Abstract
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.
Full Text
The Full Text of this article is available as a PDF (57.8 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ager D. D., Dewey W. C., Gardiner K., Harvey W., Johnson R. T., Waldren C. A. Measurement of radiation-induced DNA double-strand breaks by pulsed-field gel electrophoresis. Radiat Res. 1990 May;122(2):181–187. [PubMed] [Google Scholar]
- Blöcher D., Einspenner M., Zajackowski J. CHEF electrophoresis, a sensitive technique for the determination of DNA double-strand breaks. Int J Radiat Biol. 1989 Oct;56(4):437–448. doi: 10.1080/09553008914551591. [DOI] [PubMed] [Google Scholar]
- Rosemann M., Kanon B., Konings A. W., Kampinga H. H. An image analysis technique for detection of radiation-induced DNA fragmentation after CHEF electrophoresis. Int J Radiat Biol. 1993 Aug;64(2):245–249. doi: 10.1080/09553009314551361. [DOI] [PubMed] [Google Scholar]
