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插层分子在银离子存在下检测甲基化 DNA 的应用。

Application of intercalating molecules in detection of methylated DNA in the presence of silver ions.

机构信息

Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, Iran.

出版信息

Methods Appl Fluoresc. 2019 May 16;7(3):035005. doi: 10.1088/2050-6120/ab025b.

Abstract

The extent of DNA structural perturbation by silver ions is different in methylated and non-methylated DNA. Here, we explored the interaction of eight convenient DNA interacting molecules with methylated and non-methylated short GC rich oligonucleotides in the presence and absence of silver ions. Acridine orange, DAPI, Doxorubicin, Ethidium bromide, Hoechst 33342, Methylene blue, PicoGreen, and Propidium iodide are tested for their ability to distinguish methylated and non-methylated DNA. Among them, Ethidium bromide, Methylene blue, and PicoGreen were able to discriminate between methylated and non-methylated DNA, while DAPI and Hoechst 33342 were only able to discriminate with the aid of silver ions. A detection method is proposed using Ethidium bromide in which the silver-treated sample of DNA exposed different fluorescence intensity from the untreated one on the base of its methylation state. This phenomenon was sequence-dependent and could provide a sensing platform with a detection limit of about 4fi0 nM.

摘要

银离子对 DNA 结构的干扰程度在甲基化和非甲基化 DNA 中有所不同。在这里,我们研究了在存在和不存在银离子的情况下,八种方便的与 DNA 相互作用的分子与甲基化和非甲基化的富含 GC 的短寡核苷酸的相互作用。吖啶橙、DAPI、阿霉素、溴化乙锭、Hoechst 33342、亚甲蓝、PicoGreen 和碘化丙啶被测试其区分甲基化和非甲基化 DNA 的能力。其中,溴化乙锭、亚甲蓝和 PicoGreen 能够区分甲基化和非甲基化 DNA,而 DAPI 和 Hoechst 33342 只有在银离子的帮助下才能区分。提出了一种使用溴化乙锭的检测方法,其中基于 DNA 的甲基化状态,经银处理的 DNA 样品与未经处理的样品在荧光强度上表现出不同。这种现象是序列依赖性的,可以提供一个检测限约为 4fi0 nM 的传感平台。

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