مرکز منطقه ای اطلاع رساني علوم و فناوري - Alternative safe stains for single cell gel electrophoresis assay

Abstract :

Aim. Single cell gel electrophoresis or the other common name Comet assay technique is a sensitive, fast and cheap cytogenetic method for DNA damage assessment. Method consists of embedding cells to the agarose on microscope slides, electrophoresis following lysis and staining with fluorescent dyes for DNA damage visualization basically. At the staining stage intercalator dyes ethidium bromide and propidium iodide are used widely. However because of the mutagenic and possible carcinogenic properties of these dyes, they must be used very carefully. Method. In this study applicability of two safe non-carcinogenic dyes for DNA visualization in single cell gel electrophoresis is investigated. Lymphocytes from blood sample of a healthy volunteer donor were separated and propagated in cell culture, divided into two, one of them used as control and the other is irradiated 2Gy at a 0,527 kGy/h dose rated 137Cs source. Alkaline comet assay was applied and slides were stained with ethidium bromide and propidium iodide also with alternative SafeView and GelStar dyes. Results. Optained data shows that, when compared with the conventional ones these two alternative dyes worked efficiently. Conclusion. Results of this study showed that SafeView and GelStar dyes which are used in classical agarose gel electrophoresis method for DNA and RNA staining, can also be used efficiently in single cell gel electrophoresis for the first time in literature.