Nucleic Acid Hybrids, Formation and Structure of

Abstract

Hybridization is the formation of partially or completely double-stranded (duplex) nucleic acid (DNA:DNA, DNA:RNA, or RNA:RNA) by sequence-specific interaction of two complementary single-stranded nucleic acids. Reassociation or renaturation are often used to describe hybridization between completely complementary DNA or RNA strands. Solution hybridization is an integral part of the polymerase chain reaction. Hybridization is often carried out using a labeled probe in an attempt to detect the existence and quantity of complementary sequence in a complex nucleic acid mixture. The target is often immobilized, as in Southern and other blotting techniques. This chapter deals with the physical chemical aspects of hybridization, in particular, thermodynamics and kinetics. Melting temperatures, hybridization rates, and dissociation rates and temperatures are interrelated. Details are provided for calculation of these useful properties and examples of calculations are provided.