Detection of DNA damage by alkaline comet assay in mouse colonic mucosa

Elisa Boutet-Robinet; Maria M Haykal; Saleha Hashim; Teresa Frisan; Océane C B Martin

doi:10.1016/j.xpro.2021.100872

Detection of DNA damage by alkaline comet assay in mouse colonic mucosa

STAR Protoc. 2021 Oct 23;2(4):100872. doi: 10.1016/j.xpro.2021.100872. eCollection 2021 Dec 17.

Authors

Elisa Boutet-Robinet¹, Maria M Haykal², Saleha Hashim¹, Teresa Frisan^{3

4}, Océane C B Martin⁵

Affiliations

¹ Toxalim (Research Centre in Food Toxicology), Université de Toulouse, INRAE, ENVT, INP-Purpan, UPS, Toulouse, France.
² INSERM U981, LabEx LERMIT, Université Paris 12 Sud, Gustave Roussy Research Center, Department of Molecular Medicine, Villejuif, France.
³ Department of Molecular Biology, Umeå University, Umeå, Sweden.
⁴ Umeå Centre for Microbial Research (UCMR), Umeå University, Umeå, Sweden.
⁵ University of Bordeaux, INSERM, BaRITOn, U1053, 33000 Bordeaux, France.

Abstract

We recently characterized the association between DNA damage and immunoresponse in vivo in colonic mucosa of mice infected with a Salmonella Typhimurium strain expressing a genotoxin, known as typhoid toxin. In this protocol, we describe the specific steps for assessing DNA damage by the alkaline comet assay of colonic mucosal samples. The description of the comet assay protocol follows the international guidelines (Minimum Information for Reporting on the Comet Assay [Moller et al., 2020]). For complete details on the use and execution of this protocol, please refer to Martin et al. (2021).

Keywords: Cell isolation; Cell-based Assays; Microscopy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques
Colon / cytology*
Comet Assay / methods*
DNA Damage / genetics*
Intestinal Mucosa / cytology*
Mice