Issue October 2003

category image Volume 21
No. 2 (p 159-310)
October 2003
ISSN 0739-1102

Kinetics of Hybridization on the Oligonucleotide Microchips with Gel Pads (p. 279-288)

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.

N. V. Sorokin1,2
V. R. Chechetkin1,3,*
M. A. Livshits1
V. A. Vasiliskov1
A. Y. Turygin1
A. D. Mirzabekov1

1Engelhardt Institute of Molecular Biology
Russian Academy of Sciences
Vavilov str. 32
119991 Moscow, Russia
2Moscow Institute of Physics & Technology
141700 Dolgoprudny, Russia
3Troitsk Institute of Innovation and Thermonuclear Investigations (TRINITI)
Theoretical Department of Division for Perspective Investigations
142190 Troitsk
Moscow Region, Russia
*chechet@eimb.ru
vladimir_chechet@mail.ru

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