Liquid-based hybridization assay with real-time detection in miniaturized array platforms.
An assay for the fluorescent detection of short oligonucleotide probe hybridization in miniaturized high-density array platforms is presented. It combines hybridization in solution with real-time fluorescent detection, which involves measurement of fluorescence increase by means of an induced fluorescence resonance energy transfer. The feasibility of this approach using DNA or RNA as a target, and short DNA- as well as LNA (locked nucleic acid)-modified oligonucleotides as probes is shown. The presented approach could potentially contribute to a significant increase in the throughput of large-scale genomic applications, such as oligofingerprinting and genotyping, and also reduce material consumption.
Guerasimova
Anna
A
Max Planck Institute for Molecular Genetics, Berlin, Germany.
Nyarsik
Lajos
L
Liu
Jian-Ping
JP
Schwartz
Regine
R
Lange
Matthias
M
Lehrach
Hans
H
Janitz
Michal
M
D003199:Computer Systems; D016172:DNA Fingerprinting / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation* / Q000379:methods; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation* / Q000379:methods; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008904:Miniaturization; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012996:Solutions
D012996:Solutions