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US 2012/0021423 A1Published application
Controls and calibrators for tests of nucleic acid amplification performed in droplets
AssigneeQuantalife, Inc.
InventorsBilly Wayne Colston, JR., Benjamin Joseph Hindson, Kevin Dean Ness, Donald Arthur Masquelier
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Abstract
System, including methods and apparatus, for performing droplet-based tests of nucleic acid amplification that are controlled and/or calibrated using signals detected from droplets.
Claims48
A method of performing a droplet-based assay, comprising:
- detecting a signal from each of a plurality of droplets;
- comparing a width of the signal from each droplet to a permitted range;
- excluding droplets for which the signal has a width that is not in the permitted range, to identify a set of included droplets;
- and
- determining a concentration of a target provided by a sample disposed in the plurality of droplets using data collected from the included droplets and without any contribution of data collected from the excluded droplets.
2 more elements
A method of performing a droplet-based assay, comprising:
- detecting a signal from at least two types of calibration droplets, the signal for each type of calibration droplet being of different intensity;
- detecting sample data from sample droplets;
- and
- determining if amplification of a target occurred in each of the sample droplets based on the sample data and the signal of each different intensity detected from the calibration droplets.
1 more element
A method of performing a droplet-based assay, comprising:
- generating droplets from an aqueous phase including a first dye and a second dye, the second dye being an internal reference;
- detecting sample data from the first dye included in the droplets, the sample data being related to a reaction performed in the droplets;
- detecting reference data from the second dye included in the droplets;
- transforming the sample data with the reference data to reduce variability in the sample data that is independent of the reaction;
- and
- determining if the reaction occurred in each of the sample droplets based on sample data that has been transformed with the reference data.
3 more elements
A method of performing a droplet-based assay, comprising:
- detecting a signal from each of a plurality of droplets flowing through a detection region;
- transforming an intensity of the signal for each of the plurality of droplets according to a duration of such signal to obtain transformed signals;
- and
- determining whether amplification of a target occurred in individual droplets based on the transformed signals.
1 more element
A method of performing a droplet-based assay, comprising:
- generating droplets from an aqueous phase including a sample and first and second dyes;
- detecting sample data from the first dye in the droplets, the sample data being related to amplification of a test target from the sample;
- detecting control data from the second dye in the droplets, the control data being related to amplification of a control target in individual droplets;
- analyzing the sample data and the control data to determine respective concentrations of the test target and the control target;
- and
- correlating the concentration of the test target with the concentration of the control target.
3 more elements
A method of performing a droplet-based assay, comprising:
- obtaining a first set of droplets configured to amplify a test target from a sample disposed in the first set of droplets, and a second set of droplets configured to amplify a control target in the second set;
- detecting test amplification data from the first set of droplets and control amplification data from the second set of droplets;
- and
- analyzing the test amplification data and the control amplification data to determine a concentration of the test target and the control target;
- and
- correlating the concentration of the test target with the concentration of the control target.
3 more elements
Dependencies are inferred from the claim text.