Kinetics of hybridization on the oligonucleotide microchips with gel pads.

Autor: Sorokin NV; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov str 32, 119991 Moscow, Russia., Chechetkin VR, Chechetkin MA, Vasiliskov VA, Turygin AY, Mirzabekov AD
Jazyk: angličtina
Zdroj: Journal of biomolecular structure & dynamics [J Biomol Struct Dyn] 2003 Oct; Vol. 21 (2), pp. 279-88.
DOI: 10.1080/07391102.2003.10506923
Abstrakt: The kinetics of hybridization on the oligonucleotide microchip with gel pads is studied both theoretically and experimentally. The monitoring of kinetics was performed with the measurements of fluorescence intensity produced by the labeled target oligonucleotides. As is shown, the hybridization time depends on the stability of the formed duplexes, the concentrations of target and probe oligonucleotides, and the diffusion of target oligonucleotides in solution and gel pad. The initial stage of hybridization is determined by the flow of target oligonucleotides from solution, then, followed by the diffusive propagation with approximately constant concentration of oligonucleotides at the boundary of gel pad and, finally, by the exponential saturation. The theoretical predictions of hybridization kinetics reveal a good correspondence with the experimental results and may be used for the choice of the optimal hybridization conditions. The possible applications of kinetic hybridization curves to the discrimination problems and assessment of diffusion coefficients in gel pads are briefly discussed. Finally, we discuss the relationships between the binding kinetics and the general functioning of biomolecular microchips.
Databáze: MEDLINE