Pulsed field gel electrophoresis (PFGE) is widely used to measure DNA double strand breaks (dsb). The DNA of cultured cells can be prelabelled with radioactivity, which helps greatly in detection and quantitation of DNA dsb. However, this approach cannot be used with non-cycling cells from biopsy material. We describe a method which uses SYBR Green I to stain DNA in dried agarose gels. DNA is detected and analysed using readily available camera equipment and image analysis software. This method is as sensitive as [3H]thymidine prelabelling of cells and allows DNA dsb to be measured simply and economically in non-cycling cells.
Bibliographical noteFunding Information:
A.E.K. is supported on a grant (Principle Investigator J.H.Hendry) from the Christie Hospital NHS Trust Endowment Fund. We wish to thank Professor Jolyon Hendry, Dr Geoff Margison and Mr Tim Ward for their help throughout this work.