Methods and systems for nucleic acid analysis
Inventors
SOSA, Jose Martin • Anthony, Joseph • Beckett, Nathan • Almogy, Gilad • Shee, Chandan • Lee, Phillip You Fai • WATANABE, Yuto
Assignees
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Publication Number
US-11904322-B2
Publication Date
2024-02-20
Expiration Date
Abstract
The present disclosure provides methods and processes for increasing the efficiency and accuracy of nucleic acid sequencing using techniques such as polymerase chain reaction (PCR). Methods and systems provided herein may facilitate performing reactions such as emulsion PCR (ePCR) on samples comprising nucleic acids and beads. The methods provided herein may provide a higher throughput compared to existing technologies.
Core Innovation
The disclosure relates to nucleic acid analysis systems and methods that perform polymerase chain reaction (PCR) using emulsion PCR. The system emphasizes high-throughput partition-based reactions, including droplets or partitions associated with beads, and downstream processing of the partitioned reaction mixture.
The disclosure further describes an emulsion breaking approach in which droplets are merged or coalesced in a container. A control system activates electrodes to alternate between positive and neutralizing ions at a predetermined frequency, and the temperature of the solution is changed or maintained at a predetermined temperature or temperature range sufficient to merge or coalesce the plurality of droplets.
The disclosure also describes a high-throughput nucleic acid workflow architecture that incorporates modular stages. The stages include emulsion formation, thermocycling, emulsion breaking, washing/enrichment, and sequencing, with processing performed in cartridge or well array configurations and with parallel or continuous batch processing controlled by a computer system.
Claims Coverage
The provided independent-claim content includes one droplet-processing system claim. It covers a container with droplets in contact with surfaces, electrodes in electrical communication with the surfaces, and a heat exchange system, with a controller that activates alternating positive and neutralizing ions at a predetermined frequency and changes or maintains temperature to merge or coalesce droplets.
Droplet merging or coalescence using alternating positive and neutralizing ions with heat exchange control
A system with a container containing a plurality of droplets in contact with first and second surfaces, electrodes in electrical communication with the first and second surfaces, and a heat exchange system thermally coupled to a third surface, wherein a controller simultaneously activates the first and second electrodes to alternate between positive and neutralizing ions at a predetermined frequency and changes or maintains a solution temperature at a predetermined temperature or temperature range, with the predetermined frequency and/or predetermined temperature or temperature range sufficient to merge or coalesce the plurality of droplets.
The main inventive coverage is a droplet-processing system that combines alternating positive/neutralizing-ion electrode activation at a predetermined frequency with heat exchange temperature control to merge or coalesce droplets in a container.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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