TNT kit with T7 RNA Pol

Kojo Mensa-Wilmot mensawil at
Sat Mar 30 10:17:53 EST 1996

Hans Bernd Prisack wrote:
> In article <314E1D33.662A at> u47888 at UIC.EDU (HONG) writes:
> >From: u47888 at UIC.EDU (HONG)
> >Subject: TNT kit with T7 RNA Pol
> >Date: 18 Mar 1996 18:36:45 -0800
> >Has anyone figured out how to get consistent results with T7 RNA pol.
> >using promega TNT kit?  We get good results with sp6 RNA pol, but not
> >with T7 RNA pol.  DNA templates used in T7 were linearized according
> >to the promega protocol.
> >Thanks for any input.
> >Hong
> >UIC
> I am also trying to get good translation products using the Promega TNT kit
> with T7-Polymerase, but until yet I didn´t. So I would be very appriciate to
> get any suggestions for this problem. I´m trying to translate PCR-products
> which contain the T7-RNA-promotor and label the protein with biotin-tRNA. I
> got a protein with the control-luciferase-vector but not with my PCR-products.
> Prof. Dr. med. H. Bojar
> Institut fuer Onkologische Chemie
> Gebaeude 23.12 Ebene U1
> Medizinische Einrichtungen
> Heinrich-Heine-Universitaet
> Postfach 10 10 07
> 40001 Duesseldorf
> Tel.: 0211/311-4322
> Fax : 0211/311-5114
> E-Mail:bojar at
We have had consistent using TNT on supercoiled 
plasmids. An attempt or two on linearised plasmid failed. 
This might indicate that your pcr product needs to be 
cloned into a vector before using TNT for analysis.

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