The DOI matches its Crossref record.
Quantitative PCR-Based Measurement of Nuclear and Mitochondrial DNA Damage and Repair in Mammalian Cells
Abstract
In this chapter, we describe a gene-specific quantitative polymerase chain reaction (QPCR)-based assay for the measurement of DNA damage, using amplification of long DNA targets. This assay has been extensively used to measure the integrity of both nuclear and mitochondrial genomes exposed to different genotoxins, and has proved particularly valuable in identifying reactive oxygen species-mediated mitochondrial DNA (mtDNA) damage. QPCR can be used to quantify the formation of DNA damage, as well as the kinetics of damage removal. One of the main strengths of the assay is that it permits monitoring the integrity of mtDNA directly from total cellular DNA without the need for isolating mitochondria, or a separate step of mtDNA purification. Here we discuss advantages and limitations of using QPCR to assay DNA damage in mammalian cells. In addition, we give a detailed protocol for the QPCR assay that helps facilitate its successful deployment in any molecular biology laboratory.
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Cited by
Gomes, A. P., Price, N. L., Ling, A. J. Y., Moslehi, J. J., Montgomery, M. K., Rajman, L., White, J. P., Teodoro, J. S., Wrann, C. D., Hubbard, B. P., et al. (2013) · Cell · doi:10.1016/j.cell.2013.11.037 · Cited by 1,482
Mitochondria-Targeted Small Molecule Therapeutics and Probes
Smith, R. A. J., Hartley, R. C., & Murphy, M. P. (2011) · Antioxidants & Redox Signaling · doi:10.1089/ars.2011.3969 · Cited by 399
Minimizing the damage: repair pathways keep mitochondrial DNA intact
Kazak, L., Reyes, A., & Holt, I. J. (2012) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm3439 · Cited by 388
Cisplatin induced Mitochondrial DNA damage in dorsal root ganglion neurons
Podratz, J. L., Knight, A. M., Ta, L. E., Staff, N. P., Gass, J. M., Genelin, K., Schlattau, A., Lathroum, L., & Windebank, A. J. (2011) · Neurobiology of Disease · doi:10.1016/j.nbd.2010.11.017 · Cited by 320
Lee, H.-Y., Choi, C. S., Birkenfeld, A. L., Alves, T. C., Jornayvaz, F. R., Jurczak, M. J., Zhang, D., Woo, D. K., Shadel, G. S., Ladiges, W., et al. (2010) · Cell Metabolism · doi:10.1016/j.cmet.2010.11.004 · Cited by 319
“Mitochondrial DNA Damage and Oxidative Protein Carbonylation Total genomic DNA was isolated from snap-frozen gastrocnemius skeletal muscle, and mtDNA copy number and integrity were determined using the QPCR method established by Van Houten and colleagues (Santos et al., 2006).”
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Metadata
- Authors
- Santos, J. H., Meyer, J. N., Mandavilli, B. S., & Houten, B.
- Venue
- Methods in Molecular Biology
- Publisher
- Humana Press
- Published
- 2008-09-19
- Volume
- 314
- Pages
- 183-199
- ISSN
- 1064-3745 (ISSN-L)
- OpenAlex
- W4235271617
- Cited by
- 285
- References
- 0
- FWCI
- 28.47
- Type
- book chapter
- Language
- en
- Open access
- closed
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- The paper
- Santos, J. H., Meyer, J. N., Mandavilli, B. S., & Houten, B. (2008). Quantitative PCR-Based Measurement of Nuclear and Mitochondrial DNA Damage and Repair in Mammalian Cells. Methods in Molecular Biology. https://doi.org/10.1385/1-59259-973-7%3A183
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