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Optimized loop-mediated isothermal amplification (lamp) allows single copy detection using bioluminescent assay in real time (bart)

Hardinge, Patrick ORCID: https://orcid.org/0000-0002-6292-5186 2022. Optimized loop-mediated isothermal amplification (lamp) allows single copy detection using bioluminescent assay in real time (bart). Bioluminescence: Methods and Protocols, Vol. 2524. Springer, pp. 107-117. (10.1007/978-1-0716-2453-1_8)

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Abstract

The bioluminescent assay in real time (BART) is a molecular diagnostic tool for the detection of nucleic acid amplification by recording light output. The key component for BART is a thermostable luciferase derived from the firefly Photinus pyralis. Coupling BART detection with the isothermal amplification method loopmediated isothermal amplification (LAMP) enables rapid molecular diagnostic results using simple equipment. LAMP-BART provides quantitative results from a closed tube and is appropriate to microliter standard tests and nanoliter microfluidic assays. In this chapter, we introduce a protocol to amplify and detect genetic markers using LAMP with BART. Furthermore, we provide advice to optimize LAMP assays for high sensitivity and specificity and to eliminate the incidence of “false positive” results which can occur from the components of the assay. The optimization of genetically modified (GM) maize by targeting the nopaline synthase terminator (NOSt) and 35S promoter (35Sp) sequences is described

Item Type: Book Section
Date Type: Published Online
Status: Published
Schools: Biosciences
Publisher: Springer
ISBN: 978-1-0716-2453-1
Last Modified: 03 Nov 2022 14:45
URI: https://orca.cardiff.ac.uk/id/eprint/153291

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