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Statistical aspects of quantitative real-time PCR experiment design
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SYSNO ASEP 0350829 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Statistical aspects of quantitative real-time PCR experiment design Tvůrce(i) Kitchen, R.R. (GB)
Kubista, Mikael (BTO-N) RID
Tichopád, Aleš (BTO-N)Zdroj.dok. Methods. - : Elsevier - ISSN 1046-2023
Roč. 50, č. 4 (2010), s. 231-236Poč.str. 6 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova Real-time PCR ; Experiment design ; Nested analysis of variance Vědní obor RIV EB - Genetika a molekulární biologie CEP IAA500520809 GA AV ČR - Akademie věd CEZ AV0Z50520701 - BTO-N (2007-2013) UT WOS 000276262100004 DOI 10.1016/j.ymeth.2010.01.025 Anotace 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 Pracoviště Biotechnologický ústav Kontakt Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Rok sběru 2011
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