in vitro transcription

Matt Thomas Thomas at molbio.uoregon.edu
Mon May 19 03:34:57 EST 1997


I don't know about the comercial extracts, but we do a lot of in vitro
txn with Nam cell extracts and in the past Hela extracts. There was alot
of variation between differenct prepes.  I think it is just a fact of
life withuseing this kind of material.  We see this variation in G-less
assays and on more traditional templates.  The only thing I have noticed
is that the extracts that typically give good signal are high in protein
concentration, anything from 4.0 to 8.0 mg/ml are usually the best.  But
this is not a garuentee.  Every now and then I'll get a stinker extract
which according to the protein concentration should be fine.  Maybe you
should try a different supplier.  Just a suggestion.  But making your
own is not a gaurantee it will work and it is not trival to set up large
scale cell culture to produce the material needed.

______________________________________________________________________
Matthew Thomas,
Institute of Molecular Biology, University of Oregon
Thomas at molbio.uoregon.edu

Science is a cross dressing dominatrix.
--a fellow graduate student
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