labelling in gel slice

bugg at mbcf.stjude.org bugg at mbcf.stjude.org
Mon Jun 28 10:07:55 EST 1993


Hello, netters!
Joel L. Vanneste of The Horticulture and Food Research Institute of New
Zealand, Ltd. has requested that I post the reference for the "followup" paper
of Feinberg and Vogelstein on this newsgroup.  In this reference, the authors
elaborate on their earlier method to include labelling restriction fragments in
the agarose gel without any purification.  Since I have used this method ever
since it was published with great success, I suggested it to someone on this
newsgroup, but I must have forgotten to give the reference.  It is:
A.P. Feinberg and B. Vogelstein "Addendum: A technique for radiolabelling DNA
restriction fragments to high specific activity" Analytical Biochemistry
137:266-267 (1984).  They suggest using BioRad low-gelling temp agarose, I use
FMC's SeaPlaque GTG, but it probably makes no difference as long as you use
very pure low-gelling temp agarose.  I have a friend who prepares lots of
fragment for labelling at once by running a preparative-well gel, slicing it
out, melting it, and then aliquoting volumes which contain approx. 50-100 ng of
fragment into separate sterile tubes.  She puts the tubes in the freezer and
boils them for 5 min whenever she needs to label that fragment - then adds the
rest of the stuff for the reaction.
Best wishes, everyone!

(By the way, I know this is not the group to mention this, but there is a
postdoctoral position open in our lab and I have posted the opening on the
Bionet Jobs.  If anyone would like more information, please e-mail me
directly.)

Barbara Bugg
Molecular Pharmacology
St. Jude Children's Research Hospital
Memphis, TN  USA



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