Analyzing real-time PCR data by the comparative CT method

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Schmittgen, Thomas D.¹; Livak, Kenneth J.²

Analyzing real-time PCR data by the comparative C_T method

Nat Protoc. 2008;3(6):1101-1108

©2008 Nature Publishing Group

Affiliations

¹Division of Pharmaceutics, College of Pharmacy, Ohio State University, Parks Hall, 500 West 12th Avenue, Columbus, Ohio, OH 43210 USA

²Applied Biosystems, 850 Lincoln Center Drive, Foster City, California, CA 94404 USA
Abstract

Two different methods of presenting quantitative gene expression exist: absolute and relative quantification. Absolute quantification calculates the copy number of the gene usually by relating the PCR signal to a standard curve. Relative gene expression presents the data of the gene of interest relative to some calibrator or internal control gene. A widely used method to present relative gene expression is the comparative C_T method also referred to as the 2^-ΔΔ𝐶T method. This protocol provides an overview of the comparative C_T method for quantitative gene expression studies. Also presented here are various examples to present quantitative gene expression data using this method.
Links

DOI: 10.1038/nprot.2008.73

PubMed: 18546601

WoS: 000258423700018
History

Online: 2008-06-05

Cited Works (2)

Year	Entry
2001	Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods. December 2001;25(4):402-408.
2001	Pfaffl MW. A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res. May 1, 2001;29(9):e45.

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