Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2(-Delta Delta C(T)) Method

Methods. 2001 Dec;25(4):402-8. doi: 10.1006/meth.2001.1262.

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(-Delta Delta C(T)) 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(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data.

MeSH terms

  • Algorithms
  • Brain / metabolism
  • Cell Line
  • DNA, Complementary / metabolism
  • Humans
  • Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Time Factors

Substances

  • DNA, Complementary