A New Method of the Visualization of the Double-Stranded Mitochondrial and Nuclear DNA

PLoS One. 2013 Jun 18;8(6):e66864. doi: 10.1371/journal.pone.0066864. Print 2013.

Abstract

The study describes the method of a sensitive detection of double-stranded DNA molecules in situ. It is based on the oxidative attack on the deoxyribose moiety by copper(I) in the presence of oxygen. We have shown previously that the oxidative attack leads to the formation of frequent gaps in DNA. Here we have demonstrated that the gaps can be utilized as the origins for an efficient synthesis of complementary labeled strands by DNA polymerase I and that such enzymatic detection of the double-stranded DNA is a sensitive approach enabling in-situ detection of both the nuclear and mitochondrial genomes in formaldehyde-fixed human cells.

MeSH terms

  • Cell Nucleus / genetics*
  • DNA / genetics*
  • DNA Polymerase I / metabolism
  • DNA, Mitochondrial / genetics*
  • HeLa Cells
  • Humans

Substances

  • DNA, Mitochondrial
  • DNA
  • DNA Polymerase I

Grants and funding

This work has been supported by the Technology Agency of the Czech Republic (TA03010598 and TA03010719). The infrastructural part of the project (Institute of Molecular and Translational Medicine) has been supported from the Operational Program Research and Development for Innovations (project CZ.1.05/2.1.00/01.0030). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.