gel electrophoresis of CNBr fragments

Deb McMillen (mcmillen@morel.uoregon.edu)
Thu, 13 Aug 1998 15:41:34 -0700 (PDT)

Hi, all,
I have a researcher who would like to run gel electrophoresis of CNBr
fragments (ranging from 1000 to 6000 daltons) of a protein. We are
looking at the Tricine-SDS gel electrophoresis method of Shagger and von
Jagow. We normally do our CNBr cleavage in 0.1 M HCl with 30%
acetonitrile. We will want to load the freshly digested sample directly
on the gel--we plan to lyophilize the samples to remove acetonitrile (but
not taking the sample to dryness) and to adjust the pH up to 6.6 for the
sample application to the gels. Are there any other suggestions for
sample handling (or is there a reference for direct application of CNBr
fragments to gels?)

Thanks for your advice,
Deb McMillen
Institute of Molecular Biology
University of Oregon