Kinetics of heterogeneous hybridization on indium tin oxide surfaces with and without an applied potential.
The rate of hybridization of oligonucleotide target sequences to chemically immobilized oligonucleotide probes has been studied both with and without an electrical field. The probe size was 20-24 nucleotides (nt) while the target size ranged from 157 to 864 nt. In agreement with previous studies, complete hybridization under normal conditions required 10-30 hours, depending on target size. The kinetics were characterized by a characteristic lag time followed by an asymptotic rise to the final value. In contrast, with an applied electrical field, all but the largest target hybridized in about 10 min while the longest hybridized within 1 h. Deleterious electrode reactions were avoided by close spacing of the anode and cathode and application of very small voltages. Our results suggest that probes and targets orient flat on the surface. A model is suggested to explain the kinetics observed that involves a series of surface states between initial target arrival and final hybridized state. Our results show that the electric field accelerated hybrid capture of solution-phase targets by surface-bound probes. This approach may have implications for enhancing array-based hybrid capture for mutation detection, copy number determination and/or gene expression profiling.
Department of Pediatrics, The Children's Hospital of Philadelphia, University of Pennsylvania, School of Medicine, Philadelphia, 19104, USA.
Hung-Ju
HJ
Su
Saul
S
Surrey
Steven E
SE
McKenzie
Paolo
P
Fortina
David J
DJ
Graves
D004247:DNA / Q000032:analysis*; D004058:Diffusion; D004553:Electric Conductivity; D007204:Indium; D007700:Kinetics; D009693:Nucleic Acid Hybridization / Q000379:methods*; D017971:Tin Compounds
D017971:Tin Compounds; D007204:Indium; D004247:DNA