Detection of DNA hybridization using the TISPR-1 surface plasmon resonance biosensor.
Biotinylated oligonucleotide probes were immobilized to the gold sensor surface of the TISPR-1 miniature integrated surface plasmon resonance liquid sensor system for the purpose of detecting specific DNA hybridization. The immobilization of the oligonucleotide capture probes was carried out through streptavidin-biotin binding technology. The sensor detected the immobilization of unlabeled DNA through shifts in index of refraction as the molecules entered and remained selectively bound to the surface in the vicinity of the exponentially decaying surface plasmon resonance wave. The surface immobilization chemistry was proven to be stable for long periods of time, reproducible, and practical for detecting DNA hybridization with the TISPR-1. DNA hybridization was detected as a slow, positive, and small (when compared to protein-protein or antibody-antigen binding experiments) increase in the measured index of refraction under passive hybridization conditions by the TISPR-1 sensor. The DNA hybridization signal was significant (index of refraction change of 0.001) when large fragment PCR-amplified DNA products were hybridized to the oligonucleotide probes (S/N = 6-10). The DNA hybridization techniques were demonstrated using DNA sequences from the HIV genome which encode the Tat and Rev genes.
Kukanskis
K
K
Joint Program of Biomedical Engineering, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas, USA.
Elkind
J
J
Melendez
J
J
Murphy
T
T
Miller
G
G
Garner
H
H
D001483:Base Sequence; D015374:Biosensing Techniques; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D004867:Equipment Design; D016679:Genome, Viral; D006046:Gold; D006678:HIV / Q000235:genetics*; D008838:Microcomputers; D008904:Miniaturization; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides; D012984:Software; D020349:Surface Plasmon Resonance / Q000295:instrumentation* / Q000379:methods
D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D006046:Gold; D004247:DNA