Multiplexing to Improve Optimization
Tracking cell viability along with reporter activity is critical to determining optimal transfection conditions. High reporter activity may come at the expense of cell health, and unhealthy cells are less likely to exhibit consistent, physiologically relevant responses.
Optimization is simplified further by using reporter and viability assays that can be measured in the same sample. The ONE-Glo™ Luciferase Assay System and the CellTiter-Fluor™ Cell Viability Assay are compatible assays. By multiplexing these two assays, both reporter activity and viability can be measured in the same well of a 96-well plate in less than an hour. No medium changes or washing are needed.
Empirically determining optimal transfection conditions for any given cell type allows for the best return from your cell-based experiments. Optimal conditions should yield maximum reporter activity with minimum impact on cell health, thus preserving the biology of the cells for subsequent manipulation. By understanding the keys to successfully transfecting plasmid DNA, following a standard optimization plate layout, and multiplexing reporter and viability assays, optimal parameters can be determined with relative ease.