MICROFLUIDS, MICROREACTIONS, AND MICROSEPARATIONS

微流体、微反应和微分离

• 批准号: 6442988
• 负责人: Mark A BURNS
• 金额: $ 16.41万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6442988
• 关键词: DNA; biochemistry; biomedical equipment development; computer program /software; evaluation /testing; fluid; fluid flow; fluorescent dye /probe; gel electrophoresis; genotype; method development; microinjections; miniature biomedical equipment; nucleic acid quantitation /detection; nucleic acid sequence

Analysis methods for DNA genotyping and sequencing are linked into multiple-step processing systems that begin with DNA source material and extract genetic information. In conventional methods, the processing system operates on batches of microliter-volume reaction samples, together with matched liquid handling devices and electrophoretic analysis equipment. Project 2 will provide component-level design and testing our improved nanoliter-volume processing components for DNA sequencing and genotyping. To fulfill the goal of performing complex multi-step tasks on a single device, discrete drop sample handling and informational control will be required. New or improved fabrication technologies will also be examined as needed. All of the components developed in the Project retain the target of eventual integration in the complex systems of Project 1. Project 2 has three Specific Aims. 1) Develop a nanoliter injection and movement system. We will construct and test components for a robust and flexible injection and movement system that can handle multiple liquid samples in the volume range of 10 nl to 500 nl. Our successful injection and movement systems for single and binary drop systems will be modified and assembled into multiple- sample liquid handling procedures. Each discrete drop will be addressed and manipulated independently. 2) Develop multiple independent reaction chambers. We will construct and test reaction chambers for optimal surface interactions, temperature profiles, and thermal cross talk between reaction chamber components. A significant effort will involve developing control software and reproducible reaction conditions for high quality DNA sequencing. 3) Demonstrate high-resolution separations. We will improve the existing gel electrophoresis system (approximately 10-50 bp resolution, 500 bp length), to approach the quality of conventional large-scale DNA sequencing (1 bp resolution, 1000 bp length). Optimization of the existing fluorescent labeling and detection methods will be emphasized. Additional effort will examine gel electrophoresis matrix modifications and the effect of alternative processing biochemistries.

DNA基因分型和测序的分析方法链接到多步处理系统，从DNA源材料开始并提取遗传信息。在常规方法中，加工系统与微量 - 体积反应样品的批处理以及匹配的液体处理设备和电泳分析设备一起运行。项目2将提供组件级的设计，并测试我们改进的纳米素体积处理组件，用于DNA测序和基因分型。为了实现在单个设备上执行复杂的多步任务的目标，将需要离散的删除示例处理和信息控制。还将根据需要检查新的或改进的制造技术。项目中开发的所有组件都保留了项目1的复杂系统中最终集成的目标。项目2具有三个具体目标。 1）开发纳米仪注入和运动系统。我们将构建和测试组件，以稳健而柔性的注入和运动系统，该系统可以处理10 nl至500 nl的体积范围内的多个液体样品。我们成功的单次和二进制液滴系统的注入和移动系统将被修改并组装成多样品液体处理程序。每个离散的下降将被解决并独立操纵。 2）发展多个独立反应室。我们将构建和测试反应室，以在反应室成分之间进行最佳的表面相互作用，温度曲线和热横聊。一项巨大的努力将涉及开发控制软件和可再现的反应条件，以进行高质量的DNA测序。 3）展示高分辨率分离。我们将改善现有的凝胶电泳系统（大约10-50 bp的分辨率，500 bp的长度），以接近常规大规模DNA测序的质量（1 bp分辨率，1000 bp长度）。将强调现有荧光标签和检测方法的优化。额外的努力将检查凝胶电泳基质修饰以及替代加工生物学的影响。