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CSB
2005
IEEE

Automated Validation of Polymerase Chain Reactions Using Amplicon Melting Curves

14 years 25 days ago
Automated Validation of Polymerase Chain Reactions Using Amplicon Melting Curves
PCR, the polymerase chain reaction, is a fundamental tool of molecular biology. Quantitative PCR is the goldstandard methodology for determination of DNA copy numbers, quantitating transcription, and numerous other applications. A major barrier to large-scale application of PCR for quantitative genomic analyses is the current requirement for manual validation of individual PCR reactions to ensure generation of a single product. This typically requires visual inspection either of gel electrophoreses or temperature dissociation (“melting”) curves of individual PCR reactions—a time-consuming and costly process. Here we describe a robust computational solution to this fundamental problem. Using a training set of 10,080 reactions comprising multiple quantitative PCR reactions from each of 1,728 unique human genomic amplicons, we developed a support vector machine classifier capable of discriminating single-product PCR reactions with better than 99% accuracy. This approach has broad ...
Tobias P. Mann, Richard Humbert, John A. Stamatoya
Added 24 Jun 2010
Updated 24 Jun 2010
Type Conference
Year 2005
Where CSB
Authors Tobias P. Mann, Richard Humbert, John A. Stamatoyannopoulos, William Stafford Noble
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