Počet záznamů: 1

Statistical aspects of quantitative real-time PCR experiment design

  1. 1.
    0350829 - BTO-N 2011 RIV US eng J - Článek v odborném periodiku
    Kitchen, R.R. - Kubista, Mikael - Tichopád, Aleš
    Statistical aspects of quantitative real-time PCR experiment design.
    Methods. Roč. 50, č. 4 (2010), s. 231-236 ISSN 1046-2023
    Grant CEP: GA AV ČR IAA500520809
    Výzkumný záměr: CEZ:AV0Z50520701
    Klíčová slova: Real-time PCR * Experiment design * Nested analysis of variance
    Kód oboru RIV: EB - Genetika a molekulární biologie
    Impakt faktor: 4.527, rok: 2010

    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
    Trvalý link: http://hdl.handle.net/11104/0190725