Nucleic Acid Hybridization. A Theoretical Consideration

Abstract

DNA-DNA and RNA-DNA hybridization reactions are the basis of many assays in DNA analysis and are presently some of the most frequently used techniques in molecular biology. The hybridization reaction is the formation of partial or complete double-stranded nucleic acid molecules by sequence-specific interaction of two complementary single-stranded nucleic acids. The hybridization reaction, using labeled probes, is the only practical way to detect the presence of specific nucleic acid sequences in a complex nucleic acid mixture. The most frequently used hybridization technique is the membrane hybridization technique. Denatured DNA or RNA is immobilized on an inert support in a way that self-annealing is prevented but bound sequences are available for hybridization with labeled single or double stranded probes. Extensive washing of the membrane to remove unbound probe and poorly matched hybrids follows the hybridization reaction. Membrane hybridization is used in many different applications such as Southern and Northern blot hybridization, dot blot hybridization and phage plaque or bacterial colony hybridization. This chapter will briefly consider theoretical aspects of membrane hybridization.