in vitro transcription

Matt Thomas Thomas at
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

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