Visualization of DNA and protein-DNA complexes with atomic force microscopy.
Visualization of DNA and protein-DNA complexes with atomic force microscopy.
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与原子力显微镜的DNA和蛋白-DNA复合物的可视化。
DOI：
10.1007/978-1-62703-776-1_17
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发表时间：
2014
期刊：
影响因子：
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通讯作者：
Shlyakhtenko, Luda S
中科院分区：
文献类型：
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作者：
Lyubchenko, Yuri L;Gall, Alexander A;Shlyakhtenko, Luda S
本文介绍了DNA和蛋白质-DNA复合物的AFM成像的样品制备技术。该方法是基于化学官能化的云母表面与氨丙基杂氮硅三环（APS），以产生APS-云母表面。该表面结合核酸和核蛋白复合物在广泛的离子强度范围内，在没有二价阳离子，并在广泛的pH值范围内。本章介绍了APS-云母表面的制备方法和AFM成像样品的制备。并给出了APS的合成和纯化方法。原子力显微镜的应用说明与DNA和蛋白质-DNA复合物的图像的例子。
This article describes sample preparation techniques for AFM imaging of DNA and protein–DNA complexes. The approach is based on chemical functionalization of the mica surface with aminopropyl silatrane (APS) to yield an APS-mica surface. This surface binds nucleic acids and nucleoprotein complexes in a wide range of ionic strengths, in the absence of divalent cations, and in a broad range of pH. The chapter describes the methodologies for the preparation of APS-mica surfaces and the preparation of samples for AFM imaging. The protocol for synthesis and purifi cation of APS is also provided. The AFM applications are illustrated with examples of images of DNA and protein–DNA complexes.