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Loop-Mediated Isothermal Amplification for the Detection of Plant Pathogens

  • Lisa I. WardEmail author
  • Scott J. Harper
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 862)

Abstract

Loop-mediated isothermal amplification (LAMP) is a technique involving the use of four to six primers (two inner primers, two outer primers, and two loop primers) and the strand displacement activity of Bacillus subtilis-derived (Bst) DNA polymerase. The end result of strand displacement and loop formation and synthesis is the single-temperature amplification of a highly specific fragment from a DNA template at a much greater titre than that obtained with polymerase chain reaction. With LAMP, there are several methods to determine a positive reaction. Presented here are three alternative methods: gel electrophoresis, hydroxynaphthol blue colorimetric dye, and the fluorescent intercalating PicoGreen® reagent.

Key words

Loop-mediated isothermal amplification Strand displacement End point detection Hydroxynaphthol blue Picogreen Gel electrophoresis 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Plant Health and Environment Laboratory, Investigation and Diagnostic CentreMAF Biosecurity New ZealandWellingtonNew Zealand
  2. 2.Citrus Research and Education CenterLaboratory University of FloridaLake AlfredUSA

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