TNT kit with T7 RNA Pol
mensawil at cb.uga.edu
Sat Mar 30 10:17:53 EST 1996
Hans Bernd Prisack wrote:
> In article <314E1D33.662A at uic.edu> 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.
> 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
> Postfach 10 10 07
> 40001 Duesseldorf
> Tel.: 0211/311-4322
> Fax : 0211/311-5114
> E-Mail:bojar at uni-duesseldorf.de
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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