Abstract
High specificity has been demonstrated in polymerase chain reaction (PCR) with the use of molecular beacons (MBs) to detect amplified sequences containing mutations or single-nucleotide polymorphisms (SNPs). MBs have been adapted for use with the isothermal nucleic acid amplification technology loop-mediated amplification (LAMP) by targeting single-stranded loop sequences under optimized conditions to enable applications such as plant genotyping. LAMP has several benefits over PCR, such as rapid amplification, single-temperature reaction conditions enabling low-cost equipment, and robustness to certain PCR inhibitors. However, and despite the increased number of primers required, the specificity of LAMP is limited, and false positive results can be problematic. In this chapter, design considerations for molecular beacons in LAMP assays are described, as well as a method for MB-LAMP amplification and detection, with an example of gene sequences in genetically modified (GM) maize samples.
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Acknowledgments
The research into molecular beacons in LAMP assays was funded by BBSRC Stand Alone LINK grant BB/L022346/1, a BBSRC Industrial CASE award BB/G017336/1, and the European Regional Development Fund with Cardiff University 80762-CU-085 (East).
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Hardinge, P. (2023). Molecular Beacons – Loop-Mediated Amplification (MB-LAMP). In: Shavrukov, Y. (eds) Plant Genotyping. Methods in Molecular Biology, vol 2638. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3024-2_20
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DOI: https://doi.org/10.1007/978-1-0716-3024-2_20
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