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Analysis of Relative Gene Expression Data Using …
METHODS 25, 402 408 (2001). , available online at on Analysis of Relative gene Expression data Using Real- Time Quantitative PCR and the 2 CT Method Kenneth J. Livak* and Thomas D. Schmittgen ,1. *Applied Biosystems, Foster City, California 94404; and Department of Pharmaceutical Sciences, College of Pharmacy, Washington State University, Pullman, Washington 99164-6534. 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.
404 LIVAK AND SCHMITTGEN results of an experiment where a cDNA preparation change in gene expression relative to an untreated con-was diluted over a 100-fold range.
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