Highly Sensitive Northern Hybridization Using a Rapid Protocol for Downward Alkaline Blotting of RNA

Abstract

A simple and fast RNA gel blot procedureis described that uses 50 mM NaOH tosimultaneously transfer and fix RNA to apositively charged nylon membrane. TheRNA is transferred in a downward direction,and transfer is routinely completedwithin 2.5 h. The resulting blots give increasedsensitivity over existing methodswithout affecting RNA integrity and can beused in both radioactive and nonradioactivedetection procedures.