KINETICS OF DNA HYBRIDIZATION WITH OLIGONUCLEOTIDES IMMOBILIZED IN GEL LAYER

Theoretical analysis of the kinetics of DNA hybridization with oligonu cleotides immobilized in apolyacrylamide gel layer showed that establi shment of the equilibrium uniform DNA distribution is governed by the mechanism of retarded diffusion; the equilibration time is longer for more stable duplexes than for less stable ones. Conditions for optimal discrimination of perfect duplexes from those with mismatches are ana lyzed. The hybridization process should be brought to equilibrium unde r low-temperature nonsaturating conditions. The kinetic differences be tween perfect and imperfect duplexes allow further contrasting of the hybridization pattern through additional washing at low temperature of appropriate duration after hybridization.