Protein purification after in vitro transcription/translation using Roche RTS100
junk at hgmp.mrc.ac.uk
Thu Nov 28 09:01:57 EST 2002
Historians believe that in newspost
<Pine.SOL.4.33.0211261734040.2229-100000 at mcmail.cis.mcmaster.ca> on Tue,
26 Nov 2002, Kyle Legate <legatek at mcmail.cis.mcmaster.ca> penned the
following literary masterpiece:
>This sounds like an incredibly expensive approach. In my experience the
>amount of protein made in an in vitro retic system is in the neighbourhood
>of 1-5 fmole/microL using an SP6 promoter.
Not really. The Roche system, specifically how it works, increases yield
to hundreds ug/ml if not the odd mg/ml of final protein. You can scale
to 10's of mls (or whatever it is).
In most labs the expensive bit is time. Cloning to get the odd mg of
active protein, especially something that is lethal, could take a lot
longer and therefore cost more $ than using the Roche system and with
the latter you have the protein in under a week.
I am not a user but can see benefits in Roche's approach if you have the
I love deadlines. I especially like the whooshing noise they make as
they go flying by.
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