. For several months (>6 ) we
have changed to S&S type 2 paper and Pharmacia changed their Repel to
Repel ES (Extra Strong?). Some three months ago I also have had
difficulties with gels sticking to the glassplates instead to the S&S
paper, but mainly with 10% PAA 1/50 + Glycerol SSCP gels.

The repel has certainly effect during pouring, but I don't know for sure
if it one of the variables in the sticking problem. The front, larger
plate is now treaten less often than the smaller, back plate. Saves again
time, but beware. 
 I treat the paper a bit different now. Longer lying on the plate was not
the help I hoped for. An over dessicated surface could be, but I proceed
immediatly after dismounting the two plates. Pressing the back of the
paper with the back of a razor blade did help better than waiting for
several minutes (1-10min). Not flipping the paper-gel-plate set with the
second plate, but just lifting the paper over itself worked the best for
several gel types. 

Good luck with thread of advice, it is a lot to choose from


Jasper

J.J. Saris
Dept. Human Genetics
Leiden University
Sylvius laboratory
wassenaarseweg 72
2333 Al Leiden





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