彗星实验

彗星实验（英语：comet assay），又称作单细胞凝胶电泳实验或彗星电泳法（英语：Single Cell Gel Electrophoresis assay，SCGE）是一种对单个细胞检测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，此后成为了对细胞内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。步骤包括将细胞包覆在低融点的琼脂糖中, 在中性或碱性下(pH > 13)溶解细胞, 最后以凝胶电泳将包覆的已溶解细胞分离解析. 之所以称为彗星, 是指细胞DNA在凝胶电泳解析分离后呈现的样子有点像彗星.^[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
Avishai, Nanthawan; Rabinowitz, Claudette; Moiseeva, Elisabeth & Rinkevich, Baruch (2002): Genotoxicity of the Kishon River, Israel: the application of an in vitro cellular assay. Mutation Research 518(1): 21–37. doi:10.1016/S1383-5718(02)00069-4 (HTML abstract)
Collins, A.R.; Dobson, V.L.; Dusinska, M.; Kennedy, G. & Stetina, R. (1997): The comet assay: what can it really tell us? Mutation Research 375(2): 183-193. doi:10.1016/S0027-5107(97)00013-4 (HTML abstract)
Klaude, M.; Eriksson, S.; Nygren, J. & Ahnstrom, G. (1996): The comet assay: mechanisms and technical considerations. Mutation Research 363(2): 89-96. doi:10.1016/0921-8777(95)00063-1
McKelvey-Martin, Valerie J.; Ho, Edwin T.; McKeown, Stephanie R.; Johnston, S. Robin; McCarthy, Patsy J.; Rajab, Nor Fasilah & Downes, C. Stephen (1993): Emerging applications of the single cell gel electrophoresis (Comet) assay. I. Management of invasive transitional cell human bladder carcinoma. II. Fluorescent in situ hybridization Comets for the identification of damaged and repaired DNA sequences in individual cells. Mutagenesis 13(1): 1-8. PDF fulltext
Olive, P.L.; Wlodek, D. & Banath, J.P. (1991): DNA double-strand breaks measured in individual cells subjected to gel electrophoresis. Cancer Research 51(17): 4671-4676. PDF fulltext （页面存档备份，存于互联网档案馆）
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

[1] 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.

[2] 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).

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