Elsevier

Analytical Biochemistry

Volume 379, Issue 1, 1 August 2008, Pages 127-129
Analytical Biochemistry

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Standardization of real-time PCR gene expression data from independent biological replicates

https://doi.org/10.1016/j.ab.2008.04.036Get rights and content

Abstract

Gene expression analysis by quantitative reverse transcription PCR (qRT–PCR) allows accurate quantifications of messenger RNA (mRNA) levels over different samples. Corrective methods for different steps in the qRT–PCR reaction have been reported; however, statistical analysis and presentation of substantially variable biological repeats present problems and are often not meaningful, for example, in a biological system such as mouse embryonic stem cell differentiation. Based on a series of sequential corrections, including log transformation, mean centering, and autoscaling, we describe a robust and powerful standardization method that can be used on highly variable data sets to draw statistically reliable conclusions.

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Acknowledgments

This research was funded by the Horizontal Action of the Vrije Universiteit Brussel (grant HOA1), Fund for Scientific Research Flanders (grant G.0485.06), and the European Union (Sixth Framework, grant NEST012930). Jo Vandesompele is supported by the Fund for Scientific Research Flanders (FWO).

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