Optimization of single-cell gel electrophoresis (SCGE) method for study of liver single cell DNA in Caspian kutum (Rutilus frisii kutum)

Document Type : Research Paper



Single-cell gel electrophoresis (SCGE) is a method that used to evaluate DNA damage in single cells. This method due to its great precision and high operating speed, considered than to other methods. The present study was performed to optimize some of the alkaline comet assay steps by using Caspian kutum )Rutilus frisii kutum( liver single cells. Hence, one batch of the fish fingerlings was used for the experiment (n=10, 7±1g) and fish liver tissues were dissected and transferred to freezer (-80C°) until starting the optimizing procedure and other common methods for comet assay tests. In optimization of conventional method for obtain single cells, centrifuging steps were deleted. Also, the solidification of gel on slides decreased and lysing time of cell membrane increased. Finally, a simple way to archive slides stained with ethidium bromide was present for at least two weeks. In addition to reducing experimental time and consumption materials, the optimized method led to lower cell damage and possibility of artificial error. No significant differences were observed between comet indexes, such as comet length (px), comet height (px), tail length (px), DNA% in tail and tail moment, achieved from the optimized and conventional method.