title: Explanatory chapter: quantitative PCR.
abstract: Quantitative PCR (qPCR), also called real-time PCR or quantitative real-time PCR, is a PCR-based technique that couples amplification of a target DNA sequence with quantification of the concentration of that DNA species in the reaction. This method enables calculation of the starting template concentration and is therefore a frequently used analytical tool in evaluating DNA copy number, viral load, SNP detection, and allelic discrimination. When preceded by reverse-transcription PCR, qPCR is a powerful tool to measure mRNA expression and is the gold standard for microarray gene expression data confirmation. Given the broad applications of qPCR and the many technical variations that have been developed, a brief survey of qPCR, including technical background, available chemistries, and data analysis techniques will provide a framework for both experimental design and evaluation.
authors.affiliation: The High Throughput Biology Center and Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: jsiege16@jhmi.edu.
authors.forename: Jessica S
authors.initials: JS
authors.lastname: Dymond
mesh_terms: D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D056915:DNA Copy Number Variations / Q000235:genetics; D005456:Fluorescent Dyes; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D019562:Viral Load / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D004247:DNA
keywords: Amplification efficiency; Primer design; Quantitative PCR; SYBR Green I (SG); TaqMan Probes