CAREER: Dynamics of DNA during Electrophoresis in Artificial Sieving Matrices

职业：人工筛分基质中电泳过程中 DNA 的动力学

• 批准号: 0642794
• 负责人: Kevin Dorfman
• 金额: $ 40万
• 依托单位: University of Minnesota-Twin Cities
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0642794&HistoricalAwards=false
• 关键词: CAREER; Dynamics; DNA; during; Electrophoresis

Proposal Number CTS-0642794Principal Investigator: Kevin D. DorfmanUniversity/Institution: University of Minnesota-Twin Cities CAREER: Dynamics of DNA during Electrophoresis in Artificial Sieving MatricesThis project involves theoretical modeling and experimental investigation of microfluidic electrophoresis of DNA. DNA electrophoresis remains not only one of the most important tools in molecular biology and genetics, but also an ideal platform for testing fundamental theories of polymer dynamics and coarse-grained averaging of separation processes. The proposed research will address topics relevant to polymer physics, stochastic processes, and chemical engineering, working across multiple length scales to connect microscopic motion to macroscopic observables. The research component is an integrated theoretical and experimental program that addresses a range of fundamental issues arising during DNA separations in microfabricated post arrays, including the hindered relaxation phenomena in an external field and the role of the order of the post lattice. The research focuses on effects that arise from the proximity of posts in real separation devices, which are not addressed by the existing body of knowledge on DNA collisions with a single, isolated post. The experimental work spans multiple scales, combining ensemble-averaged measurements of the transport over several millimeters with detailed single-molecule observations of the local transport at the macromolecular scale. The microscopic insights will be used to construct new theoretical models, based upon continuous-time random walk theory, that rationalize the ensemble-averaged experimental results and predict new behavior that can be tested experimentally. In addition to training graduate students, the education component of this proposal consists of three parts: (i) developing a new course on the theory of electrophoresis and lab-on-a-chip technology; (ii) summer research opportunities for underrepresented minorities through a collaborative outreach program with the University of Puerto Rico at Mayaguez; and (iii) a K-12 outreach program with Edison High School in Minneapolis.Most importantly, the research will link theory to experiment in a well-defined microstructured environment. The proposed work will lead to improved DNA separation devices, suitable for applied and fundamental research. Artificial matrices are poised to replace pulsed-field gel electrophoresis for important applications in genome mapping and DNA fingerprinting, but the incomplete understanding of the dynamics of DNA in artificial matrices has prevented this technique from moving beyond the proof-of-principle stage and into routine use. The fundamental results could also impact related separation techniques, such as electrochromatography and microfluidic HPLC. The summer research program with the University of Puerto Rico at Mayaguez will provide advanced training in computational and experimental techniques that are unavailable to these students at their home university. The outreach program with Edison High School in Minneapolis will integrate the research activities with education at the K-12 level.

提议编号 CTS-0642794主要研究者：Kevin D. Dorfman大学/机构：明尼苏达大学双城分校职业：人工筛分基质电泳过程中DNA的动力学本项目涉及DNA微流控电泳的理论建模和实验研究。DNA电泳不仅是分子生物学和遗传学中最重要的工具之一，也是测试聚合物动力学和分离过程粗粒度平均的基础理论的理想平台。拟议的研究将涉及与聚合物物理学，随机过程和化学工程相关的主题，跨多个长度尺度工作，将微观运动与宏观可观测量联系起来。 该研究部分是一个综合的理论和实验计划，解决了一系列的基本问题，在DNA分离过程中产生的微制造后阵列，包括在外部领域的阻碍弛豫现象和后晶格的顺序的作用。该研究的重点是在真实的分离装置中由柱的接近引起的效应，这是现有的关于DNA与单个孤立的柱碰撞的知识体系所不能解决的。实验工作跨越多个尺度，将几毫米内的整体平均传输测量与大分子尺度下局部传输的详细单分子观测相结合。微观见解将用于构建新的理论模型，基于连续时间随机游走理论，合理化整体平均实验结果，并预测可以通过实验测试的新行为。除了培训研究生外，这项建议的教育部分包括三个部分：㈠编写关于电泳理论和芯片实验室技术的新课程; ㈡通过与马亚圭斯的波多黎各大学的合作推广方案，为代表人数不足的少数群体提供夏季研究机会;以及（iii）与明尼阿波利斯爱迪生高中的K-12外展计划。最重要的是，该研究将在定义明确的微结构环境中将理论与实验联系起来。 拟议的工作将导致改进的DNA分离装置，适用于应用和基础研究。人工基质有望取代脉冲场凝胶电泳在基因组作图和DNA指纹图谱中的重要应用，但对人工基质中DNA动力学的不完全理解阻止了该技术超越原理验证阶段并进入常规使用。基本结果也可能影响相关的分离技术，如电色谱和微流控HPLC。与位于马亚圭斯的波多黎各大学的夏季研究方案将提供这些学生在其家乡大学无法获得的计算和实验技术方面的高级培训。与明尼阿波利斯爱迪生高中的外联计划将把研究活动与K-12水平的教育结合起来。