A dual-beam optical microscope for observation and cleavage of single DNA molecules.
A dual-beam optical microscope for observation and cleavage of single DNA molecules.
复制标题
用于观察和切割单个 DNA 分子的双光束光学显微镜。
作者：
W. A. Lyon;M. Fang;W. Haskins;S. Nie
集成的外焦和倏逝波光学显微镜已被开发出来，用于单个 DNA 分子的实时观察和键断裂研究。大的基因组 DNA 在层流中拉伸，并通过强静电相互作用固定在聚赖氨酸涂层的玻璃表面上。与之前的单分子动力学和单酶研究不同，这项工作利用双链 DNA 的弹性性质，测量由两个磷酸二酯 (P-O) 键断裂触发的 DNA 弛豫事件。追踪单个 DNA 分子的化学反应的能力为 DNA 作图和研究 DNA-蛋白质相互作用开辟了新的可能性。
An integrated epifocal and evanescent-wave optical microscope has been developed for real-time observation and bond cleavage studies of single DNA molecules. Large genomic DNA is stretched in a laminar flow stream and is immobilized on a polylysine-coated glass surface by strong electrostatic interactions. Unlike previous single-molecule dynamics and single-enzyme studies, this work takes advantage of the elastic nature of double-stranded DNA and measures DNA relaxation events that are triggered by two phosphodiester (P-O) bond breaks. The ability to follow chemical reactions on individual DNA molecules opens new possibilities for DNA mapping and for studying DNA-protein interactions.