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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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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