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Enhanced Resolution in Microdevice Electrophoresis

提高微型器件电泳的分辨率

基本信息

批准号：
6922879
负责人：
Victor M Ugaz
金额：
$ 26.19万
依托单位：
TEXAS ENGINEERING EXPERIMENT STATION
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-09-01 至 2007-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6922879
关键词：
DNA biomedical equipment development biotechnology electrophoresis miniature biomedical equipment nanotechnology nucleic acid quantitation /detection nucleic acid sequence technology /technique development

项目摘要

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)：由人类基因组计划继续为医学领域带来革命性的变化揭示了许多疾病的遗传基础。不幸的是，相当可观的与购买和运行专门设备有关的费用和对专用实验室基础设施的要求，配备了高度目前，训练有素的人员使排序在日常工作中使用不切实际医疗实践。微组装测序设备准备提供一种传统设备的廉价替代品，最终使直接基因组测序可用于真正的医疗诊断协议。此外，转移到微设备格式大大减少了分析所需的样品体积(纳升数量级)和介绍制造自含式便携式测序设备的能力它可以轻松地部署在偏远的现场位置，用于各种应用，包括快速识别和跟踪传染性疾病疾病。任何测序设备的一个关键要素是能够根据大小执行测序用电泳法分离DNA片段。完全测序需要足够的分辨率才能区分仅有差异的片段长度接近1,000个碱基对的片段中的一个碱基对长度-这一要求通常由传统的板材凝胶和毛细管电泳法。目前，分离通道要实现单碱基，需要几厘米量级的长度微细加工电泳系统中的配对拆分。尽管令人印象深刻，当前一代的微型测序设备的尺寸仍然太大大量利用可能实现的巨大成本节约的一小部分通过光刻制造技术进行大规模生产。这些半导体行业中经常使用的技术，允许数十或在一片晶圆上生产数百个器件。这样做的具体目的是研究项目是证明基于微设备的电泳法分离可以达到与传统大尺度DNA相当的分辨率分析系统的长度在一厘米或更小的数量级上。这个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而分子生物学方法则是为了满足这一研究目标。