PCR-Based Determination of Mitochondrial DNA Copy Number in Multiple Species

Methods Mol Biol. 2021:2310:91-111. doi: 10.1007/978-1-0716-1433-4_8.

Abstract

Mitochondrial DNA (mtDNA) copy number is a critical component of overall mitochondrial health. In this chapter, we describe methods for simultaneous isolation of mtDNA and nuclear DNA (nucDNA), and measurement of their respective copy numbers using quantitative PCR. Methods differ depending on the species and cell type of the starting material, and availability of specific PCR reagents. We also briefly describe factors that affect mtDNA copy number and discuss caveats to its use as a biomarker.

Keywords: Copy number; Mitochondrial DNA; Mitochondrial disease; Mitochondrial toxicity; QPCR; mtDNA; mtDNA depletion.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Caenorhabditis elegans / genetics
  • Cell Nucleus / genetics*
  • DNA Copy Number Variations*
  • DNA, Fungal / genetics
  • DNA, Mitochondrial / genetics*
  • Drosophila melanogaster / genetics
  • Fundulidae / genetics
  • Gene Dosage*
  • Humans
  • Mice
  • Mitochondria / genetics*
  • Oryzias / genetics
  • Rats
  • Real-Time Polymerase Chain Reaction*
  • Saccharomyces cerevisiae / genetics
  • Zebrafish / genetics

Substances

  • DNA, Fungal
  • DNA, Mitochondrial