Droplet Digital PCR with EvaGreen Assay: Confirmational Analysis of Structural Variants

Curr Protoc Hum Genet. 2018 Apr;97(1):e58. doi: 10.1002/cphg.58.

Abstract

DNA structural variants can be analyzed by droplet digital PCR (ddPCR), a water-oil microfluidics and fluorescence technology to quantify target nucleic acids with extreme precision and sensitivity. Traditional ddPCR uses expensive fluorescent oligonucleotide probes that require extensive optimization. Here we describe a variation of ddPCR using a DNA-binding dye (EvaGreen), whose properties allow target products to be effectively quantified at a significantly lower cost. © 2018 by John Wiley & Sons, Inc.

Keywords: EvaGreen; copy number variant; ddPCR; rare variant detections; structural variant.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay
  • DNA / analysis*
  • Humans
  • Molecular Probes / chemistry*
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Single Nucleotide*

Substances

  • Molecular Probes
  • DNA