Single cell gel electrophoresis, also known as the comet assay, is widely used for the detection and measurement of DNA strand breaks. With the addition of a step in which DNA is incubated with specific endonucleases recognising damaged bases, these lesions can be measured, too. In the standard protocol, electrophoresis is carried out at high pH. If, instead, electrophoresis is in neutral buffer, the effect of DNA damage seems to be much reduced--either because alkaline conditions are needed to reveal certain lesions, or because the effect of the same number of breaks on DNA migration is greater at high pH. A lower sensitivity can be useful in some circumstances, as it extends the range of DNA damage levels over which the assay can be used. Here we compare the performance of standard and modified techniques with a variety of DNA-damaging agents and offer possible explanations for the differences in behaviour of DNA under alternative electrophoretic conditions.

Institute of Experimental Botany of Czech Academy of Sciences Praha Czech Republic

Citace poskytuje Crossref.org

RAD51 and RAD51B Play Diverse Roles in the Repair of DNA Double Strand Breaks in Physcomitrium patens

DNA damage triggers reprogramming of differentiated cells into stem cells in Physcomitrella

Evolutionarily Distant Streptophyta Respond Differently to Genotoxic Stress

Bridging Plant and Human Radiation Response and DNA Repair through an In Silico Approach

Genotoxin induced mutagenesis in the model plant Physcomitrella patens