Analysis of DNA Double-Stranded Breaks Using the Comet Assay in Planarians

Paul G Barghouth; Salvador Rojas; Lacey R O'Dell; Andrew M Betancourt; Néstor J Oviedo

doi:10.1007/978-1-0716-2172-1_25

Analysis of DNA Double-Stranded Breaks Using the Comet Assay in Planarians

Methods Mol Biol. 2022:2450:479-491. doi: 10.1007/978-1-0716-2172-1_25.

Authors

Paul G Barghouth^{1

2}, Salvador Rojas^{1

2}, Lacey R O'Dell¹, Andrew M Betancourt¹, Néstor J Oviedo^{3

4

5}

Affiliations

¹ Department of Molecular & Cell Biology, University of California, Merced, CA, USA.
² Quantitative and Systems Biology Graduate Program, University of California, Merced, CA, USA.
³ Department of Molecular & Cell Biology, University of California, Merced, CA, USA. noviedo2@ucmerced.edu.
⁴ Quantitative and Systems Biology Graduate Program, University of California, Merced, CA, USA. noviedo2@ucmerced.edu.
⁵ Health Sciences Research Institute, University of California, Merced, CA, USA. noviedo2@ucmerced.edu.

Abstract

Comet assay provides the opportunity to detect and characterize DNA strand breaks. Cellular lysing followed by embedding in agarose slide is used to visualize under an electrical current migration patterns corresponding to DNA fragments of different sizes. Here we describe the process of detecting and characterizing DNA damage by Comet assay on planarians, which is a model organism commonly used to understand the process of whole-body regeneration, stem cell regulation, and adult tissue maintenance.

Keywords: Alkaline Comet Assay; DNA damage; Double-stranded breaks; Planarian.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Comet Assay
DNA / analysis
DNA Breaks, Double-Stranded
DNA Damage
Planarians* / genetics

Substances

DNA

Grants and funding

R01 GM132753/GM/NIGMS NIH HHS/United States