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Quantification of DNA Damage by Real-Time qPCR

  • Wei Wang
  • Katja Scheffler
  • Ying Esbensen
  • Lars EideEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1351)

Abstract

This chapter describes the use of real-time qPCR to quantify damages in genomic DNA. The method is based on the ability of a lesion in one strand to inhibit restriction enzyme digestion of double-stranded DNA. Subsequent amplification of the complementary strand after restriction cleavage gives a quantitative measure of the damage content in that site (Real-time qPCR Analysis of Damage Frequency; RADF). We compare the RADF assay with the commonly used technique to assess damages by their ability to inhibit amplification of a large PCR fragment relative to a short PCR fragment. The RADF method described here is quick, accurate and allows the detection of nuclear and mitochondrial DNA damage in detailed regions.

Key words

DNA damage mtDNA DNA repair 

Notes

Acknowledgements

The work was supported by the Norwegian Research Council.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Wei Wang
    • 1
    • 2
  • Katja Scheffler
    • 1
    • 2
  • Ying Esbensen
    • 3
  • Lars Eide
    • 1
    Email author
  1. 1.Department of Medical Biochemistry, Institute of Clinical MedicineOslo University Hospital and University of OsloOsloNorway
  2. 2.Department of MicrobiologyOslo University Hospital and University of OsloOsloNorway
  3. 3.Institute of Clinical Epidemiology and Molecular BiologyAkershus University Hospital and University of OsloLørenskogNorway

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