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Genetic reporting assays are widely used to study gene expression and cellular responses to external stimuli in prokaryotic and eukaryotic organisms. Dual-reporter assays, as the name implies entails the use of two independent reporter systems simultaneously to improve experimental accuracy. One reporter is usually tied to measuring the response resulting from the experimental conditions and is often referred to as the “experimental” reporter. The other reporter is designed such that it does not respond to the experimental conditions, acting as an internal control from which data generated by the experimental reporter can be normalized to. Normalization of the data serves to compensate for variability caused by differences in transfection efficiency, cell viability, cell lysis, and pipetting. Promega’s Dual-Luciferase® system uses the activities of luminescent proteins (luciferases) from the firefly (Photinus pyralis) beetle and the sea pansy (Renilla renformis) to serve as an experimental and a control reporter, respectively. Here we describe the use of the Clarity™ Luminescence Microplate Reader (BioTek Instruments) to perform dual-luciferase measurements with purified recombinant enzymes.
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