Division for Biochemistry, School of Physical and Chemical Sciences, North-West University, Potchefstroom 2520, South Africa.
Anal Biochem. 2010 May 15;400(2):190-4. doi: 10.1016/j.ab.2010.02.008. Epub 2010 Feb 13.
The comet assay (single cell gel electrophoresis) is a cost-effective, sensitive, and simple technique that is traditionally used for analyzing and quantifying DNA damage in individual cells. The aim of this study was to determine whether the comet assay could be modified to detect changes in the levels of DNA methylation in single cells. We used the difference in methylation sensitivity of the isoschizomeric restriction endonucleases HpaII and MspI to demonstrate the feasibility of the comet assay to measure the global DNA methylation level of individual cells. The results were verified with the well-established cytosine extension assay. We were able to show variations in DNA methylation after treatment of cultured cells with 5-azacytidine and succinylacetone, an accumulating metabolite in human tyrosinemia type I.
彗星试验（单细胞凝胶电泳）是一种经济有效、敏感且简单的技术，传统上用于分析和量化单个细胞中的 DNA 损伤。本研究旨在确定彗星试验是否可以修改以检测单个细胞中 DNA 甲基化水平的变化。我们利用同裂酶内切核酸酶 HpaII 和 MspI 的甲基化敏感性差异，证明了彗星试验检测单个细胞整体 DNA 甲基化水平的可行性。结果用成熟的胞嘧啶延伸试验进行了验证。我们能够显示出在 5-氮杂胞苷和琥珀酰丙酮处理培养细胞后，人酪氨酸血症 I 中积累的代谢物，DNA 甲基化的变化。
