Enhanced Resolution in Microdevice Electrophoresis

提高微型器件电泳的分辨率

• 批准号: 6740088
• 负责人: Victor M Ugaz
• 金额: $ 15.82万
• 依托单位: TEXAS ENGINEERING EXPERIMENT STATION
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6740088
• 关键词: DNA; biomedical equipment development; biotechnology; electrophoresis; miniature biomedical equipment; nanotechnology; nucleic acid quantitation /detection; nucleic acid sequence; technology /technique development

DESCRIPTION (provided by applicant): The DNA sequence data generated by the Human Genome Project continues to revolutionize the field of medicine by uncovering the genetic basis for many diseases. Unfortunately, the considerable expense associated with the purchase and operation of specialized equipment and the requirement of a dedicated laboratory infrastructure staffed by highly trained personnel currently renders sequencing impractical for use in routine medical practice. Microfabricated sequencing devices are poised to offer an inexpensive alternative to conventional equipment, ultimately making direct genomic sequencing practical for use as a true point-of-care diagnostic protocol. It addition, moving to the microdevice format substantially reduces the sample volumes required for analysis (to the order of nanoliters) and introduces the ability to fabricate self-contained portable sequencing devices which can be easily deployed in remote field locations for a variety of applications, including the rapid identification and tracking of infectious diseases. A key element of any sequencing device is the ability to perform size-based separations of DNA fragments using electrophoresis. Complete sequencing requires sufficient resolution to distinguish fragments which differ by only one base pair in length in fragments approaching 1,000 base pairs in total length - a requirement that is routinely met by conventional slab gel and capillary electrophoresis systems. At the present time, separation channel lengths on the order of several centimeters are required to achieve single base pair resolution in microfabricated electrophoresis systems. Though impressive, the size of current generation microfabricated sequencing devices is still too large to exploit more than a fraction of the enormous cost savings possible through mass production via photolithographic fabrication techniques. These techniques, routinely employed in the semiconductor industry, allow tens or hundreds of devices to be produced on a single wafer. The specific aim of this research project is to demonstrate that microdevice-based electrophoretic separations can achieve resolution comparable to conventional macro-scale DNA analysis systems over length scales on the order of one centimeter or less. The Genome Scholar Development and Faculty Transition Award will provide the opportunity to obtain training in the key areas of microfabrication techniques and molecular biology methods in order to meet this research objective.

描述（由申请人提供）： 人类基因组计划继续彻底改变医学领域， 揭示了许多疾病的遗传基础。不幸的是， 与购买和操作专用设备有关的费用， 需要一个专门的实验室基础设施， 受过训练的人员目前使得测序不适用于常规测序。 医疗实践。微制造的测序设备准备提供一个 传统设备的廉价替代品， 可用作真正的即时诊断的基因组测序 议定书此外，转向微型设备格式大大减少了 分析所需的样品体积（纳升数量级），以及 介绍了制造独立便携式测序设备的能力 它可以很容易地部署在远程现场位置， 应用，包括快速识别和跟踪传染病 疾病 任何测序设备的关键要素是执行基于大小的测序的能力。 使用电泳分离DNA片段。完全测序 需要足够的分辨率来区分仅相差 在总共接近1，000个碱基对的片段中长度为一个碱基对 长度-常规平板凝胶通常满足要求， 毛细管电泳系统目前，分离通道 需要几厘米数量级的长度来实现单碱基 微加工电泳系统中的对分辨率。虽然令人印象深刻， 当前一代微制造测序装置的尺寸仍然太大 大到利用超过一小部分的巨大成本节省可能 通过可编程制造技术进行大规模生产。这些 半导体工业中常规采用的技术允许数十或 在一个晶片上生产数百个器件。其具体目的是 研究项目是证明基于微器件的电泳 分离可以达到与常规宏观尺度DNA相当的分辨率， 分析系统的长度尺度在一厘米或更小的数量级上。 基因组学者发展和教师过渡奖将提供 获得微细加工技术关键领域培训的机会 和分子生物学方法，以满足这一研究目标。