The International Comet Assay Workshop.
The Comet (single cell gel electrophoresis) assay primarily measures DNA strand breakage in single cells (Singh et al., 1988). 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 (McKelvey-Martin et al., 1993; Fairbairn et al., 1995; Tice, 1995).
Anderson
D
D
BIBRA International, Carshalton, Surrey, UK. danderson@bibra.co.uk
Plewa
M J
MJ
D002457:Cell Extracts / Q000737:chemistry* / Q000235:genetics*; D002460:Cell Line; D002985:Clinical Protocols; D004249:DNA Damage; D004260:DNA Repair; D004587:Electrophoresis, Agar Gel / Q000379:methods* / Q000639:trends; D005854:Germ Cells / Q000737:chemistry; D006801:Humans; D006822:Hybrid Cells / Q000737:chemistry
D002457:Cell Extracts