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在玻璃基底上简单拉伸 DNA 的方法,用于荧光成像和光谱学研究。

Simple method of DNA stretching on glass substrate for fluorescence imaging and spectroscopy.

机构信息

IBS Center for Integrated Nanostructure Physics, Institute for Basic Science (IBS), Daejeon 305-701, Republic of KoreabSungkyunkwan University, Department of Energy Science, Suwon 440-746, Republic of Korea.

IBS Center for Integrated Nanostructure Physics, Institute for Basic Science (IBS), Daejeon 305-701, Republic of Korea.

出版信息

J Biomed Opt. 2014 May;19(5):051210. doi: 10.1117/1.JBO.19.5.051210.

Abstract

We demonstrate a simple method of stretching DNA to its full length, suitable for optical imaging and atomic force microscopy (AFM). Two competing forces on the DNA molecules, which are the electrostatic attraction between positively charged dye molecules (YOYO-1) intercalated into DNA and the negatively charged surface of glass substrate, and the centrifugal force of the rotating substrate, are mainly responsible for the effective stretching and the dispersion of single strands of DNA. The density of stretched DNA molecules could be controlled by the concentration of the dye-stained DNA solution. Stretching of single DNA molecules was confirmed by AFM imaging and the photoluminescence spectra of single DNA molecule stained with YOYO-1 were obtained, suggesting that our method is useful for spectroscopic analysis of DNA at the single molecule level.

摘要

我们展示了一种将 DNA 拉伸至全长的简单方法,适用于光学成像和原子力显微镜(AFM)。两种相互竞争的力作用于 DNA 分子,一种是嵌入 DNA 的带正电荷的染料分子(YOYO-1)之间的静电吸引力,另一种是带负电荷的玻璃基底的表面力,以及旋转基底的离心力,它们主要负责有效地拉伸和分散 DNA 的单链。拉伸 DNA 分子的密度可以通过染色 DNA 溶液的浓度来控制。通过 AFM 成像证实了单 DNA 分子的拉伸,并且获得了用 YOYO-1 染色的单个 DNA 分子的光致发光光谱,表明我们的方法可用于单分子水平的 DNA 光谱分析。

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