Počet záznamů: 1

Design and Optimization of Reverse-Transcription Quantitative PCR Experiments

  1. 1.
    0339069 - BTO-N 2010 RIV US eng J - Článek v odborném periodiku
    Tichopád, A. - Kitchen, R. - Riedmaier, I. - Becker, Ch. - Ståhlberg, A. - Kubista, Mikael
    Design and Optimization of Reverse-Transcription Quantitative PCR Experiments.
    Clinical Chemistry. Roč. 55, č. 10 (2009), s. 1816-1823 ISSN 0009-9147
    Výzkumný záměr: CEZ:AV0Z50520701
    Klíčová slova: Design * optimization * RT qPCR
    Kód oboru RIV: EG - Zoologie
    Impakt faktor: 6.263, rok: 2009

    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
    Trvalý link: http://hdl.handle.net/11104/0182679