Microdevice for Direct DNA Purification

用于直接 DNA 纯化的微型装置

• 批准号: 7498330
• 负责人: James P Landers
• 金额: $ 6.45万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-19 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7498330
• 关键词: Area; Arts; Binding; Biological; Blood; Blood Cells; Buffers; Businesses; Cancer Biology; Carbohydrates; Cells; Charge; Chemistry; Compatible; Complex; Coupled; Couples; Coupling; Cytolysis; DNA; DNA purification; Development; Devices; Diagnosis; Ethylene; Ethylenes; Freezing; Gel; Gene Mutation; Glass; Goals; Histologic; Hybrids; Individual; Industry; Lasers; Malignant Neoplasms; Membrane; Methodology; Methods; Microfabrication; Microfluidic Microchips; Modification; Molecular; Performance; Phase; Plastics; Polymerase Chain Reaction; Polymers; Preparation; Process; Research; Sampling; Silicon Dioxide; Solid; Source; Standards of Weights and Measures; Structure; Surface; Surgical Pathology; System; Techniques; Testing; Time; Tissue Sample; Tissues; Universities; Virginia; anticancer research; base; cancer cell; cell type; cost; design; genetic analysis; instrument; interest; laser capture microdissection; lithography; microchip; multidisciplinary; skills; vinyl acetate

With an ever-increasing interest in the molecular typing of cells from histologic tissue sections, the ability rapidly and efficiently extract DNA from the selected cells will be of paramount importance. The overall goal of this project is to develop a microchip-based sample preparation method for high efficiency, low-cost extraction of DNAfrom tissue samples - this microdevice will easily accommodate blood or other cells sources. The microdevices will be created using state-of-the-art microfabrication techniques coupled with fluidically-controlled on-chip cell lysis and solid phase extraction chemistries. This project couples the industrial capabilities of HT Micro for facile fabrication of complex, high surface area microstructures, with surface modification chemistries developed at the University of Virginia that enable efficient and high capacity DNA extraction. The microdevices will be tested using a variety of sample varying in type and quality, and extraction efficiency will be determined using real-time PCR. As a demonstration of integration with current laser-capture microdissection instruments, the microdevice will be fabricated to directly accept the cap from the Arcturus Pixcell lie which will contain the selected cells bound to an ethylene vinyl acetate polymer membrane on its bottom surface.. The tissue samples will be collected and laser microdissected by our surgical pathology collaborator here at UVa - samples of normal and malignant cells will be analyzed. The final device will offer the rapid analysis, high extraction efficiency and high-throughput advantages of microdevices and, in addition, is expected to offer higher capacity, and lower cost-per-device than current conventional or microchip techniques.

随着对来自组织学组织切片的细胞的分子分型的兴趣不断增加， 从选择的细胞中快速有效地提取DNA将是至关重要的。总目标 本课题的主要目的是开发一种基于微芯片的高效、低成本的样品前处理方法 从组织样本中提取DNA-这种微型装置将很容易容纳血液或其他细胞 源微器件将使用最先进的微制造技术， 流体控制的芯片上细胞裂解和固相萃取化学。该项目将 HT Micro的工业能力，可轻松制造复杂的高表面积微结构， 弗吉尼亚大学开发的表面改性化学技术， DNA提取能力将使用各种类型的样品对微型器件进行测试， 质量和提取效率将使用实时PCR确定。作为融合的示范 使用当前的激光捕获显微切割仪器， 来自大角星Pixcell Lie的帽子将包含与乙烯乙酸乙烯酯结合的选定细胞 聚合物膜在其底部表面上。将收集组织样本，并通过激光显微切割 我们在UVa的外科病理学合作者将分析正常和恶性细胞的样本。 最终的设备将提供快速分析，高提取效率和高通量的优势， 此外，它还有望提供比目前更高的容量和更低的每设备成本 常规或微芯片技术。