Related Experiment Video
Updated: Oct 14, 2025
In Vivo Alkaline Comet Assay and Enzyme-modified Alkaline Comet Assay for Measuring DNA Strand Breaks and Oxidative DNA Damage in Rat Liver
Published on: May 4, 2016
Detection of DNA damage by alkaline comet assay in mouse colonic mucosa
Elisa Boutet-Robinet1, Maria M Haykal2, Saleha Hashim1
1Toxalim (Research Centre in Food Toxicology), Université de Toulouse, INRAE, ENVT, INP-Purpan, UPS, Toulouse, 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).
Insights
This study details a method to detect DNA damage in mouse colons after Salmonella infection. The alkaline comet assay protocol helps understand the link between genotoxins and immune responses.
Area of Science:
- Microbiology
- Immunology
- Genotoxicology
Background:
- Salmonella Typhimurium infection can cause DNA damage in the colon.
- Typhoid toxin, expressed by some Salmonella strains, is a genotoxin.
- Understanding DNA damage is crucial for studying the immune response to infection.
Purpose of the Study:
- To provide a detailed protocol for assessing DNA damage in mouse colonic mucosa.
- To facilitate research on the association between DNA damage and immunoresponse in vivo.
- To standardize the alkaline comet assay for genotoxicity studies.
Main Methods:
- Alkaline comet assay on colonic mucosal samples from infected mice.
- Utilizing a Salmonella Typhimurium strain expressing typhoid toxin.
- Following international guidelines for reporting the comet assay.
Main Results:
- The protocol enables reliable assessment of DNA damage.
- It allows for the characterization of DNA damage in the context of Salmonella infection.
- The method is described in detail for reproducibility.
Conclusions:
- The alkaline comet assay is a valuable tool for studying DNA damage in infectious disease models.
- This protocol aids in investigating the interplay between genotoxicity and host immune responses.
- Standardized methods are essential for advancing research in this field.
