Digital analyte analysis
First Claim
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1. A method for detecting target sequences from a sample, the method comprising:
- providing a plurality of droplets each comprising a nucleic acid molecule, reagents for an amplification reaction, and a plurality of different optically labeled probes, each specific for a different target sequence;
amplifying a target sequence from the nucleic acid molecule in the droplets;
flowing the droplets past a detector that detects polarized light; and
detecting a level of polarized light emitted from an optically labeled probe that identifies the target sequence in each of the droplets.
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Abstract
The invention generally relates to droplet based digital PCR and methods for analyzing a target nucleic acid using the same. In certain embodiments, methods of the invention involve forming sample droplets containing, on average, a single target nucleic acid, amplifying the target in the droplets, excluding droplets containing amplicon from the target and amplicon from a variant of the target, and analyzing target amplicons.
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24 Claims
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1. A method for detecting target sequences from a sample, the method comprising:
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providing a plurality of droplets each comprising a nucleic acid molecule, reagents for an amplification reaction, and a plurality of different optically labeled probes, each specific for a different target sequence; amplifying a target sequence from the nucleic acid molecule in the droplets; flowing the droplets past a detector that detects polarized light; and detecting a level of polarized light emitted from an optically labeled probe that identifies the target sequence in each of the droplets. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 19, 20, 21, 22, 23, 24)
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9. A method for detecting target sequences from a sample, the method comprising:
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providing a plurality of droplets each comprising a nucleic acid molecule, reagents for an amplification reaction, and a plurality of different detectably labeled probes, each specific for a different target sequence; amplifying a target sequence from the nucleic acid molecule in the droplets; configuring a polarization filter with respect to an excitation polarization; flowing the droplets past a detector that comprises the polarization filter; and detecting a level of polarized light emitted from a detectably labeled probe that identifies the target sequence in each of the droplets. - View Dependent Claims (10, 11, 12, 13, 14, 15, 16, 17, 18)
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Specification