transient transfection assays: Measuring Luciferase activity

[Darren Tyson]

Wolfgang Schechinger wrote:

> Hi you,
> in transient transfection assays ( Ca3(PO4)2 oder DEAE-Dextran) I
> always get a =DF-Galactosidase activity of the cotransfected vector
> (CMV-promoter), but it is not possible to measure any Luciferase
> activity of the pGL3-construct or pGL3 control (Promega). The assay
> system is o.k. (measuring the enzym luciferase obtained from Promega)
> Have anyone any idea???
>
> Cora

Cora,

It could be a problem with your lysis buffer.  Did you add the positive
control
luciferase enzyme to the lysis buffer before assaying?  The pGL3-Control
vector
has always given me very high RLU counts.

Darren