title: Atomic scissors: a new method of tracking the 5-bromo-2'-deoxyuridine-labeled DNA in situ.
abstract: A new method of the light microscopy detection of BrdU-labeled DNA in situ is described. It is based on the oxidative attack at the deoxyribose moiety by copper(I) in the presence of oxygen, which leads to the abstraction of hydrogen atom from deoxyribose culminating in the elimination of the nucleobase, scission of the nucleic-acid strand and formation of frequent gaps. The gaps allow the reaction of the antibodies with the commonly used markers of replication (e.g. 5-bromo-2'-deoxyuridine), which are otherwise masked. The method developed makes it possible to detect nuclear and mitochondrial DNA replication efficiently. In most cases, it does not inhibit effective protein detections and in addition enables simultaneous localization of newly-synthesized RNA. The alternative presently-used methods result in protein denaturation and/or extensive DNA cleavage followed by the DNA-bound proteins peeling off.
authors.affiliation: Department of Molecular Cytology and Cytometry, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic. ligasova@gmail.com
authors.forename: Anna
authors.initials: A
authors.lastname: Ligasová
authors.forename: Dmytro
authors.initials: D
authors.lastname: Strunin
authors.forename: Radek
authors.initials: R
authors.lastname: Liboska
authors.forename: Ivan
authors.initials: I
authors.lastname: Rosenberg
authors.forename: Karel
authors.initials: K
authors.lastname: Koberna
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000737:chemistry; D001973:Bromodeoxyuridine / Q000737:chemistry* / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D019327:Copper Sulfate / Q000737:chemistry; D053837:DNA Cleavage; D004261:DNA Replication; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics; D003850:Deoxyribonuclease I; D019084:Fluorescent Antibody Technique, Indirect; D006367:HeLa Cells; D006801:Humans; D010084:Oxidation-Reduction; D010100:Oxygen / Q000737:chemistry; D013194:Staining and Labeling; D013482:Superoxide Dismutase / Q000737:chemistry; D013481:Superoxides / Q000737:chemistry
chemical_list: D004272:DNA, Mitochondrial; D013481:Superoxides; D013482:Superoxide Dismutase; D003850:Deoxyribonuclease I; D001973:Bromodeoxyuridine; D019327:Copper Sulfate; D001205:Ascorbic Acid; D010100:Oxygen