Evaluation of hybridization kinetics of molecular beacons with nucleic acids using picoliter microfluidic droplet with millisecond resolution

A microfluidic droplet analysis platform is developed for evaluating the hybridization kinetics of nucleic acid assays. In this platform molecular beacons (MB) and different sequence of nucleic acids are rapidly mixed in picoliter droplets and flown one-by-one through microchannels. Specific nucleic acid hybridization is evaluated by the accumulated fluorescence emitted from the droplets. This process does not require immobilization and stringent washing processes and is highly sensitive compared to conventional nucleic acid detection methods. The hybridization rates of MB with nucleic acids are determined by the mixing efficiency in picoliter-sized microdroplets. The hybridization kinetics is observed at millisecond resolutions. This nucleic acid assay platform can be the basis for an integrated rapid nucleic acid detection system that is programmable and requires only a few micro liters of sample