About: Statistical aspects of quantitative real-time PCR experiment design

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An Entity of Type : http://linked.opendata.cz/ontology/domain/vavai/Vysledek, within Data Space : linked.opendata.cz associated with source document(s)

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rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
Description	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 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 (en)
Title	Statistical aspects of quantitative real-time PCR experiment design Statistical aspects of quantitative real-time PCR experiment design (en)
skos:prefLabel	Statistical aspects of quantitative real-time PCR experiment design Statistical aspects of quantitative real-time PCR experiment design (en)
skos:notation	RIV/86652036:_____/10:00350829!RIV11-AV0-86652036
http://linked.open...avai/riv/aktivita	P Z
http://linked.open...avai/riv/aktivity	P(IAA500520809), Z(AV0Z50520701)
http://linked.open...iv/cisloPeriodika	4
http://linked.open...vai/riv/dodaniDat	2011
http://linked.open...aciTvurceVysledku	Kubista, Mikael Tichopád, Aleš
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Biotechnologický ústav AV ČR, v. v. i.
http://linked.open...dnocenehoVysledku	289933
http://linked.open...ai/riv/idVysledku	RIV/86652036:_____/10:00350829
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	Real-time PCR; Experiment design; Nested analysis of variance (en)
http://linked.open.../riv/klicoveSlovo	Real-time PCR Experiment design Nested analysis of variance
http://linked.open...odStatuVydavatele	US - Spojené státy americké
http://linked.open...ontrolniKodProRIV	[FAD99BE743E5]
http://linked.open...i/riv/nazevZdroje	Methods
http://linked.open...in/vavai/riv/obor	EB
http://linked.open...ichTvurcuVysledku	2 (xsd:int)
http://linked.open...cetTvurcuVysledku	3 (xsd:int)
http://linked.open...vavai/riv/projekt	Mechanisms of embryonic stem cells early differentiation
http://linked.open...UplatneniVysledku	2010
http://linked.open...v/svazekPeriodika	50
http://linked.open...iv/tvurceVysledku	Kubista, Mikael Tichopád, Aleš Kitchen, R. R.
http://linked.open...ain/vavai/riv/wos	000276262100004
http://linked.open...n/vavai/riv/zamer	Establishing of the Biotechnology Institute ASCR
issn	1046-2023
number of pages	6 (xsd:int)

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