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Tracking Mitochondrial DNA In Situ

  • Anna Ligasová
  • Karel KobernaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1351)

Abstract

The methods of the detection of (1) non-labeled and (2) BrdU-labeled mitochondrial DNA (mtDNA) are described. They are based on the production of singlet oxygen by monovalent copper ions and the subsequent induction of DNA gaps. The ends of interrupted DNA serve as origins for the labeling of mtDNA by DNA polymerase I or they are utilized by exonuclease that degrades DNA strands, unmasking BrdU in BrdU-labeled DNA. Both methods are sensitive approaches without the need of additional enhancement of the signal or the use of highly sensitive optical systems.

Key words

Mitochondrial DNA Mitochondrial DNA replication Copper ions Biotin-16-dUTP Singlet oxygen 

Notes

Acknowledgement

This work was supported by the Technology Agency of the Czech Republic (TA03010598, TA03010719, and TE02000058), the Ministry of Health of the Czech Republic [AZV 15-31604A] and the Ministry of Education, Youth and Sports (LO1304).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Faculty of Medicine, Institute of Molecular and Translational MedicinePalacký UniversityOlomoucCzech Republic

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