Abstract:
Method of analysis for alleles of a target. In the method, a plurality of fluid volumes may be formed. Each fluid volume may contain a first primer pair to amplify a first allele of a target, a second primer pair to amplify a second allele of the target, a first reporter including a first photoluminophore and providing a primer of the first primer pair, and a second reporter including a second photoluminophore and providing a primer of the second primer pair. Each of the first and second reporters may include an oligomer having a quencher, and the oligomer may be configured to base-pair with the primer of the first primer pair and the primer of the second primer pair. The first and second alleles may be amplified using the first and second primer pairs. Photoluminescence may be detected. A level of each allele may be determined.
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.
Abstract:
This disclosure provides methods for measuring the copy number for highly amplified and/or abundant genomic loci. Recognized herein is a need for methods for determining nucleic acid copy number, particularly in instances where one locus to be quantified (i.e., the target) is relatively more abundant than a locus of known abundance (i.e., the reference). In some cases, the method involves combining a query nucleic acid sample with a diluting nucleic acid sample and measuring the relative copy number of a target sequence compared with a reference sequence in the combined sample.
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.
Abstract:
System, including methods, apparatus, and compositions, for performing a digital assay with associated targets. In some embodiments, the assay of associated targets in partitions may lower the limit of detection (LOD) or otherwise increase assay sensitivity, accuracy, and/or specificity. The targets may represent spaced or overlapping regions of the same template and/or each may represent a region from a different complementary strand of the same template. In some embodiments, the associated targets may be provided by a type of biological particle, and the target content of partitions may be used to identify and quantify the type of biological particle in a sample.
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.
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.
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.
Abstract:
System, including methods, apparatus, and compositions, for performing a digital assay with associated targets. In some embodiments, the assay of associated targets in partitions may lower the limit of detection (LOD) or otherwise increase assay sensitivity, accuracy, and/or specificity. The targets may represent spaced or overlapping regions of the same template and/or each may represent a region from a different complementary strand of the same template. In some embodiments, the associated targets may be provided by a type of biological particle, and the target content of partitions may be used to identify and quantify the type of biological particle in a sample.