Statistical Thermodynamics and Kinetics of DNA Multiplex Hybridization Reactions

¹ Portland State University

^* To whom correspondence should be addressed. E-mail: abenight{at}pdx.edu.

Submitted on June 8, 2006
Revised on July 17, 2006
Accepted on 26 July 2006

	Abstract

A general analytical description of the equilibrium and reaction kinetics of DNA multiplex hybridization has been developed. In this approach, multiplex hybridization is considered to be a competitive, multi-channel, reaction process: a system wherein many species can react both specifically and non-specifically with one another. General equations are presented that can consider equilibrium and kinetic models of multiplex hybridization systems comprised, in principle, of any number of targets and probes. Numerical solutions to these systems for both equilibrium and kinetic behaviors are provided. Practical examples demonstrate clear differences between results obtained from more common simplex methods, in which individual hybridization reactions are considered to occur in isolation; and multiplex hybridization, where desired and competitive cross-hybrid reactions between all possible pairs of strands are considered. In addition sensitivities of the hybridization process of the perfect match duplex, to temperature, target concentration, and existence of sequence homology with other strands, are examined. This general approach also considers explicit sequence dependent interactions between targets and probes involved in the reactions. Sequence dependent stabilities of all perfect match and mismatch duplex complexes are explicitly considered and effects of relative stability of cross-hybrid complexes are also explored. Results reveal several interdependent factors that strongly influence DNA multiplex hybridization behavior. These include: relative concentrations of all probes and targets; relative thermodynamic stability of all perfect match and mismatch complexes; sensitivity to temperature, particularly for mismatches and; amount of sequence homology shared by the probe and target strands in the multiplex mix.

Key Words: DNA, Hybridization, Kinetics, MIcroarrays, Multiplexing, Thermodynamics

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