CAS OpenIR  > 中科院上海应用物理研究所2004-2010年
Alternative TitleTowards molecular surgery with nano-manipulation of single DNA molecules
Thesis Advisor胡钧
Degree Grantor中国科学院上海应用物理研究所
Place of Conferral中国科学院上海应用物理研究所
Degree Discipline无机化学
Keyword分子手术 原子力显微镜 Dna单分子 纳米操纵 定位拾取 测序策略
Other Abstract"Molecular surgery" means " modifying biomolecules with nanometer resolution at the single molecule level in the principle of optics, mechanics or enzymology". Our motivation is to realize precise manipulation of single DNA molecules using atomic force microscopy, to develop some key techniques, and to explore its possible applications in biology. Some issues involved in "operating table" were figured out. After further purification of the DNA molecules, substrates for DNA manipulation were prepared by modifying the mica with nickel ion or further treating the 3-aminopropyl triethoxysilane modified mica under a temperature of 120 ℃ for more than 4 hours. To match the fact that DNA should be weakly deposited on the solid surface, centrifugal method was used to stretched long strands of A DNA as well as pBR322 DNA. A unique technique for the positioning dissection and isolation of single DNA molecules by using atomic force microscopy (AFM) has been demonstrated. Based on lift mode in the interleave scan, single DNA fragment was dissected from one of that genome DNA strands with the AFM tip at the desired position, and then isolated (or picked up) after a special operation called "kneading". All the operations including imaging, dissection and isolation could be carried out with one tip. The isolated DNA fragment on the AFM tip could be successfully amplified by single molecule PCR. Some of the physical and chemical mechanisms have been discussed. A new strategy for DNA sequencing in order was proposed. Ordered sequencing strategy. Combined with traditional sequencing methods based on gel electrophoresis, the nanomanipulation technique of single DNA molecules was applied to sequencing the genome sequences. Two hybrid strategies were also brought forward. Compared to traditional sequencing methods, our strategy has some obviously advantages, such as less time and cost, more simple, apt to assembly, and so on.
Document Type学位论文
Recommended Citation
GB/T 7714
吕军鸿. 面向分子手术的DNA单分子的纳米操纵研究[D]. 中国科学院上海应用物理研究所. 中国科学院上海应用物理研究所,2004.
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