| 近红外荧光电泳迁移率变动分析的实验方法 |
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| 引用本文： | 高婧,刘颖勋,王进科.近红外荧光电泳迁移率变动分析的实验方法[J].生物技术通讯,2011,22(1):71-76. |
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| 作者姓名： | 高婧 刘颖勋 王进科 |
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| 作者单位： | 东南大学,生物电子学国家重点实验室,生物科学与医学工程学院生物技术与材料实验中心,江苏,南京,210096 |
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| 基金项目： | 国家自然科学基金(60871014) |
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| 摘 要： | 目的:分析最新近红外荧光标记(NIRF)电泳迁移率变动分析(EMSA)技术的优缺点,在此基础上对NIRF-EMSA技术进行实验方法的改进。方法:设计了3种NIRF-EMSA实验方法,即扫胶法、染胶法和扫膜法,通过实验比较了3种方法的实验程序复杂性、优缺点及检测成本,并将3种方法与化学发光EMSA进行了比较。结果:采用3种方法都可以取得良好的EMSA检测效果,但比较而言,间接扫膜法不仅效果良好,而且实验成本最低。结论:间接扫膜法是值得推广应用的最经济便捷的基于Odyssey红外荧光成像系统的近红外荧光EMSA实验方法。
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| 关 键 词： | 近红外荧光电泳迁移率变动分析 Odyssey红外荧光成像系统 间接扫膜法 |
Experimental Methods of Near-Infrared Fluorescence Electrophoresis Mobility Shift Assay |
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| GAO Jing,LIU Ying-Xun,WANG Jin-Ke.Experimental Methods of Near-Infrared Fluorescence Electrophoresis Mobility Shift Assay[J].Letters in Biotechnology,2011,22(1):71-76. |
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| Authors: | GAO Jing LIU Ying-Xun WANG Jin-Ke |
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| Institution: | GAO Jing,LIU Ying-Xun,WANG Jin-Ke State Key Laboratory of Bioelectronics,Experimental Center of Biotechnology and Biomaterials,School of Biological Sciences and Medical Engineering,Southeast University,Nanjing 210096,China |
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| Abstract: | Objective:The electrophoresis mobility shift assay(EMSA) is golden standard technique for studying the interaction of DNA and protein.The merits and limitations of the latest near-infrared fluorescence-labeled EMSA(NIRF-EMSA) were briefly analyzed.On this base,experiments were designed to improve the NIRF-EMSA protocol for routine application.Methods:We tried three NIRF-EMSA protocols,including scanning gel,staining gel and scanning membrane,and systematically evaluated the complexity of experimental proced... |
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| Keywords: | near-infrared fluorescence electrophoresis mobility shift assay Odyssey infrared imaging system scanning membrane |
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