TNT in vitro transcription/translation system

user at mpi.com user at mpi.com
Sat Oct 11 19:51:34 EST 1997


I used to do that routinely and it worked fine.  It's been a few years, so
I don't remember how many units I used...I think the same units for T3 and
T7. Anyway, you have nothing to lose by trying it - at worst you waste 1
or 2 reactions worth of reagents.  (I remember wondering whether the two
systems were really different at all, or just a way to get people to buy
more stuff...)
Hilde



In article <fof1-1010971243410001 at zellkultur.biologie.uni-konstanz.de>,
fof1 at chclu.chemie.uni-konstanz.de (Frank O. Fackelmayer) wrote:

> Hi all,
> We sometimes use in vitro translation to get protein synthesized from cDNA
> clones. In the last time, we´ve used the Promega TNT system for coupled in
> vitro  transcription/translation, which works very well. We have been
> using the T7-polymerase based system, and now I need a T3-polymerase
> system for only one or two reactions. I got plenty of the T7 system left,
> and I thought that adding T3 polymerase instead of the T7 polymerase
> should do the job (reaction conditions for the two polymerases are
> essentially the same in standard in vitro transcription assays). Has
> anybody tried that? Successfully? How many units of polymerase do you use?
> I really dont want to buy a complete T3 system for only one or two reactions.
> 
> Thanks,
> Frank



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