title: Multiplex Analyses Using Real-Time Quantitative PCR.
abstract: Quantitative polymerase chain reaction (qPCR) is a routinely used method for the detection and quantitation of gene expression in real time. Multiplex qPCR requires the use of probe-based assays, in which each probe is labeled with a unique fluorescent dye, resulting in different observed colors for each assay. The signal from each dye is used to quantitate the amount of each target separately in the same tube or well. The availability to multiplex therefore allows the measurement of the expression levels of several targets or genes of interest quickly. Here, we describe a method using the SensiFAST and SensiFAST One-Step probe kits which allows simultaneous real-time quantitation of up to 5 amplicons.
authors.affiliation: Bioline Reagents Limited, Unit 16, The Edge Business Centre, Humber Road, London, NW2 6EW, UK. steve.hawkins@bioline.com.
authors.forename: Steve F C
authors.initials: SFC
authors.lastname: Hawkins
authors.affiliation: Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Campinas, Brazil.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Guest
mesh_terms: D015415:Biomarkers; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods / Q000592:standards; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D015415:Biomarkers
keywords: Amplicon; Fluorescent dyes; Multiplex analysis; Quantitation; Taq polymerase; cDNA; mRNA; qPCR