Spin-stretching of DNA and protein molecules for detection by fluorescence and atomic force microscopy
Spin-stretching of DNA and protein molecules for detection by fluorescence and atomic force microscopy
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DOI：
10.1021/ac9902695
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发表时间：
1999-10-01
影响因子：
7.4
通讯作者：
Aebersold, R
中科院分区：
文献类型：
--
作者：
Yokota, H;Sunwoo, J;Aebersold, R
我们已经开发出一种快速有效的方法来拉伸DNA和变性蛋白质分子，用于荧光显微镜和原子力显微镜（AFM）的检测。在所描述的方法中，由瞬时旋转产生的粘性阻力现在拉伸随机卷曲的DNA分子或变性蛋白质。通过将含有DNA或变性蛋白质的样品溶液液滴分配到MgCl2浸泡的云母表面上来实现拉伸。我们目前的荧光图像拉直rDNA分子和AFM图像的应力剪切，减少冯维勒布兰德因子以及拉直rDNA。所描述的快速和可靠的自旋拉伸技术将在单个生物聚合物分子的分析中找到广泛的应用。
We have developed a rapid and efficient way of stretching DNA and denatured protein molecules for detection by fluorescence microscopy and atomic force microscopy (AFM). In the described method, a viscous drag created by a transient rotational now stretches randomly coiled DNA molecules or denatured proteins. Stretching is achieved by dispensing a droplet of sample solution containing DNA or denatured protein on a MgCl2-soaked mica surface. We present fluorescent images of straightened RDNA molecules and AFM images of stress-sheared, reduced von Willebrand factor as well as straightened RDNA. The described quick and reliable spin-stretching technique will find wide applications in the analysis of single biopolymer molecules.