Ruediger, R., Brewis, N., Ohst, K. and Walter, G.
Notes: This paper describes a method for quantitation of luciferase mRNA by in vitro translation using Promega TNT® Coupled Wheat Germ Extract System. (Rabbit reticulocyte lysate could not be used because of luciferase quenching problems). In this reporter gene assay, COS cells were transfected with a firefly luciferase reporter plasmid driven by promoters/enhancers of varying strengths and total cellular RNA was isolated and translated in vitro using the TNT® System. To normalize expression, a CMV Renilla luciferase or beta-galactosidase vector was used as a control. To measure luciferase activity either Promega Luciferase Assay System or Dual-Luciferase® Reporter Assay System was used. (2047)