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Multiplex Analyses Using Real-Time Quantitative PCR

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Multiplex Biomarker Techniques

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1546))

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.

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Correspondence to Steve F. C. Hawkins .

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Hawkins, S.F.C., Guest, P.C. (2017). Multiplex Analyses Using Real-Time Quantitative PCR. In: Guest, P.C. (eds) Multiplex Biomarker Techniques. Methods in Molecular Biology, vol 1546. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-6730-8_8

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  • DOI: https://doi.org/10.1007/978-1-4939-6730-8_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-6729-2

  • Online ISBN: 978-1-4939-6730-8

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