The International Comet Assay Workshop

Abstract

The Comet (single cell gel electrophoresis) assay primarily measures DNA strand breakage in single cells. Briefly, cells are suspended in low melting point agarose on a microscope slide. The slides are put in lysing buffer to allow the DNA to unwind and then in electrophoresis buffer. During electrophoresis the broken DNA moves towards the anode forming a Comet tail, with the greater the extent of damage, the greater the tail. Assays can be conducted under neutral or alkaline (> pH 13) conditions. Double-strand breaks are measured under neutral conditions and single-strand breaks under alkaline conditions, where abasic sites and other alkali-labile sites or intermediates in base or nucleotide excision repair can also be detected. There are several good reviews concerning the assay.