A method of identifying a first target nucleic acid comprising, providing a sample comprising the first target nucleic acid, providing a first set of paired oligonucleotides with complementarity to the first target nucleic acid, the first set of paired oligonucleotides comprising a first ratio of (a) first competitive oligonucleotides to (b) first signal oligonucleotides comprising a signal tag, wherein the competitive oligonucleotides compete with the signal oligonucleotides for binding to the first target nucleic acid, amplifying the first target nucleic acid with the polymerase chain reaction, thereby degrading the first signal oligonucleotide and permitting generation of a first signal, generating the first signal, measuring intensity of the first signal, and correlating the intensity of the first signal to the first ratio, thereby identifying the first target nucleic acid.

Provided are methods of amplifying nucleic acids. The methods include combining a nucleic acid sample, a known amount of one or more competitive internal standard nucleic acids, and one or more amplification primers adapted to amplify one or more nucleic acids of interest present in the nucleic acid sample and the one or more competitive internal standard nucleic acids. The nucleic acid sample, competitive internal standard nucleic acids, and amplification primers are combined in a reaction mixture under conditions sufficient to amplify the one or more nucleic acids of interest and the one or more competitive internal standard nucleic acids. Aspects of the present disclosure further include compositions and kits that find use in practicing embodiments of the methods.

Provided are methods of amplifying nucleic acids. The methods include combining a nucleic acid sample, a known amount of one or more competitive internal standard nucleic acids, and one or more amplification primers adapted to amplify one or more nucleic acids of interest present in the nucleic acid sample and the one or more competitive internal standard nucleic acids. The nucleic acid sample, competitive internal standard nucleic acids, and amplification primers are combined in a reaction mixture under conditions sufficient to amplify the one or more nucleic acids of interest and the one or more competitive internal standard nucleic acids. Aspects of the present disclosure further include compositions and kits that find use in practicing embodiments of the methods.

Methods and compositions for detecting an allelic form of a target. In an exemplary method, partitions may be created that collectively contain at least one first allelic form and a second allelic form of a target. Each partition may contain (i) a same probe capable of binding specifically to each of the first and second allelic forms of the target and (ii) a competitor configured to bind selectively to the second allelic form relative to the first allelic form and to block binding of the probe to the second allelic form. The first allelic form of the target may be amplified in the partitions. A signal may be detected from a label of the probe while the label is contained by the partitions. A number of partitions that are positive (or negative) for the at least one first allelic form may be determined based on the signal.

Methods and compositions for detecting an allelic form of a target. In an exemplary method, partitions may be created that collectively contain at least one first allelic form and a second allelic form of a target. Each partition may contain (i) a same probe capable of binding specifically to each of the first and second allelic forms of the target and (ii) a competitor configured to bind selectively to the second allelic form relative to the first allelic form and to block binding of the probe to the second allelic form. The first allelic form of the target may be amplified in the partitions. A signal may be detected from a label of the probe while the label is contained by the partitions. A number of partitions that are positive (or negative) for the at least one first allelic form may be determined based on the signal.

Embodiments of a method and/or system can include generating a set of target-associated molecules (e.g., spike-in molecules) associated with one or more biological targets; generating one or more spike-in mixtures based on processing the set of target-associated molecules with one or more samples including the one or more biological targets; performing one or more Sanger sequencing operations on the one or more spike-in mixtures; determining one or more abundance metrics based on chromatogram-related outputs from the one or more Sanger sequencing operations; and/or facilitating characterization of one or more medical conditions based on the one or more abundance metrics.

Embodiments of a method and/or system can include generating a set of target-associated molecules (e.g., spike-in molecules) associated with one or more biological targets; generating one or more spike-in mixtures based on processing the set of target-associated molecules with one or more samples including the one or more biological targets; performing one or more Sanger sequencing operations on the one or more spike-in mixtures; determining one or more abundance metrics based on chromatogram-related outputs from the one or more Sanger sequencing operations; and/or facilitating characterization of one or more medical conditions based on the one or more abundance metrics.

Described herein are compositions for evaluating nucleic acids, standardized mixture for assessing amounts of at least one target nucleic acid in a sample, and kits comprising the same.

Described herein are compositions for evaluating nucleic acids, standardized mixture for assessing amounts of at least one target nucleic acid in a sample, and kits comprising the same.

The invention features compositions and methods that are useful for the measurement of the quantity of a nucleic acid target in a sample.