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The integrity and assay performance of tissue mitochondrial DNA is considerably affected by choice of isolation method
Umeå University, Faculty of Medicine, Department of Medical Biochemistry and Biophysics.ORCID iD: 0000-0001-5704-3093
Umeå University, Faculty of Medicine, Department of Medical Biochemistry and Biophysics.
Umeå University, Faculty of Medicine, Department of Medical Biochemistry and Biophysics.
Umeå University, Faculty of Medicine, Department of Medical Biochemistry and Biophysics.
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2021 (English)In: Mitochondrion (Amsterdam. Print), ISSN 1567-7249, E-ISSN 1872-8278, Vol. 61, p. 179-187Article in journal (Refereed) Published
Abstract [en]

The integrity of mitochondrial DNA (mtDNA) isolated from solid tissues is critical for analyses such as long-range PCR, but is typically assessed under conditions that fail to provide information on the individual mtDNA strands. Using denaturing gel electrophoresis, we show that commonly-used isolation procedures generate mtDNA containing several single-strand breaks per strand. Through systematic comparison of DNA isolation methods, we identify a procedure yielding the highest integrity of mtDNA that we demonstrate displays improved performance in downstream assays. Our results highlight the importance of isolation method choice, and serve as a resource to researchers requiring high-quality mtDNA from solid tissues.

Place, publisher, year, edition, pages
Elsevier, 2021. Vol. 61, p. 179-187
Keywords [en]
DNA integrity, Long-range PCR, Mitochondrial DNA, mtDNA, Nuclease activity
National Category
Cell and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-189552DOI: 10.1016/j.mito.2021.10.005ISI: 000717836700001Scopus ID: 2-s2.0-85118478892OAI: oai:DiVA.org:umu-189552DiVA, id: diva2:1611754
Funder
Åke Wiberg Foundation, M20-0132Swedish Cancer Society, 19 0022 JIA, 190098 Pj 01 HSwedish Society for Medical Research (SSMF), S17-0023Swedish Research Council, 2019-01874Available from: 2021-11-16 Created: 2021-11-16 Last updated: 2023-09-05Bibliographically approved

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Repolês, Bruno MarçalGorospe, Choco MichaelTran, PhongNilsson, Anna KarinWanrooij, Paulina H.

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Repolês, Bruno MarçalGorospe, Choco MichaelTran, PhongNilsson, Anna KarinWanrooij, Paulina H.
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Department of Medical Biochemistry and Biophysics
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Mitochondrion (Amsterdam. Print)
Cell and Molecular Biology

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