Single Molecule Imaging of DNA-Protein Interactions Using DNA Curtains.
Single Molecule Imaging of DNA-Protein Interactions Using DNA Curtains.
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DOI：
10.1007/978-1-0716-2847-8_10
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发表时间：
2023
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直接观察促进核酸代谢的酶是了解其生化和生物学特性的有效方法。多年来，荧光光学显微镜已发展成为实时观察生物路径的强大工具。在这里，我们将 DNA 幕帘描述为一种光学显微镜工具，它结合了工程、生物化学和单分子成像，可以实时直接观察酶对 DNA 的作用。我们将提供这种方法的详细方法，包括有关基本 TIRF 显微镜的设置、用于成像的流动室组装以及制作 DNA 帘的协议的信息。我们的目标是帮助读者更好地理解DNA窗帘的技术方法，并更好地理解这种方法的生化和生物学应用。
Direct observation of enzymes that work to promote nucleic acid metabolism is a powerful approach to understanding their biochemical and biological properties. Over several years, fluorescent optical microscopy has developed as a powerful tool for watching biological pathways as they occur in real time. Here we describe DNA curtains as an optical microscopy tool that combines engineering, biochemistry, and single molecule imaging to make direct observations of enzymes as they work on DNA in real time. We will provide a detailed methodology of this approach including information about the setup of a basic TIRF microscope, assembly of flow chambers for imaging, and the protocol for making DNA curtains. Our goal is to help the reader better understand the technical approaches to DNA curtains and to better understand the biochemical and biological applications of this approach.