Hi,
I am new to promoter luciferase assay and i am facing a problem. I have tried both HepG2 and ZFL cell lines for this assay. My promoter plasmid (pGL3 basic+putative promoter) always produce lower signal in comparison to pGL3 basic. Could this be my promoter fragment hinders the basal expression of firefly in the pGL3 vector? But even it has the hindering effect, it should close to the pGL3 basic as pGL3 basic is promoterless, should not derive any expression of luciferase.
The assay is definitely needed to be further optimized but I do not know from where I should begin. Your opinion is highly appreciated. Thank you!
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