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PCR Based Determination of Mitochondrial DNA Copy Number in Multiple Species

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Mitochondrial Regulation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1241))

Abstract

Mitochondrial DNA (mtDNA) copy number is a critical component of overall mitochondrial health. In this chapter, we describe methods for isolation of both 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.

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Acknowledgements

We thank Aleksandra Trifunovic for sharing the mtDNA copy number plasmid for C. elegans with us, and we thank Kevin Kwok for supplying us with medaka tissue. This work was supported by P42 ES010356-10A2, and the National Science Foundation (NSF) and the Environmental Protection Agency under NSF Cooperative Agreement EF-0830093, Center for the Environmental Implications of NanoTechnology (CEINT). Any opinions, findings, conclusions, or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the NSF or the EPA. This work has not been subjected to EPA review and no official endorsement should be inferred. The funding sources had no role in experimental design, data collection, or interpretation.

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Authors and Affiliations

  1. Nicholas School of the Environment, Duke University, 450 Research Dr., Durham, NC, 27708-0328, USA

    John P. Rooney, Ian T. Ryde, Meryl D. Colton & Joel N. Meyer Ph.D.

  2. Department of Neurology, Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, PA, 15260, USA

    Laurie H. Sanders, Evan. H. Howlett & J. Timothy Greenamyre

  3. The Institute of Environmental and Human Health, Texas Tech University, Lubbock, TX, 79401, USA

    Kaylyn E. Germ & Greg D. Mayer

Authors
  1. John P. Rooney
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  2. Ian T. Ryde
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  3. Laurie H. Sanders
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  4. Evan. H. Howlett
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  5. Meryl D. Colton
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  6. Kaylyn E. Germ
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  7. Greg D. Mayer
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  8. J. Timothy Greenamyre
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  9. Joel N. Meyer Ph.D.
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Corresponding author

Correspondence to Joel N. Meyer Ph.D. .

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Editors and Affiliations

  1. Department of Life Sciences and Center for Neurosciences and Cell Biology, University of Coimbra, Coimbra, Portugal

    Carlos M. Palmeira

  2. Department of Biology and Center for Neurosciences and Cell Biology, University of Aveiro and University of Coimbra, Aveiro and Coimbra, Portugal

    Anabela P. Rolo

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Rooney, J.P. et al. (2015). PCR Based Determination of Mitochondrial DNA Copy Number in Multiple Species. In: Palmeira, C., Rolo, A. (eds) Mitochondrial Regulation. Methods in Molecular Biology, vol 1241. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1875-1_3

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  • DOI: https://doi.org/10.1007/978-1-4939-1875-1_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1874-4

  • Online ISBN: 978-1-4939-1875-1

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