Atomic Scissors: a New Method of Tracking the 5-bromo-2'-deoxyuridine-labeled DNA in Situ

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2013 Jan 10

PMID 23300711

Citations 16

Authors

Anna Ligasova

Dmytro Strunin

Radek Liboska

Ivan Rosenberg

Karel Koberna

Affiliations

Soon will be listed here.

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.

Citing Articles

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A basal-level activity of ATR links replication fork surveillance and stress response.

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