A Dual-Beam Optical Microscope for Observation and Cleavage of Single DNA Molecules

详细信息    查看全文

• 作者：William A. Lyon ; Michelle M. Fang ; William E. Haskins ; and Shuming Nie
• 刊名：Analytical Chemistry
• 出版年：1998
• 出版时间：May 1, 1998
• 年：1998
• 卷：70
• 期：9
• 页码：1743 - 1748
• 全文大小：215K
• 年卷期：v.70,no.9(May 1, 1998)
• ISSN：1520-6882

文摘

An integrated epifocal and evanescent-wave optical microscope has been developed for real-time observationand bond cleavage studies of single DNA molecules.Large genomic DNA is stretched in a laminar flowstreamand is immobilized on a polylysine-coated glass surfaceby strong electrostatic interactions. Unlike previoussingle-molecule dynamics and single-enzyme studies, thiswork takes advantage of the elastic nature of double-stranded DNA and measures DNA relaxation events thatare triggered by two phosphodiester (P-O) bond breaks.The ability to follow chemical reactions on individualDNAmolecules opens new possibilities for DNA mapping andfor studying DNA-protein interactions.