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Statistical aspects of quantitative real-time PCR experiment design
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SYSNO ASEP 0350829 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Statistical aspects of quantitative real-time PCR experiment design Author(s) Kitchen, R.R. (GB)
Kubista, Mikael (BTO-N) RID
Tichopád, Aleš (BTO-N)Source Title Methods. - : Elsevier - ISSN 1046-2023
Roč. 50, č. 4 (2010), s. 231-236Number of pages 6 s. Language eng - English Country US - United States Keywords Real-time PCR ; Experiment design ; Nested analysis of variance Subject RIV EB - Genetics ; Molecular Biology R&D Projects IAA500520809 GA AV ČR - Academy of Sciences of the Czech Republic (AV ČR) CEZ AV0Z50520701 - BTO-N (2007-2013) UT WOS 000276262100004 DOI 10.1016/j.ymeth.2010.01.025 Annotation Experiments using qPCR to test hypotheses are limited by technical and biological variability; we seek to minimise sources of variability through optimum use of biological and technical replicates. Calculations rely on knowledge of the expected variances of the measurements of each group of samples and the magnitude of the treatment effect. Here we introduce a method that exploits a small pilot study to estimate the biological and technical variances in order to improve the design of a subsequent large experiment. We measure the variance contributions at several ‘levels’ of the experiment design and provide a means of using this information to predict both the total variance and the prospective power of the assay. We also discuss the effect of normalisation to a reference gene in terms of the measured variance components of the gene of interest. Finally, we describe a software implementation of these methods, powerNest Workplace Institute of Biotechnology Contact Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Year of Publishing 2011
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