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Analyzing real-time PCR data by the comparative C(T) method.

Nature protocols

Animals, genetics, Transcription Factors, RNA, Messenger, RNA, statistics & numerical data, methods, Polymerase Chain Reaction, Pancreatic Neoplasms, metabolism, Pancreas, MicroRNAs, Mice, Humans, Homeodomain Proteins, Genes, bcl-2, Gene Expression Profiling, Gene Expression, Female, Data Interpretation, Statistical, Computer Systems, Breast Neoplasms

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      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 (-DeltaDeltaC(T)) 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.

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