彗星實驗
彗星實驗（英語：comet assay），又稱作單細胞凝膠電泳實驗或彗星電泳法（英語：Single Cell Gel Electrophoresis assay，SCGE）是一種對單個細胞檢測DNA損傷的實驗技術。彗星實驗最初由Östling和Johansson在1984年發明，此後由 Singh等在1988年改進，此後成為了對細胞內DNA損傷與修復、遺傳毒性生物監測的最常用的標準實驗技術。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]
參考文獻 編輯
- ^ 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).
延伸閱讀 編輯
- Dhawan & Anderson (2009): The Comet Assay in Toxicology. doi:10.1039/9781847559746
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- 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 （頁面存檔備份，存於網際網路檔案館）
- 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