AMPLIFICATION REPORTER WITH BASE-PAIRING OLIGOMERS
First Claim
1. A method of analysis, the method comprising:
- forming at least one volume containing a reporter including a first oligomer and a second oligomer capable of base-pairing with one another below a melting temperature of the reporter to affect a photoluminescence detectable from the reporter;
amplifying a target in the at least one volume, at least in part by extending one or more primers at a temperature above the melting temperature of the reporter;
detecting the photoluminescence of the reporter from the at least one volume while the at least one volume is at a temperature below the melting temperature of the reporter; and
determining a property of the target based on the photoluminescence detected.
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Accused Products
Abstract
System, including methods, apparatus, and compositions, for performing amplification assays with an amplification reporter including a first oligomer and a second oligomer capable of base-pairing with one another below a melting temperature of the reporter. The reporter may have a detectable photoluminescence that is affected, such as reduced, by base-pairing of the first and second oligomers with one another. A target, such as a nucleic acid target sequence, may be amplified in at least one volume, such as a plurality of partitions, above the melting temperature, and photoluminescence of the reporter may be detected from the at least one volume below the melting temperature. A property of the target, such as a concentration of the target, may be determined based on the photoluminescence detected.
13 Citations
20 Claims
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1. A method of analysis, the method comprising:
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forming at least one volume containing a reporter including a first oligomer and a second oligomer capable of base-pairing with one another below a melting temperature of the reporter to affect a photoluminescence detectable from the reporter; amplifying a target in the at least one volume, at least in part by extending one or more primers at a temperature above the melting temperature of the reporter; detecting the photoluminescence of the reporter from the at least one volume while the at least one volume is at a temperature below the melting temperature of the reporter; and determining a property of the target based on the photoluminescence detected. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A method of analysis, the method comprising:
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forming partitions each containing a reporter including a first oligomer having a photoluminophore and also including a second oligomer capable of base-pairing with the first oligomer below a melting temperature of the reporter to decrease, by energy transfer, a photoluminescence detectable from the photoluminophore; exposing the partitions to a plurality of thermal cycles, to amplify a target sequence at least in part by extending one or more primers at a temperature above the melting temperature of the reporter, wherein the target sequence is present in only a subset of the partitions; detecting the photoluminescence of the photoluminophore for each partition of a plurality of the partitions while the partition is at a temperature below the melting temperature of the reporter; and determining a property of the target based on the photoluminescence detected. - View Dependent Claims (15, 16, 17, 18, 19)
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20. A composition, comprising:
a plurality of partitions containing a target at partial occupancy and amplification reagents to amplify the target and also containing a reporter to detect target amplification, the reporter including a first oligomer having a photoluminophore and a second oligomer that is base-paired with the first oligomer to affect a photoluminescence detectable from the photoluminophore.
Specification