title: Multiplex Single Nucleotide Polymorphism Analyses.
abstract: Quantitative polymerase chain reaction (qPCR) is a routinely used method for detection and quantitation of gene expression in real time. This is achieved through the incorporation and measurement of fluorescent reporter probes in the amplified cDNA strands, since the fluorescent signals increase as the reaction progresses. The availability of multiple probes that fluoresce at different wavelengths allows for multiplexing as this gives rise to amplicons with unique fluorescent signatures. 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. shawkins@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: D000483:Alleles; D019295:Computational Biology / Q000379:methods; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012984:Software; D013223:Statistics as Topic
keywords: Allele; Fluorescent dyes; Multiplex analysis; PCR; Quantitation; Single nucleotide polymorphism; Taq polymerase