MICROFLUIDS, MICROREACTIONS, AND MICROSEPARATIONS

微流体、微反应和微分离

• 批准号: 6318373
• 负责人: Mark A BURNS
• 金额: $ 37.64万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-05 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6318373
• 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被连接到从DNA来源材料开始并提取遗传信息的多步骤处理系统中。在传统方法中，处理系统处理一批微升体积的反应样品，以及匹配的液体处理设备和电泳分析设备。项目2将为DNA测序和基因分型提供组件级设计和测试我们改进的纳升体积处理组件。为了实现在单个设备上执行复杂的多步骤任务的目标，将需要离散的液滴样品处理和信息控制。新的或改进的制造技术也将根据需要进行审查。项目中开发的所有组件都保留了最终集成到项目1的复杂系统中的目标。项目2有三个具体目标。1)研制了纳升注射和移动系统。我们将为一个坚固而灵活的注射和移动系统建造和测试部件，该系统可以处理体积在10 NL到500 NL范围内的多种液体样品。我们成功的单液滴和双液滴系统的注入和移动系统将进行改进，并组装成多样品液体处理程序。每个离散的液滴都将被单独处理和操作。2)开发多个独立的反应室。我们将建造和测试反应室，以获得最佳的表面相互作用、温度分布和反应室组件之间的热串扰。一项重要的工作将涉及开发控制软件和可重现的反应条件，以实现高质量的DNA测序。3)演示高分辨率分色。我们将改进现有的凝胶电泳系统(分辨率约为10-50个碱基，长度约为500个碱基)，以接近常规大规模DNA测序(分辨率为1个碱基，长度为1000个碱基)的质量。重点是优化现有的荧光标记和检测方法。额外的工作将检查凝胶电泳基质修饰和替代工艺生物化学的效果。