Quantitative Analysis with Droplet Digital PCR

Methods Mol Biol. 2019:1875:171-186. doi: 10.1007/978-1-4939-8837-2_14.

Abstract

Digital PCR-based methods, such as droplet digital PCR, are one of the best tools for determination of absolute nucleic-acid copy numbers. These techniques avoid the need for reference materials with known target concentrations. Compared to real-time PCR, they provide higher accuracy of quantification at low target concentrations, and have higher resilience to inhibitors. In this Chapter, we describe the droplet digital PCR workflow for the detection and quantification of flavescence dorée phytoplasma.

Keywords: Absolute quantification; Digital PCR; Droplet digital PCR; Flavescence dorée phytoplasma; Phytoplasma reference material.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Computer Simulation
  • DNA, Bacterial / analysis*
  • Phytoplasma / genetics
  • Phytoplasma / isolation & purification*
  • Plant Diseases / microbiology
  • Real-Time Polymerase Chain Reaction / methods*
  • Vitis / microbiology*
  • Workflow

Substances

  • DNA, Bacterial