luciferase

daemon at net.bio.net daemon at net.bio.net
Sun Feb 9 20:25:16 EST 1997


Dear Dr. Planchais,

     There is some literature saying you can use scintillation counters,
especially if you switch off their coincidence circuit.  However, I found
that if you do this, the background is horrible, and I just wasted my time
fooling with it.  I recommend a real luminometer.

     I expect GUS and luciferase are stable in the same buffer, although
you don't tell me the buffer.  Just be sure that you add 10 mM DTT fresh fro=
m
a stock of 1 M DTT stored frozen in water.  Or mercaptoethanol.  Last I chec=
ked,
I had not seen the importance of reducing agent in any literature about
luciferase, except in my 1990 PNAS paper, but I found then that it is critic=
al
to reproducible luciferase assays.  I was able to bring year-old Sigma
luciferase
back to life with this DTT.


Wayne M. Barnes, Ph.D.                        wayne at barnes1.wustl.edu
Biochemistry Dept. 8231                   or  barnes at biodec.wustl.edu
Washington Univ. Medical School               314.362.3351  fax 7183
660 South Euclid Ave., St. Louis, MO 63110
           http://mbb.wustl.edu/~barnes/

> Dear Pr.Barnes,
>
> You sent me few months ago the pWB216 construct containing the 35S promote=
r
> upstream the luciferase gene. I have some questions about the luciferase
> assay. I would like to know if it is possible to determine the quantity of
> light with a scintillation counter. Is it possible to extract GUS and
> luciferase activity in the same buffer? Did you use the luciferase assay
> system from Promega?
> Hoping to hear from you soon,
> Best wishes
>
> Severine Planchais
>
> Institut de Biotechnologie des Plantes
> Universit=E9 Paris-Sud
> Batiment 630
> 91405 ORSAY cedex
>







More information about the Arab-gen mailing list