Nucleic acid hybridization reactions are at the heart of a number of powerful assays used in molecular biology research and increasingly in diagnostic applications. Hybridization reactions typically require long incubation times, i.e. overnight. Recent work has demonstrated that a ribonucleoprotein and certain cationic detergents can increase hybridization rates several thousand fold. The overall goal of this project is to use this technology to develop high-speed nucleic acid hybridization procedures for use in solution and solid phase assays such as Southern/Northern blotting. During Phase l we were successful in accelerating RNA:RNA solution hybridization reactions in ribonuclease protection assay format such that reactions which normally require 24 hours, were complete in less than 30 seconds. We also demonstrated the feasibility of using cationic detergents to accelerate hybridization reactions of nucleic acids immobilized on beads and on dot blots. During Phase Il we will refine conditions for solution hybridizations and develop protocols for membrane based hybridization procedures such as Southerns and Northerns. The successful completion of this project will have major impact, both on the research and the diagnostic applications of nucleic acid based hybridization assays.
