DETECTION AND QUANTITATION OF SPECIFIC MESSENGER-RNAS BY RIBONUCLEASEPROTECTION ASSAY USING DENATURING HORIZONTAL POLYACRYLAMIDE-GEL ELECTROPHORESIS - A RADIOACTIVE AND NONRADIOACTIVE APPROACH

A radioactive (P-32) and nonradioactive (digoxigenin) ribonuclease pro tection assay (RPA) has been developed to detect mRNAs of housekeeping proteins and growth factors. A modification of polyacrylamide gel ele ctrophoresis (PAGE) to simplify RPA is described. Both Cleangels (Phar macia) and laboratory-cast polyacrylamide gels, in a denaturing, horiz ontal electrophoresis system, were used. The amount of toxic chemicals and waste was reduced, in comparison with sequencing gels normally us ed for RPA. The protected RNA fragments were shown to be well-separate d, with sufficient sensitivity in this modified, quick gel system.