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BMCBI
2010

Shape based kinetic outlier detection in real-time PCR

10 years 2 months ago
Shape based kinetic outlier detection in real-time PCR
Background: Real-time PCR has recently become the technique of choice for absolute and relative nucleic acid quantification. The gold standard quantification method in real-time PCR assumes that the compared samples have similar PCR efficiency. However, many factors present in biological samples affect PCR kinetic, confounding quantification analysis. In this work we propose a new strategy to detect outlier samples, called SOD. Results: Richards function was fitted on fluorescence readings to parameterize the amplification curves. There was not a significant correlation between calculated amplification parameters (plateau, slope and y-coordinate of the inflection point) and the Log of input DNA demonstrating that this approach can be used to achieve a "fingerprint" for each amplification curve. To identify the outlier runs, the calculated parameters of each unknown sample were compared to those of the standard samples. When a significant underestimation of starting DNA molec...
Davide Sisti, Michele Guescini, Marco B. L. Rocchi
Added 08 Dec 2010
Updated 08 Dec 2010
Type Journal
Year 2010
Where BMCBI
Authors Davide Sisti, Michele Guescini, Marco B. L. Rocchi, Pasquale Tibollo, Mario D'Atri, Vilberto Stocchi
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