Measuring the repair of H2O2-induced DNA single strand breaks using the single cell gel assay.
The single cell gel (SCG) assay is for the detection of DNA single strand breaks and alkali labile sites in single cells. This investigation showed how the technique had an effective application in examining DNA single strand break repair. DNA from cultured human cells was damaged by treating the cells with H2O2, and samples were allowed to repair for various time intervals. The cells were then electrophoresed in a gel on a microscope slide, and the sizes of the individual DNA 'comets' were measured using laser scanning microscopy. Analysis of the comets indicated that actual DNA repair could be monitored over time. The SCG assay will help the study of DNA damage and repair, which will aid our understanding of the complex processes of mutagenesis and carcinogenesis.
Meyers
C D
CD
Department of Microbiology, Brigham Young University, Provo, Utah 84604.
Fairbairn
D W
DW
O'Neill
K L
KL
D002460:Cell Line; D004249:DNA Damage; D004260:DNA Repair / Q000502:physiology*; D004277:DNA, Single-Stranded / Q000187:drug effects* / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007091:Image Processing, Computer-Assisted; D008853:Microscopy / Q000379:methods; D012984:Software; D013997:Time Factors
D004277:DNA, Single-Stranded; D006861:Hydrogen Peroxide