Supported lipid bilayers and DNA curtains for high-throughput single-molecule studies.
Supported lipid bilayers and DNA curtains for high-throughput single-molecule studies.
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DOI：
10.1007/978-1-61779-129-1_26
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Greene, Eric C
中科院分区：
文献类型：
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作者：
Finkelstein, Ilya J;Greene, Eric C
蛋白质- DNA相互作用的单分子研究继续产生关于许多DNA加工途径的新信息。例如，基于光学显微镜的技术允许实时观察与DNA底物相互作用的蛋白质，这反过来又允许直接了解反应机制。然而，这些实验在技术上仍然具有挑战性，并且由于缺乏稳定的发色团和难以获得统计上显著的观察结果而受到限制。在这个方案中，我们描述了一个新的，高通量的，纳米制造的实验平台，能够在小时的时间尺度上实时成像数百个单独的蛋白质- dna复合物。
Single-molecule studies of protein–DNA interactions continue to yield new information on numerous DNA processing pathways. For example, optical microscopy-based techniques permit the real-time observation of proteins that interact with DNA substrates, which in turn allows direct insight into reaction mechanisms. However, these experiments remain technically challenging and are limited by the paucity of stable chromophores and the difficulty of acquiring statistically significant observations. In this protocol, we describe a novel, high-throughput, nanofabricated experimental platform enabling real-time imaging of hundreds of individual protein–DNA complexes over hour timescales.