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DNA的原子力显微镜应用。

Applications for atomic force microscopy of DNA.

作者信息

Hansma H G, Laney D E, Bezanilla M, Sinsheimer R L, Hansma P K

机构信息

Department of Physics, University of California, Santa Barbara 93106, USA.

出版信息

Biophys J. 1995 May;68(5):1672-7. doi: 10.1016/S0006-3495(95)80343-7.

Abstract

Tapping mode atomic force microscopy (AFM) of DNA in propanol, dry helium, and aqueous buffer each have specific applications. Resolution is best in propanol, which precipitates and immobilizes the DNA and provides a fluid imaging environment where adhesive forces are minimized. Resolution on exceptional images of DNA appears to be approximately 2 nm, sufficient to see helix turns in detail, but the smallest substructures typically seen on DNA in propanol are approximately 6-10 nm in size. Tapping AFM in dry helium provides a convenient way of imaging such things as conformations of DNA molecules and positions of proteins on DNA. Images of single-stranded DNA and RecA-DNA complexes are presented. In aqueous buffer DNA molecules as small as 300 bp have been imaged even when in motion. Images are presented of the changes in shape and position of circular plasmid DNA molecules.

摘要

在丙醇、干燥氦气和水性缓冲液中对DNA进行轻敲模式原子力显微镜(AFM)成像各有特定应用。在丙醇中分辨率最佳,丙醇能使DNA沉淀并固定,提供一个使粘附力最小化的流体成像环境。DNA特殊图像上的分辨率似乎约为2纳米,足以详细观察到螺旋结构,但在丙醇中DNA上通常能看到的最小亚结构大小约为6 - 10纳米。在干燥氦气中进行轻敲AFM成像为观察DNA分子构象和蛋白质在DNA上的位置等提供了一种便捷方法。展示了单链DNA和RecA - DNA复合物的图像。即使在运动状态下,小至300碱基对的DNA分子在水性缓冲液中也已成像。展示了环状质粒DNA分子形状和位置变化的图像。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db5d/1282069/122c17b00559/biophysj00061-0039-a.jpg

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