1 reply to this topic

[Pkrish]

Hi

I am performing luciferase assay with 2 expression vectors+ 1 promoter-luciferase vector+GFP- control vector.

So each time i perform the experiment I get extremely different readings.

Could it be because one of the vectors is not getting transfected, even though i get a pretty decent transfection efficiency. Also, can only the GFP vector be transfected and not the others?

Suggestions and ideas would be immensely helpful

[bob1]

It is certainly posssible that there are different transfection efficiencies for each plasmid, though if you have optimised this transfection (i.e. amounts of each plasmid and total DNA) then this shouldn't be a problem.

The variations could be due to all sorts of things - you might be better off comparing % change between controls and treatments rather than trying to compare absolute values.