Protein purification after in vitro transcription/translation using Roche RTS100

Duncan Clark 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 
initial $.

Duncan
-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.



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