SYBR Green I staining of pulsed field agarose gels is a sensitive and inexpensive way of quantitating DNA double-strand breaks in mammalian cells.
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.
Kiltie
A E
AE
Department of Experimental Radiation Oncology, Paterson Institute for Cancer Research, Manchester M20 4BX, UK.
Ryan
A J
AJ
D000818:Animals; D010582:Bacteriophage lambda / Q000235:genetics; D052160:Benzothiazoles; D004247:DNA / Q000032:analysis; D004249:DNA Damage; D004279:DNA, Viral / Q000032:analysis*; D003959:Diamines; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D005456:Fluorescent Dyes; D008322:Mammals; D009930:Organic Chemicals; D011804:Quinolines; D012680:Sensitivity and Specificity; D012685:Sepharose; D013194:Staining and Labeling
D052160:Benzothiazoles; D004279:DNA, Viral; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D004247:DNA; D012685:Sepharose