Re: in situ digest

Axel Ducret (axel_ducret@merck.com)
Tue, 18 Aug 1998 11:19:32 -0400

Dear John,

I would strongly recommend that you use a continuous %T gel for the reason that
you mentioned: pore size. I had much better result in doing in-gel digest with
such a system than with a gradient gel. Use a 12 %T gel with a 2x Laemmli buffer

running at 16 mA constant per gel (mini-gel system, 1 mm thickness), the run
will last approx. 2-2.5 h (the key, here, is to run the gel slowly.) With a
minigel, you should get separation down to 6 kDa (running front of Bromophenol
Blue).

Good luck

Axel

--
Axel Ducret, Ph.D.
Senior Research Biologist
Merck-Frosst Canada Inc.
Dept. Biochemistry and Molecular Biology
P.O. Box 1005
Pointe-Claire-Dorval PQ H9R 4P8
Canada

tel. + (514) 428-3428 fax + (514) 428-4900