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Channels (Austin). 2013 May-Jun;7(3):160-70. doi: 10.4161/chan.24024. Epub 2013 Mar 19.

Efficient experimental design and analysis of real-time PCR assays.

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  • 1Department of Physiology, Faculty of Medicine, University of Toronto, Toronto, ON, Canada.

Abstract

Real-time polymerase chain reaction (qPCR) is currently the standard for gene quantification studies and has been extensively used in large-scale basic and clinical research. The operational costs and technical errors can become a significant issue due to the large number of sample reactions. In this paper, we present an experimental design strategy and an analysis procedure that are more efficient requiring fewer sample reactions than the traditional approach. We verified mathematically and experimentally the new design on a well-characterized model, to evaluate the gene expression levels of CACNA1C and CACNA1G in hypertrophic ventricular myocytes induced by phenylephrine treatment.

KEYWORDS:

ANF; L-type calcium channel; Real-time PCR; T-type calcium channel; cell culture; gene expression; hypertrophy; ventricular myocytes

PMID:
23510941
[PubMed - indexed for MEDLINE]
PMCID:
PMC3710343
Free PMC Article
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