The single cell gel electrophoresis or Comet assay is a sensitive, reliable, and rapid method for DNA double- and single-strand breaks, alkali-labile sites and delayed repair site detection, in eukaryotic individual cells. Given its overall characteristics, this method has been widely used over the past few years in several different areas. In this paper we review the studies published to date about the principles, the basic methodology with currently used variations. We also explore the applications of this assay in: genotoxicology, clinical area, DNA repair studies, environmental biomonitoring and human monitoring.