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

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity. / Vodenkova, Sona; Azqueta, Amaya; Collins, Andrew; Dusinska, Maria; Gaivao, Isabel; Møller, Peter; Opattova, Alena; Vodicka, Pavel; Godschalk, Roger W. L.; Langie, Sabine A. S.

In: Nature Protocols, Vol. 15, 2020, p. 3844–3878.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Vodenkova, S, Azqueta, A, Collins, A, Dusinska, M, Gaivao, I, Møller, P, Opattova, A, Vodicka, P, Godschalk, RWL & Langie, SAS 2020, 'An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity', Nature Protocols, vol. 15, pp. 3844–3878. https://doi.org/10.1038/s41596-020-0401-x

APA

Vodenkova, S., Azqueta, A., Collins, A., Dusinska, M., Gaivao, I., Møller, P., Opattova, A., Vodicka, P., Godschalk, R. W. L., & Langie, S. A. S. (2020). An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity. Nature Protocols, 15, 3844–3878. https://doi.org/10.1038/s41596-020-0401-x

Vancouver

Vodenkova S, Azqueta A, Collins A, Dusinska M, Gaivao I, Møller P et al. An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity. Nature Protocols. 2020;15:3844–3878. https://doi.org/10.1038/s41596-020-0401-x

Author

Vodenkova, Sona ; Azqueta, Amaya ; Collins, Andrew ; Dusinska, Maria ; Gaivao, Isabel ; Møller, Peter ; Opattova, Alena ; Vodicka, Pavel ; Godschalk, Roger W. L. ; Langie, Sabine A. S. / An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity. In: Nature Protocols. 2020 ; Vol. 15. pp. 3844–3878.

Bibtex

@article{3a0f42c987664fd98ea634b4b51a05d7,
title = "An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity",
abstract = "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.This protocol describes a comet-based in vitro assay for detecting base and nucleotide excision repair activity for use in cell culture model systems, human biomonitoring and clinical investigations, and animal studies, using isolated blood cells and various solid tissues.",
keywords = "OXIDATIVELY DAMAGED DNA, CANCER PATIENTS, POLYPHENOLIC COMPOUNDS, OCCUPATIONAL-EXPOSURE, INCISION ACTIVITY, EXPRESSION LEVELS, TELOMERE LENGTH, MINERAL FIBERS, CROSS-LINKS, CELLS",
author = "Sona Vodenkova and Amaya Azqueta and Andrew Collins and Maria Dusinska and Isabel Gaivao and Peter M{\o}ller and Alena Opattova and Pavel Vodicka and Godschalk, {Roger W. L.} and Langie, {Sabine A. S.}",
year = "2020",
doi = "10.1038/s41596-020-0401-x",
language = "English",
volume = "15",
pages = "3844–3878",
journal = "Nature Protocols (Print)",
issn = "1754-2189",
publisher = "nature publishing group",

}

RIS

TY - JOUR

T1 - An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity

AU - Vodenkova, Sona

AU - Azqueta, Amaya

AU - Collins, Andrew

AU - Dusinska, Maria

AU - Gaivao, Isabel

AU - Møller, Peter

AU - Opattova, Alena

AU - Vodicka, Pavel

AU - Godschalk, Roger W. L.

AU - Langie, Sabine A. S.

PY - 2020

Y1 - 2020

N2 - 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.This protocol describes a comet-based in vitro assay for detecting base and nucleotide excision repair activity for use in cell culture model systems, human biomonitoring and clinical investigations, and animal studies, using isolated blood cells and various solid tissues.

AB - 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.This protocol describes a comet-based in vitro assay for detecting base and nucleotide excision repair activity for use in cell culture model systems, human biomonitoring and clinical investigations, and animal studies, using isolated blood cells and various solid tissues.

KW - OXIDATIVELY DAMAGED DNA

KW - CANCER PATIENTS

KW - POLYPHENOLIC COMPOUNDS

KW - OCCUPATIONAL-EXPOSURE

KW - INCISION ACTIVITY

KW - EXPRESSION LEVELS

KW - TELOMERE LENGTH

KW - MINERAL FIBERS

KW - CROSS-LINKS

KW - CELLS

U2 - 10.1038/s41596-020-0401-x

DO - 10.1038/s41596-020-0401-x

M3 - Journal article

C2 - 33199871

VL - 15

SP - 3844

EP - 3878

JO - Nature Protocols (Print)

JF - Nature Protocols (Print)

SN - 1754-2189

ER -

ID: 252467085