人类细胞中核酸的细胞内核磁共振光谱

Q4 Chemistry Current Protocols in Nucleic Acid Chemistry Pub Date : 2018-11-29 DOI:10.1002/cpnc.71

Pavlina Viskova, Daniel Krafcik, Lukas Trantirek, Silvie Foldynova-Trantirkova

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引用次数: 7

摘要

细胞内核磁共振波谱是一种独特的工具，可以研究生物分子的结构和动力学，以及它们在接近原子分辨率的活细胞复杂环境中的相互作用。在本文中，详细说明了建立一个细胞内核磁共振实验，以监测DNA寡核苷酸的结构，通过量身定制的电穿孔引入到活的人类细胞的细胞核。详细的一步一步的协议，无论是在细胞内核磁共振样品的制备，以及协议进行必要的控制实验，包括流式细胞术和共聚焦显微镜描述。讨论了细胞内核磁共振实验的优点和局限性。©2018 by John Wiley &儿子,Inc。

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In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells

In-cell NMR spectroscopy is a unique tool that enables the study of the structure and dynamics of biomolecules as well as their interactions in the complex environment of living cells at near-to-atomic resolution. In this article, detailed instructions are described for setting up an in-cell NMR experiment for monitoring structures of DNA oligonucleotides introduced into nuclei of living human cells via tailored electroporation. Detailed step-by-step protocols for both the preparation of an in-cell NMR sample as well as protocols for conducting essential control experiments including flow cytometry and confocal microscopy are described. The strengths and limitations of in-cell NMR experiments are discussed. © 2018 by John Wiley & Sons, Inc.

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Current Protocols in Nucleic Acid Chemistry Chemistry-Organic Chemistry

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期刊介绍： Published in association with International Society for Nucleosides, Nucleotides & Nucleic Acids (IS3NA) , Current Protocols in Nucleic Acid Chemistry is equally valuable for biotech, pharmaceutical, and academic labs. It is the resource for designing and running successful research projects in the rapidly growing and changing field of nucleic acid, nucleotide, and nucleoside research.

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