HELP nuclear runon nuclear runoff
Rick Nicholson
R.Nicholson at cfi.unsw.edu.au
Tue Aug 27 22:08:19 EST 1996
Anis Limami wrote > I would like that somebody explain me the difference
between
> the two methods Nuclear run-off and nuclear run-on.
> I looked in recipe books (current methods in M. B.) I found
> only runoff.
Historically nuclear run-on referred to the extension in-vitro of
transcripts initiated in-vivo. That is the nuclei are isolated and the
RNA molecules which had begun to be synthesised in-vivo are extended, or
run-on, in-vitro.
The term run-off transcription (coined by Kreig and Melton around 1984
in NAR, and Meth. Enzymol 1988) referred to in-vitro transcription from
linearized DNA template. Hence the term run-off, as the polymerase ran
until it fell off the template. (The traditional run-off procedures are
described in manuals such as the Promega Protocols and Applications
guide.)
Somehow during the past decade many people have started using the term
nuclear run-off rather than run-on. Although run-on is a more accurate
term, it seems to have become acceptable to also call it nuclear
run-off.
--
Richard C. Nicholson, BSc PhD
Senior Research Fellow
Centre for Immunology
St. Vincent's Hospital
Sydney, Australia 2010
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