title: Tracking Mitochondrial DNA In Situ.
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.
authors.affiliation: Faculty of Medicine, Institute of Molecular and Translational Medicine, Palacký University, Hněvotínska 5, Olomouc, 77900, Czech Republic.
authors.forename: Anna
authors.initials: A
authors.lastname: Ligasová
authors.affiliation: Faculty of Medicine, Institute of Molecular and Translational Medicine, Palacký University, Hněvotínska 5, Olomouc, 77900, Czech Republic. karel.koberna@upol.cz.
authors.forename: Karel
authors.initials: K
authors.lastname: Koberna
mesh_terms: D000818:Animals; D001710:Biotin / Q000737:chemistry; D001973:Bromodeoxyuridine / Q000737:chemistry; D002478:Cells, Cultured; D003300:Copper / Q000737:chemistry; D004256:DNA Polymerase I / Q000378:metabolism; D004261:DNA Replication; D004272:DNA, Mitochondrial / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006801:Humans; D007211:Indoles / Q000737:chemistry; D008928:Mitochondria / Q000235:genetics*; D010100:Oxygen / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D004272:DNA, Mitochondrial; D007211:Indoles; D049452:Green Fluorescent Proteins; C007293:DAPI; D001710:Biotin; D003300:Copper; D004256:DNA Polymerase I; D001973:Bromodeoxyuridine; D010100:Oxygen
keywords: Biotin-16-dUTP; Copper ions; Mitochondrial DNA; Mitochondrial DNA replication; Singlet oxygen