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O-Hydroxylamine-Coupled Alkaline Gel Electrophoresis Assay for the Detection and Measurement of DNA Single-Strand Breaks

  • April M. Luke
  • Jun NakamuraEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 920)

Abstract

The ability to detect and measure DNA single-strand breaks has been the aim of numerous assays developed to assess genotoxicity. These methods often rely on alkaline conditions to denature DNA. However, alkaline treatment of DNA also introduces artifactual SSBs through the cleavage of alkali-labile sites resulting in confounded data. Here, we describe a modified alkaline gel electrophoresis assay coupled with a neutral O-hydroxylamine to obtain the measurement of true SSB formation.

Key words

Agarose Alkaline Electrophoresis O-hydroxylamine Single-strand breaks 

Notes

Acknowledgements

This work was supported by National Institute of Environmental Health Science grants T32-ES07126, P42-ES05948, and P30-ES10126 and National Cancer Institute grant T32-CA72319.

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Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  1. 1.Department of Environmental Sciences and EngineeringUniversity of North CarolinaChapel HillUSA

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