Assuntos
Técnicas Genéticas , Genótipo , Polimorfismo de Nucleotídeo Único , Alelos , Interpretação Estatística de Dados , Técnicas Genéticas/instrumentação , Técnicas Genéticas/normas , Técnicas Genéticas/estatística & dados numéricos , Variação Genética , Humanos , Reação em Cadeia da Polimerase , Controle de Qualidade , RNA Mensageiro/genéticaRESUMO
A high-density fiber-optic DNA microarray sensor was developed to monitor multiple DNA sequences in parallel. Microarrays were prepared by randomly distributing DNA probe-functionalized 3.1-microm-diameter microspheres in an array of wells etched in a 500-microm-diameter optical imaging fiber. Registration of the microspheres was performed using an optical encoding scheme and a custom-built imaging system. Hybridization was visualized using fluorescent-labeled DNA targets with a detection limit of 10 fM. Hybridization times of seconds are required for nanomolar target concentrations, and analysis is performed in minutes.
Assuntos
DNA/química , Sequência de Bases , Tecnologia de Fibra Óptica , Microesferas , Hibridização de Ácido Nucleico , Sondas de Oligonucleotídeos , Fibras Ópticas , Sensibilidade e EspecificidadeRESUMO
We have developed a randomly ordered fiber-optic gene array for rapid, parallel detection of unlabeled DNA targets with surface immobilized molecular beacons (MB) that undergo a conformational change accompanied by a fluorescence change in the presence of a complementary DNA target. Microarrays are prepared by randomly distributing MB-functionalized 3-microm diameter microspheres in an array of wells etched in a 500-microm diameter optical imaging fiber. Using several MBs, each designed to recognize a different target, we demonstrate the selective detection of genomic cystic fibrosis related targets. Positional registration and fluorescence response monitoring of the microspheres was performed using an optical encoding scheme and an imaging fluorescence microscope system.
