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Quality control for quantitative PCR based on amplification compatibility test
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SYSNO ASEP 0351162 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Quality control for quantitative PCR based on amplification compatibility test Author(s) Tichopád, Aleš (BTO-N)
Bar, T. (IL)
Pecen, Ladislav (BTO-N)
Kitchen, R.R. (GB)
Kubista, Mikael (BTO-N) RID
Pfaffl, M.W. (DE)Source Title Methods. - : Elsevier - ISSN 1046-2023
Roč. 50, č. 4 (2010), s. 308-312Number of pages 5 s. Language eng - English Country US - United States Keywords Quantitative PCR ; Quality control ; Amplification efficiency Subject RIV EB - Genetics ; Molecular Biology R&D Projects IAA500520809 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) IAA500970904 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) CEZ AV0Z50520701 - BTO-N (2007-2013) UT WOS 000276262100015 DOI 10.1016/j.ymeth.2010.01.028 Annotation Quantitative qPCR is method for the accurate quantification of nucleic acids. Yet it may generate erroneous results if the amplification process is obscured by inhibition or generation of aberrant side-product. Herein we present a statistical approach based on multivariate analysis of the amplification response data generated in qPCR. The amplification trajectory in its most dynamic phase is fitted with a suitable model. Two parameters of this model, related to amplification efficiency, are then used for calculation of the Z-score statistics. Each studied sample is compared to a predefined reference set of reactions. A probabilistic decision for each individual Z-score is then used to identify the majority of inhibited reactions in our experiments. We demonstrate improved identification performance using the multivariate approach Finally we stress that the performance of the amplification kompatibility test as a quality control procedure depends on the quality of the reference set Workplace Institute of Biotechnology Contact Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Year of Publishing 2011
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