Dual Luciferase Glow (Dual Glo)
November 27, 2012Download
Related Products: Gen5 Funktionen
Gen5 version required: 1.0 or higher
Basis for the Assay:
Genetic reporting assays are used to study gene expression and cellular responses to external stimuli. Dual-reporter assays use 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. This assay requires luminescence detection. “Glow” assays are typically less sensitive than “Flash” assays (see Dual-Luciferase® Reporter (DLR) Gene Assay), but don’t require automated injection and have a higher throughput.
The protocol executes the following sequence of steps:
- Read plate, 1 second/well (Read 1)
- Eject plate carrier and display message (add Renilla substrate)
- Read plate, 1 second/well (Read 2) A ratio (Read 1 / Read 2) is calculated for each well.
The experiment data file does not contain any data.
Plate configuration contains samples only, in duplicate. It may have to be updated with more specific information (e.g., standards to generate a calibration curve, blanks, controls).
The report provides:
- A matrix containing the raw data and Firefly/Renilla ratio
- Statistics on sample replicates