DNA Double-strand Break Analysis by CHEF Electrophoresis

Summary

Pulsed-field gel electrophoresis (PFGE) has become a powerful tool for the study of high-molecular-weight DNA. In this technical note the application of clamped homogeneous electrical field (CHEF) electrophoresis for the determination of DNA double-strand breaks is described, and a number of parameters that influence the sensitivity of the assay are discussed. They include the concentration of the cell sample, agarose concentration in the plug and the electrophoresis gel, lysis conditions, pulse time, electric field gradient and the temperature of electrophoresis.