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

Anna Ligasová, Dmytro Strunin, Karel Koberna

Journal: PloS one 2017;8(6):e66864

PMID: 23825578

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.

Address: Laboratory of Experimental Medicine, Institute of Molecular and Translational Medicine, Faculty of Medicine, Palacký University in Olomouc, Olomouc, Czech Republic.
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