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AssigneeKasa Bio, L.L.C.
InventorsKiran Madanahally Divakar, Shashi Bala
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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.
A method for detection of two or more distinct target polynucleotides in a sample the method comprising performing one or more amplification cycles on the sample using a polymerase and universal or target-specific forward and reverse primers to generate amplified products corresponding to the target polynucleotides, and detecting and/or quantifying each amplified product using a plurality of distinct unbound detection probes and a plurality of corresponding quencher oligonucleotides that can hybridize thereto.
A method for detection of target miRNA, the method comprising:
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