Bind dilane protocols for seq gels

Bill Melchior, NCTR/FDA wmelchior at NTDOC.NCTNET.GOV
Tue Feb 25 15:23:22 EST 1992


When treating the plate the gel is to bind to, it is important to 
COMPLETELY remove excess chemical.  I finish by spraying the plate with
95% EtOH, wiping thouroughly with tissue, and wiping dry.  I repeat this
THREE times, drying each time.  (I have read that the chemical can 
diffuse through the gel and cause binding to the other plate.  Some 
workers here who were not properly wiping the plates before pouring their
gels did in fact have the gel stick to both plates.)

If the BDH compound is gamma-methacryloxypropyltrimthoxy silane (phew!),
I have good luck with the following:

10 ml 95 %EtOH to a disposable testtube.
Add, in order, 50 microliters silane and 30 microliters glacial HOAc.
Vortex briefly.  Pour half the solution on the previously cleaned plate,
  wiping very well with a tissue.  (Wear gloves.)  Allow to dry to a haze 
  (a few minutes).
Pour the rest of the solution on, again wipe well, and allow to dry for a
  few minutes.
Three times:  spray with 95% EtOH and wipe well until dry.


I used to use a chloroform or heptane solution of dichlorodimethylsilane
to treat the other plate, but it's easier and safer to use Rain-X from 
the auto department of my local discount store, used according to the
bottle instructions.

Bill Melchior                                ||   Evolution, as described in
National Center for Toxicological Research   ||   J. A. Paulos' _Innumeracy_:
Jefferson, AR  72079                         ||
(501) 543-7206                               ||  "Eventually, primitive life
                                             ||   develops, and then shopping
wmelchior at ntdoc.nctnet.gov                   ||   malls."



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