Abstract: Compositions and methods for quantitative detection of target nucleic acids, such as miRNAs are disclosed. The methods are especially advantageous for single-color multiplex detection of two or more targets simultaneously (e.g., in the same reaction). The methods can involve optional reverse transcription followed by amplification performed with universal primers, fluorophore-labeled detection probes, and quencher oligonucleotides for quenching fluorescence of any detection probe not bound to a target molecule. The methods employ differential stability of detection probe-quencher oligonucleotide complexes, and by extension, differential fluorescence at various temperatures to distinguish between different target molecules.

Abstract: A method of sample analysis is provided. In some embodiments the method may comprise: hybridizing a tailed primer to a sample that comprises both wild type copies of a genomic locus and mutant copies of the genomic locus, extending the tailed primer using the genomic locus as a template to produce a primer extension product and detecting the primer extension product in a qPCR assay that employs a forward primer that is complementary to a sequence in the complement of the 5? tail of the tailed primer, a reverse primer, and a hydrolysis probe that is complementary to at least 6 nucleotides of the target complementary sequence of the tailed primer and at least 6 nucleotides of the 5? tail of the tailed primer. Kits for practice the method are also provided.

Abstract: Compositions and methods for quantitative detection of target nucleic acids, such as miRNAs are disclosed. The methods are especially advantageous for single-color multiplex detection of two or more targets simultaneously (e.g., in the same reaction). The methods can involve optional reverse transcription followed by amplification performed with universal primers, fluorophore-labeled detection probes, and quencher oligonucleotides for quenching fluorescence of any detection probe not bound to a target molecule. The methods employ differential stability of detection probe-quencher oligonucleotide complexes, and by extension, differential fluorescence at various temperatures to distinguish between different target molecules.

Abstract: Provided herein is a real-time PCR-based method of detecting, in a sample, an agent causing onychodystrophy, wherein the agent causing onychodystrophy belongs to a secondary clade member including one or more primary clade members. Also provided are compositions and kits that finds use in implementing the present method.

Abstract: Provided herein is a method of detecting an onychomycotic fungus in a sample, wherein the onychomycotic fungus belongs to a secondary clade member including one or more primary clade members. The method may include the steps of i) screening a sample using a first and second sets of secondary clade-specific primers to determine whether a secondary clade member among a plurality of secondary clade members is present or absent in the sample, where the plurality of secondary clade members includes (a) a dermatophyte, (b) a candida, and (c) a saprophyte, and ii) after determining the presence of the secondary clade member, screening the sample to determine whether an onychomycotic fungus is present or absent in the sample using primary clade-specific primers that are specific to a primary clade member that belongs to the secondary clade member. Also provided is a kit that finds use in implementing the present method.

Abstract: Provided herein is a real-time PCR-based method of detecting, in a sample, an agent causing onychodystrophy, wherein the agent causing onychodystrophy belongs to a secondary clade member including one or more primary clade members. Also provided are compositions and kits that finds use in implementing the present method.

Abstract: Provided herein is a method of detecting an onychomycotic fungus in a sample, wherein the onychomycotic fungus belongs to a secondary clade member including one or more primary clade members. The method may include the steps of i) screening a sample using a first and second sets of secondary clade-specific primers to determine whether a secondary clade member among a plurality of secondary clade members is present or absent in the sample, where the plurality of secondary clade members includes (a) a dermatophyte, (b) a candida, and (c) a saprophyte, and ii) after determining the presence of the secondary clade member, screening the sample to determine whether an onychomycotic fungus is present or absent in the sample using primary clade-specific primers that are specific to a primary clade member that belongs to the secondary clade member. Also provided is a kit that finds use in implementing the present method.

Abstract: Described herein are methods and assays relating to the detection of cMET alterations (e.g. variations in copy number and expression level, and/or the presence of mutations, including point mutations). Existing methods are limited in their clinical usefulness by, e.g., limited sensitivity, inter-lab discordance, or inability to provide the necessary multiplex ability. The methods and assays provided herein permit multimodal, multiplex assaying for faster, more cost-effective testing and screening of patients, permitting improved healthcare.

Abstract: Described herein are methods and assays relating to the detection of NRAS and/or BRAF alterations (e.g. variations in copy number and expression level, and/or the presence of mutations, including point mutations). Existing methods are limited in their clinical usefulness by, e.g., limited sensitivity, inter-lab discordance, or inability to provide the necessary multiplex ability. The methods and assays provided herein permit multimodal, multiplex assaying for faster, more cost-effective testing and screening of patients, permitting improved healthcare.

Abstract: The technology described herein relates to detecting the presence of one or more specific nucleic acids, e.g. determining the genotype of one or more allelic target site loci.