title: Standardization of real-time PCR gene expression data from independent biological replicates.
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.
authors.affiliation: Del E. Webb Neuroscience, Aging, and Stem Cell Research Center, Burnham Institute for Medical Research, La Jolla, CA 92037, USA. ewillems@burnham.org
authors.forename: Erik
authors.initials: E
authors.lastname: Willems
authors.forename: Luc
authors.initials: L
authors.lastname: Leyns
authors.forename: Jo
authors.initials: J
authors.lastname: Vandesompele
mesh_terms: D000818:Animals; D053595:Embryonic Stem Cells / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics*; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards
chemical_list: D012333:RNA, Messenger