sepharose and methylmercury-hydroxide

We describe a method for hybridization of cDNA probes to RNA directly in agarose gels which provides a practical alternative to methods involving transfer of the RNA out of the gel. Total cellular RNA is subjected to electrophoresis in agarose gels containing methylmercuric hydroxide as the denaturing agent. After removal of the methylmercuric hydroxide, the gel is dried and 32P-labeled DNA probes are hybridized to the immobilized RNA. This method is more economical in time and expense than methods involving transfer of the RNA out of the gel, while maintaining a level of sensitivity comparable to other procedures.

Topics: Animals; Chick Embryo; DNA; Electrophoresis, Agar Gel; Methods; Methylmercury Compounds; Nucleic Acid Hybridization; RNA; Sepharose