METHOD, PRODUCT, AND SYSTEM FOR QUANTIFYING THE METHYLATION STATUS OF A NUCLEIC ACID IN A SAMPLE
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United States of America Patent
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Sep 18, 2014
app pub date -
Mar 14, 2014
filing date -
Mar 15, 2013
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Abstract
Methods and systems are described for quantifying the methylation status of nucleic acids in a sample utilizing standardized curves derived from methylation-sensitive HRM data. The standardized curves are generated from HRM curves from a plurality of samples, each sample having a known but different methylation status. In one embodiment, the first negative derivative of each HRM curve from the known samples is plotted and a first value corresponding with a first melt peak and a second value corresponding with a second melt peak from the negative derivative plots are identified. The slope of a line connecting the first and second values for each sample is calculated and used to identify a slope data point that is plotted to generate the standardized curve. In another embodiment, a threshold line that intersects the plurality of HRM curves is generated and the standardized curve is generated from the intersection data points.
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