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An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity

S. Vodenkova, A. Azqueta, A. Collins, M. Dusinska, I. Gaivão, P. Møller, A. Opattova, P. Vodicka, RWL. Godschalk, SAS. Langie

. 2020 ; 15 (12) : 3844-3878. [pub] 20201116

Jazyk angličtina Země Velká Británie

Typ dokumentu časopisecké články, práce podpořená grantem, Research Support, U.S. Gov't, Non-P.H.S.

Perzistentní odkaz   https://www.medvik.cz/link/bmc21011806
E-zdroje Online Plný text

NLK ProQuest Central od 2006-06-01 do Před 1 rokem
Health & Medicine (ProQuest) od 2006-06-01 do Před 1 rokem

Odkazy

PubMed 33199871
DOI 10.1038/s41596-020-0401-x
Knihovny.cz E-zdroje

This optimized protocol (including links to instruction videos) describes a comet-based in vitro DNA repair assay that is relatively simple, versatile, and inexpensive, enabling the detection of base and nucleotide excision repair activity. Protein extracts from samples are incubated with agarose-embedded substrate nucleoids ('naked' supercoiled DNA) containing specifically induced DNA lesions (e.g., resulting from oxidation, UVC radiation or benzo[a]pyrene-diol epoxide treatment). DNA incisions produced during the incubation reaction are quantified as strand breaks after electrophoresis, reflecting the extract's incision activity. The method has been applied in cell culture model systems, human biomonitoring and clinical investigations, and animal studies, using isolated blood cells and various solid tissues. Once extracts and substrates are prepared, the assay can be completed within 2 d.

Citace poskytuje Crossref.org

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