Abstract
The alkaline comet assay (single cell gel electrophoresis) is the most widely used method for measuring DNA damage in eukaryotic cells (Neri et al., 2015). It detects strand breaks (SBs) and alkali-labile sites at frequencies from a few hundred to several thousand breaks per cell—a biologically useful range, extending from low endogenous damage levels to the extent of damage that can be inflicted experimentally without killing cells. Digestion of the nucleoids, after lysis, with certain lesion-specific repair endonucleases allows measurement of damage other than SBs; notably, formamidopyrimidine DNA glycosylase (FPG) has been widely used to detect altered purines, which are converted to breaks by the enzyme. Recently, (Cortes-Gutierrez et al., 2014) developed a two-dimensional Two-Tailed comet assay (TT-comet) that can differentiate between single-stranded (SSBs) and double-stranded DNA breaks (DSBs) in the same comets in sperm.
Highlights
Specialty section: This article was submitted to Genomic Assay Technology, a section of the journal Frontiers in Genetics
During the last 15 years, the comet assay has been extensively used in Drosophila melanogaster to test the genotoxicity of chemicals (Gaivão and Sierra, 2014)
This approach is very useful since Drosophila melanogaster is a valuable model for all kinds of processes related to human health, including DNA damage responses
Summary
Specialty section: This article was submitted to Genomic Assay Technology, a section of the journal Frontiers in Genetics. The alkaline comet assay (single cell gel electrophoresis) is the most widely used method for measuring DNA damage in eukaryotic cells (Neri et al, 2015).
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