Microfluidic devices and methods of use in the formation and control of nanoreactors

摘要

The present invention provides novel microfluidic devices and methods that are useful for performing high-throughput screening assays and combinatorial chemistry. The invention provides for aqueous based emulsions containing uniquely labeled cells, enzymes, nucleic acids, etc., wherein the emulsions further comprise primers, labels, probes, and other reactants. An oil based carrier-fluid envelopes the emulsion library on a microfluidic device, such that a continuous channel provides for flow of the immiscible fluids, to accomplish pooling, coalescing, mixing, sorting, detection, etc., of the emulsion library.

主权项

1. A method for detecting a target nucleic acid fragment from a sample, the method comprising:
providing a sample comprising a plurality of different nucleic acid fragments; partitioning the sample into a plurality of droplets, wherein each of the plurality of droplets comprises one or more of the different nucleic acid fragments, reagents for an amplification reaction, and one or more different optically labeled probes specific for one or more different nucleic acid fragments; amplifying the different nucleic acid fragments in a plurality of droplets; flowing the plurality of droplets comprising the different amplified nucleic acid fragments past a detector that detects polarized light; detecting a level of polarized light emitted from each of the plurality of droplets comprising the different amplified nucleic acid fragments as each of the plurality of droplets flows past the detector; comparing the detected polarized light emitted from each of the plurality of droplets, comprising the different amplified nucleic acid fragments from the different portions of the sample in each of the plurality of droplets, to one another; and identifying the different amplified nucleic acid fragments from the different portions of the sample in one or more of the plurality of droplets in order to detect the different amplified nucleic acid fragments.