Dot-Blot Hybridization Method

Nucleic acid hybridization is a very potent technique that can be used for the identification of DNA and RNA species with varying degree of homology and for the estimation of relative amounts of nucleic acid with known homolgy. In most cases, single-stranded (ss) (denatured) DNA or RNA is bound to a filter support (e.g., nitrocellulose) and incubated with a radioactively labeled ss DNA or RNA fragment (“probe”) complementary to the nucleic acids of interest. In subsequent washing steps the nonhybridized probe is removed. Hybridized probe will be retained, but may be washed off in further washing steps, depending on the ion concentration and temperature of the wash.