彗星实验
彗星实验（英語：comet assay），又称作单细胞凝胶电泳实验或彗星电泳法（英語：Single Cell Gel Electrophoresis assay，SCGE）是一种對单个细胞检测DNA损伤的实验技术。彗星实验最初由Östling和Johansson在1984年发明，此后由 Singh等在1988年改进，此后成为了對細胞內DNA损伤与修复、遗传毒性生物监测的最常用的标准实验技术。步驟包括將細胞包覆在低融點的瓊脂糖中, 在中性或鹼性下(pH > 13)溶解細胞, 最後以凝膠電泳將包覆的已溶解細胞分離解析. 之所以稱為彗星, 是指細胞DNA在凝膠電泳解析分離後呈現的樣子有點像彗星.[1][2]
参考文献
- ^ Tice, R. R. Single Cell Gel/Comet Assay: Guidelines for in vitro and in vivo Genetic Toxicology Testing. Environmental and Molecular Mutagenesis. 2000, 35 (3): 206–21. PMID 10737956. doi:10.1002/(SICI)10982280.
- ^ Nandhakumar S, Parasuraman S, Shanmugam M M, Rao K R, Chand P, Bhat B V. Evaluation of DNA damage using single-cell gel electrophoresis (Comet Assay).
延伸阅读
- Dhawan & Anderson (2009): The Comet Assay in Toxicology. doi:10.1039/9781847559746
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- Rojas, E.; Lopez, M.C. & Valverde, M. (1999): Single cell gel electrophoresis: methodology and applications. Journal of Chromatography B 722(1-2): 225-254. doi:10.1016/S0378-4347(98)00313-2 (HTML abstract)
- Singh, N.P.; McCoy, M.T.; Tice, R.R. & Schneider, E.L. (1988): A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research 175(1): 184-191. doi:10.1016/0014-4827(88)90265-0
- Tice, R.R.; Agurell, E.; Anderson, D.; Burlinson, B.; Hartmann, A.; Kobayashi, H.; Miyamae, Y.; Rojas, E.; Ryu, J.-C. & Sasaki, Y.F. (2000): Single Cell Gel/Comet Assay: Guidelines for In Vitro and In Vivo Genetic Toxicology Testing. Environmental and Molecular Mutagenesis 35(3): 206-221. doi:10.1002/(SICI)10982280