Analysis of relative gene expression data using real-time quantitative

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Analysis of relative gene expression data using real-time quantitative (2001)

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by Kenneth J. Livak , Thomas D. Schmittgen

Venue:	PCR and 2 ��CT method. Methods 25
Citations:	2664 - 6 self

BibTeX

@INPROCEEDINGS{Livak01analysisof,
    author = {Kenneth J. Livak and Thomas D. Schmittgen},
    title = {Analysis of relative gene expression data using real-time quantitative},
    booktitle = {PCR and 2 ��CT method. Methods 25},
    year = {2001},
    pages = {402--408}
}

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Abstract

of the target gene relative to some reference group The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantifica-such as an untreated control or a sample at time zero tion and relative quantification. Absolute quantification deter- in a time-course study. mines the input copy number, usually by relating the PCR signal Absolute quantification should be performed in situ-to a standard curve. Relative quantification relates the PCR signal ations where it is necessary to determine the absolute of the target transcript in a treatment group to that of another transcript copy number. Absolute quantification has sample such as an untreated control. The 2��CT method is a been combined with real-time PCR and numerous reconvenient way to analyze the relative changes in gene expression ports have appeared in the literature (6–9) including from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications two articles in this issue (10, 11). In some situations, of the 2 it may be unnecessary to determine the absolute tran-method. In addition, we present the derivation and applications of two variations of the 2 ��CT method that may be script copy number and reporting the relative change useful in the analysis of real-time, quantitative PCR data. � 2001 in gene expression will suffice. For example, stating

Keyphrases

real-time quantitative relative gene expression data relative change relative quantification ct method untreated control gene expression standard curve pcr signal ations absolute quantification deter pcr signal absolute quantification gene expression port real-time quantitative pcr experiment treatment group numerous reconvenient way quantitative pcr data quantitative pcr experiment real-time pcr absolute quantification reference group absolute tran-method time-course study target transcript transcript copy number input copy number target gene script copy number

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