DIGITAL ANALYTE ANALYSIS
First Claim
Patent Images
1. A method for analyzing a target nucleic acid, the method comprising the steps of:
- forming droplets containing a single target nucleic acid and one or more amplification reagents;
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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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.
210 Citations
23 Claims
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1. A method for analyzing a target nucleic acid, the method comprising the steps of:
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forming droplets containing a single target nucleic acid and one or more amplification reagents; 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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method for processing nucleic acid samples, the method comprising the steps of:
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obtaining droplets comprising one nucleic acid target; amplifying said nucleic acid in said droplets; and separating droplets that comprise a heterogeneous population of amplicons from droplets containing a homogeneous population of amplicons.
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16. A method for detecting a recurrence of a cancer in a patient, the method comprising:
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forming sample droplets wherein, on average, each droplet comprises a single target nucleic acid derived from a patient sample; flowing the sample droplets through a channel; amplifying the target in the droplets; detecting amplified target in the droplets; excluding droplets comprising a heterogeneous population of amplicons; and analyzing non-excluded droplets to determine the presence of mutant alleles indicative of recurrence. - View Dependent Claims (17, 18, 19, 20)
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21. A method for analyzing a target nucleic acid, the method comprising the steps of:
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compartmentalizing a first fluid into portions, each portion containing a single target nucleic acid; amplifying the target in the portions; excluding portions containing amplicon from the target and amplicon from a variant of the target; and analyzing target amplicons. - View Dependent Claims (22)
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23. A method for analyzing a target nucleic acid, the method comprising the steps of:
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forming sample droplets containing 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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Specification