Lyubchenko Yuri L, Gall Alexander A, Shlyakhtenko Luda S
Department of Pharmaceutical Sciences, University of Nebraska Medical Center, Omaha, NE, USA.
Methods Mol Biol. 2014;1117:367-84. doi: 10.1007/978-1-62703-776-1_17.
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 purification of APS is also provided. The AFM applications are illustrated with examples of images of DNA and protein-DNA complexes.
本文介绍了用于DNA和蛋白质-DNA复合物原子力显微镜（AFM）成像的样品制备技术。该方法基于用氨丙基硅氮烷（APS）对云母表面进行化学功能化，以产生APS-云母表面。该表面能在广泛的离子强度范围内、在不存在二价阳离子的情况下以及在较宽的pH范围内结合核酸和核蛋白复合物。本章描述了APS-云母表面的制备方法以及用于AFM成像的样品制备方法。还提供了APS的合成和纯化方案。通过DNA和蛋白质-DNA复合物的图像示例展示了AFM的应用。
