Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCT method  (Article)

Livak, K.J.^a, Schmittgen, T.D.^b  

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^a  Applied Biosystems, Foster City, CA 94404, United States
^b  Department of Pharmaceutical Sciences, College of Pharmacy, Washington State University, Pullman, WA 99164-6534, United States

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Abstract

The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2^-ΔΔCT method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2^-ΔΔCT method. In addition, we present the derivation and applications of two variations of the 2^-ΔΔCT method that may be useful in the analysis of real-time, quantitative PCR data. © 2001 Elsevier Science (USA).

Author keywords

Quantitative polymerase chain reaction; Real-time polymerase chain reaction; Relative quantification; Reverse transcription polymerase chain reaction; Taq Man.

Indexed keywords

EMTREE drug terms: complementary DNA

EMTREE medical terms: accuracy; article; copy number; data analysis; diagnostic value; gene duplication; gene expression regulation; intermethod comparison; mathematical computing; polymerase chain reaction; priority journal; process design; quantitative diagnosis; real time quantitative polymerase chain reaction; signal detection; statistical analysis; validation process

MeSH: Algorithms; Brain; Cell Line; DNA, Complementary; Humans; Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Time Factors
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Chemicals and CAS Registry Numbers: DNA, Complementary