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US 9,266,104 B2Granted
Thermocycling device for nucleic acid amplification and methods of use
AssigneeRaindance Technologies, Inc.
InventorDarren Roy Link
Life of this patent
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Abstract
The present invention provides thermocycling devices useful for amplification of nucleic acids in droplets. The thermocycling device utilizes the flow of one or more fluids through a main compartment at temperatures sufficient to conduct a polymerase chain reaction. Methods of amplifying nucleic acids in droplets are also provided.
Claims21
A thermocycling device for amplifying nucleic acid in a droplet, the device comprising:
- a reaction chamber comprising a plurality of droplets;
- a first temperature-controlled fluid source comprising a fluid at a first temperature;
- a second temperature-controlled fluid source comprising a fluid at a second temperature;
- and
- a conduit fluidly coupled to the first and the second temperature-controlled fluid sources, and the reaction chamber;
- wherein the device is adapted to immerse the plurality of droplets in the fluid at the first temperature within the reaction chamber and subsequently immerse the plurality of droplets in the fluid at the second temperature within the reaction chamber, wherein the reaction chamber is oriented so that a gravitational force causes the fluid at the first temperature and the fluid at the second temperature to exit the reaction chamber while maintaining the plurality of droplets in the reaction chamber.
3 more elements
A method of nucleic acid amplification, said method comprising the steps of:
- a) providing a reaction chamber for housing a plurality of droplets;
- b) flowing the plurality of droplets into the reaction chamber, each droplet comprising a single nucleic acid template, at least one primer and reagents sufficient for nucleic acid amplification;
- c) flowing a first fluid having a first temperature into the reaction chamber and maintaining for a sufficient time to denature the nucleic acid template in the droplets, wherein the reaction chamber is oriented so that a gravitational force causes the first fluid to exit the reaction chamber while maintaining the droplets in the reaction chamber;
- d) flowing a second fluid having a second temperature into the reaction chamber and maintaining for a sufficient time to anneal one or more of the PCR reagents to the nucleic acid template in the droplets, wherein the reaction chamber is oriented so that a gravitational force causes the second fluid to exit the reaction chamber while maintaining the droplets in the reaction chamber;
- e) flowing a third fluid having a third temperature into the reaction chamber and maintaining for a sufficient time to extend the nucleic acid template in the droplets, wherein the reaction chamber is oriented so that a gravitational force causes the third fluid to exit the reaction chamber while maintaining the droplets in the reaction chamber.
2 more elements
Dependencies are inferred from the claim text.