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Curr Protoc Toxicol. 2019 Apr 14:e75. doi: 10.1002/cptx.75. [Epub ahead of print]

Analysis of Human Mitochondrial DNA Content by Southern Blotting and Nonradioactive Probe Hybridization.

Author information

1
Department of Biochemistry and Molecular Biology, Southern Illinois University School of Medicine, Carbondale, Illinois.

Abstract

A single cell can contain several thousand copies of the mitochondrial DNA genome or mtDNA. Tools for assessing mtDNA content are necessary for clinical and toxicological research, as mtDNA depletion is linked to genetic disease and drug toxicity. For instance, mtDNA depletion syndromes are typically fatal childhood disorders that are characterized by severe declines in mtDNA content in affected tissues. Mitochondrial toxicity and mtDNA depletion have also been reported in human immunodeficiency virus-infected patients treated with certain nucleoside reverse transcriptase inhibitors. Further, cell culture studies have demonstrated that exposure to oxidative stress stimulates mtDNA degradation. Here we outline a Southern blot and nonradioactive digoxigenin-labeled probe hybridization method to estimate mtDNA content in human genomic DNA samples.

KEYWORDS:

Drug-induced mitochondrial toxicity; Southern blotting; digoxigenin-labeling of DNA probes; immunodetection; mitochondrial DNA (mtDNA) depletion

PMID:
30982231
DOI:
10.1002/cptx.75

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