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. 2025:2954:273-283.
doi: 10.1007/978-1-0716-4698-4_16.

Methods to Evaluate DNA Damage in Spermatozoa

Jacinta H Martin  1   2 , Natalie A Trigg  3   4   5   6 , Leah Calvert  3   4 , Amanda L Anderson  3   4 , Tessa Lord  3   4 , John E Schjenken  3   4 , Shaun D Roman  7 , Geoffry N De Iuliis  3   4 , Brett Nixon  3   4
Affiliations

Methods to Evaluate DNA Damage in Spermatozoa

Jacinta H Martin et al. Methods Mol Biol. 2025.

Abstract

Studying DNA damage in spermatozoa allows researchers fundamental insight into the molecular mechanisms underlying the cellular response of sperm to stressors and provides an opportunity for understanding novel mechanisms for mitigating this type of damage. Furthermore, since compromised genetic integrity is recognized as a leading contributor to infertility phenotypes, reliable methods for assessing DNA damage in populations of spermatozoa are essential in research and clinical laboratories alike. Here, we describe two methods, the sperm chromatin structure assay (SCSA) and alkaline comet assay, designed to assess both single- and double-stranded DNA damage within the spermatozoon.

Keywords: Acridine orange (AO); Alkaline comet assay; DNA; Double-strand breaks (DSB); Flow cytometry; Genetic integrity; Human; Mouse/mice; Single-strand breaks (SSB); Sperm chromatin structure assay (SCSA).

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