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Luciferase assay - extreme variation in RLU

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#1 Pkrish



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Posted 01 March 2013 - 08:40 AM


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

#2 bob1


    Thelymitra pulchella

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Posted 02 March 2013 - 12:01 AM

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.

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