Soft matter on-a-chip: Nanofluidics for single molecule DNA analysis and manipulation

软物质芯片：用于单分子 DNA 分析和操作的纳米流体

• 批准号: 386212-2010
• 负责人: Reisner, Walter
• 金额: $ 2.33万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2017
• 资助国家: 加拿大
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=633495
• 关键词: Soft; matter; chip; Nanofluidics; single

Nanofluidic devices are networks of fluid-filled channels etched on a chip with dimensions on order of 1-100nm. Devices with such small dimensions can have radically different properties than their macro world analogs, in particular the ability to manipulate and analyze single molecules. The core of the PI's research program-both pure and applied-is the use of nanofluidic devices for the manipulation and analysis of single molecules of DNA. DNA is of fundamental importance to biology, an important nanoscale structural material in its own right and a model system for understanding the physics of polyelectrolytes in confinement. The goal of the PI's fundamental research program is to create new knowledge (new scaling laws, explore new phenomena) while translating the basic science into new device concepts. The applied component makes use of nanofluidic technology to tackle immediate problems in biomedicine. When DNA is squeezed into a small 'nanopipe' (e.g. using high pressure) the molecule will stretch out, unscrolling the genome along the channel. The PI has developed a new 'melting mapping' approach for analyzing the unscrolled DNA. The extended DNA is partially melted via chemical and thermal denaturation, resulting in the co-existence of double stranded and single stranded regions along the molecule. This melting pattern is sequence dependent. By using dyes which unbind from melted regions, the local melting pattern can be observed optically as a "bar-code" of alternating dark and bright regions. The long term goal is to use this barcoding technology to create maps of entire genome to aid in sequencing, the study of large-scale structural variation and as a tool for single cell genomic analysis.

纳米流体装置是蚀刻在芯片上的流体填充通道的网络，其尺寸为1- 100 nm。具有如此小尺寸的设备可以具有与其宏观世界类似物完全不同的特性，特别是操纵和分析单个分子的能力。PI的研究计划的核心-无论是纯的还是应用的-是使用纳米流体设备来操纵和分析DNA的单分子。DNA对生物学至关重要，它本身就是一种重要的纳米结构材料，也是理解聚电解质物理学的模型系统。PI的基础研究计划的目标是创造新的知识（新的比例定律，探索新的现象），同时将基础科学转化为新的设备概念。应用组件利用纳米流体技术来解决生物医学中的紧迫问题。当DNA被挤压到一个小的“纳米管”（例如使用高压）中时，分子将伸展，基因组沿着通道展开。PI开发了一种新的“熔解映射”方法来分析解卷的DNA。延伸的DNA通过化学和热变性而部分解链，导致双链和单链区域沿分子沿着共存。这种解链模式是序列依赖性的。通过使用从熔化区域解结合的染料，可以光学地观察到局部熔化图案作为交替的暗区域和亮区域的“条形码”。长期目标是使用这种条形码技术创建整个基因组的图谱，以帮助测序，研究大规模结构变异，并作为单细胞基因组分析的工具。