Chromophore-based PCR methods for detecting and characterizing nucleic acids in samples
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United States of America Patent
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Sep 25, 2018
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Feb 1, 2018
app pub date -
Aug 11, 2017
filing date -
Jan 24, 2013
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Abstract
Methods of detecting at least one genetic variation in a polynucleotide analyte in a sample. A fluorophore is attached to a first primer, a quencher is attached to a second primer, the first primer and the second primer are specific for the polynucleotide analyte. A signal generated by the fluorophore and quencher is measured. PCR is performed with the first primer and the second primer using the polynucleotide analyte as a template, thereby amplifying the template. A signal generated by the fluorophore and quencher from the PCR amplification product is measured. Comparison is made of the signals; and a determination is made of the presence or absence of the at least one genetic variation based i) on the change in signal as determined; and ii) by comparing said change to the change in signal observed upon PCR amplification for a corresponding polynucleotide analyte lacking the at least one genetic variation.
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