期刊
LANGMUIR
卷 24, 期 19, 页码 11293-11299出版社
AMER CHEMICAL SOC
DOI: 10.1021/la8017634
关键词
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资金
- Initiatives in Science and Engineering (ISE)
- NIH [GM074739]
- NSF PECASE
- NSF Graduate Research fellowship
- Nanoscale Science and Engineering Initiative of the National Science Foundation [CHE-0641523]
- New York State Office of Science, Technology, and Academic Research (NYSTAR)
The analysis of individual molecules is evolving into an important tool for biological research, and presents conceptually new ways of approaching experimental design strategies. However, more robust methods are required if these technologies are to be made broadly available to the biological research community. To help achieve this goal we have combined nanofabrication techniques with single-molecule optical microscopy for assembling and visualizing curtains comprised of thousands of individual DNA molecules organized at engineered diffusion barriers on a lipid bilayer-coated surface. Here we present an important extension of this technology that implements geometric barrier patterns comprised of thousands of nanoscale wells that can be loaded with single molecules of DNA. We show that these geometric nanowells can be used to precisely control the lateral distribution of the individual DNA molecules within curtains assembled along the edges of the engineered barrier patterns. The individual molecules making up the DNA curtain can be separated from one another by a user-defined distance dictated by the dimensions of the nanowells. We demonstrate the broader utility of these patterned DNA curtains in a novel, real time restriction assay that we refer to as dynamic optical restriction mapping, which can be used to rapidly identify entire sets of cleavage sites within a large DNA molecule.
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