Methods and systems for nucleic acid analysis and quantification
AssigneeCombinati Incorporated
InventorsJu-Sung Hung, Megan Dueck, Andrew Zayac
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Abstract
The present disclose provides methods and systems for amplifying and quantifying nucleic acids and for detecting the presence or absence of a target in a sample. The methods and systems provided herein may utilize a device comprising a plurality of partitions separated from an external environment by a gas-permeable barrier. Certain methods disclosed herein involve subjecting nucleic acid molecules in the plurality of partitions to conditions sufficient to conduct nucleic acid amplification reactions. The nucleic acid molecules may be subjected to controlled heating in the plurality of partitions to generate data indicative of a melting point(s) of the nucleic acid molecules.
Claims21
A method for analyzing a plurality of nucleic acid molecules, comprising:
- (a) providing a device comprising a plurality of partitions, wherein at least a subset of said plurality of partitions comprises said plurality of nucleic acid molecules, wherein each partition of said at least said subset of said plurality of partitions is configured to permit gas flow from said at least said subset of said plurality of partitions to an environment external to said at least said subset of said plurality of partitions through at least one thin film separating said at least said subset of said plurality of partitions from said external environment, wherein said at least one thin film is (i) substantially gas impermeable when a first pressure differential is applied across said at least one thin film and (ii) substantially gas permeable when a second pressure differential greater than said first pressure differential is applied across said at least one thin film;
- (b) using a fluid flow unit to apply a pressure differential across said at least one thin film to subject gas in said plurality of partitions to flow through said at least one thin film to said external environment;
- (c) while subjecting said at least said subset of said plurality of partitions to controlled heating, collecting signals from said at least said subset of said plurality of partitions;
- and
- (d) processing said signals collected in (c) to yield data indicative of at least one melting point of at least a subset of said plurality of nucleic acid molecules in said at least said subset of said plurality of partitions.
A method for analyzing a plurality of nucleic acid molecules, comprising:
- (a) providing a device comprising a plurality of partitions, wherein at least a subset of said plurality of partitions comprises said plurality of nucleic acid molecules, wherein each partition of said at least said subset of said plurality of partitions is configured to permit gas flow from said at least said subset of said plurality of partitions to an environment external to said at least said subset of said plurality of partitions through at least one thin film separating said at least said subset of said plurality of partitions from said external environment, wherein said at least one thin film is (i) substantially gas impermeable when a first pressure differential is applied across said at least one thin film and (ii) substantially gas permeable when a second pressure differential greater than said first pressure differential is applied across said at least one thin film;
- (b) using a fluid flow unit to apply a pressure differential across said at least one thin film to subject gas in said plurality of partitions to flow through said at least one thin film to said external environment;
- (c) subjecting said at least said subset of said plurality of partitions to conditions sufficient to conduct nucleic acid amplification reactions using said plurality of nucleic acid molecules to generate amplification products from at least a subset of said plurality of nucleic acid molecules;
- (d) while subjecting said at least said subset of said plurality of partitions to said conditions in (c), collecting signals from said at least said subset of said plurality of partitions over a plurality of time points;
- and
- (e) processing said signals to determine a number of nucleic acid molecules in said at least said subset of said plurality of partitions.
Dependencies are inferred from the claim text.