Počet záznamů: 1
Design and Optimization of Reverse-Transcription Quantitative PCR Experiments
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SYSNO ASEP 0339069 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Design and Optimization of Reverse-Transcription Quantitative PCR Experiments Tvůrce(i) Tichopád, A. (DE)
Kitchen, R. (GB)
Riedmaier, I. (DE)
Becker, Ch. (DE)
Ståhlberg, A. (SE)
Kubista, Mikael (BTO-N) RIDZdroj.dok. Clinical Chemistry. - : Oxford University Press - ISSN 0009-9147
Roč. 55, č. 10 (2009), s. 1816-1823Poč.str. 8 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova Design ; optimization ; RT qPCR Vědní obor RIV EG - Zoologie CEZ AV0Z50520701 - BTO-N (2007-2013) UT WOS 000270471300010 DOI 10.1373/clinchem.2009.126201 Anotace Quantitative PCR (qPCR) is a valuable technique for accurately and reliably profiling and quantifying gene expression. Typically, samples obtained from the organism of study have to be processed via several preparative steps before qPCR. We estimated the errors of sample withdrawal and extraction, reverse transcription (RT), and qPCR that are introduced into measurements of mRNA concentrations. We performed hierarchically arranged experiments with 3 animals, 3 samples, 3 RT reactions, and 3 qPCRs and quantified the expression of several genes in solid tissue, blood, cell culture, and single cells.A nested ANOVA design was used to model the experiments, and relative and absolute errors were calculated with this model for each processing level in the hierarchical design. We recommend the use of sample replicates preferentially to any other replicates when working with solid tissue, cell cultures,and single cells,and we recommend the use of RT replicates when working with blood Pracoviště Biotechnologický ústav Kontakt Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Rok sběru 2010
Počet záznamů: 1