Effect of salt on the hybridization of DNA by sequential immobilization of oligonucleotides at the air-water interface in the presence of ODA/DOTAP monolayers

Abstract

The effect of low ionic strength on the binding of preformed DNA duplexes and the hybridization of single-stranded oligonucleotides at the air-water interface in the presence of cationic Langmuir monolayers of octadecylamine (ODA), as well as 1,2-dioleoyloxytrimethylammonium propane (DOTAP), is investigated. The complexation of the single-stranded DNA molecules and preformed duplexes with NaCl in solution with ODA/DOTAP Langmuir monolayers was followed in time by monitoring the pressure-area isotherms, wherein a very large and rapid expansion of the ODA/DOTAP monolayer was observed. In the case of sequential immobilization of complementary oligonucleotides, after addition of the complementary strand and intercalator, there was not much expansion, indicative of the fact that equilibrium had been rapidly achieved. Langmuir-Blodgett (LB) films of the ODA/DOTAP-DNA complex were formed on different substrates and characterized using quartz-crystal microgravimetry (QCM), fluorescence spectroscopy, and thermal melting studies. These measurements clearly showed that the preformed duplexes retained their native form as double helices and further, hybridization of the complementary single-stranded DNA molecules had occurred at the air-water interface, leading to the characteristic double-helical structure.