PFI:AIR - TT: Dielectrophoretic Fractionation of DNA for Next Generation Sequencing

PFI:AIR - TT：用于下一代测序的 DNA 介电泳分离

• 批准号: 1445006
• 负责人: Alexandra Ros
• 金额: $ 20万
• 依托单位: Arizona State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-15 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1445006&HistoricalAwards=false
• 关键词: PFI; AIR; TT; Dielectrophoretic; Fractionation

This PFI:AIR Technology Translation project focuses on nucleic acid fractionation to fill the need for adequate sample preparation for the rapidly increasing field of next generation genome sequencing. The microfluidic nucleic acid sorter is important because genome sequencing is fundamental for deciphering biological pathways leading to malfunction and disease. All sequencing technologies require the fractionation of nucleic acids tailored to the specific sequencing approach. This project will result in a proof-of-concept design for a microfluidic nucleic acid sorter. This microfluidic nucleic acid sorter has the following unique features: a dielectrophoretic process fractionating nucleic acids by size and conformation, realization on a microfluidic platform and operation under continuous flow. These features provide the advantages of nucleic acid fractionation in tunable size ranges, cost effectiveness, minute sample amounts and applicability to both DNA and RNA when compared to the leading competing shearing technology and bead purification technology in this market space. This project addresses previously little explored technology gaps in next generation sequencing as it translates from research discovery toward commercial application, comprised of the development of a versatile device applicable to a wide range of nucleic acid lengths, conformations and nucleic acid types and, the micro-preparative approach realized through microfluidic integration. Specifically, the project aims in the development of nucleic acid array and constriction fractionators based on insulator-based dielectrophoresis suitable for fragment lengths between 100 bp and ~20 kbp and in volumes and quantities applicable for next generation sequencing. The design of the dielectrophoretic fractionation process will be optimized with the aid of numerical simulation tools and improved device fabrication technologies. A systematic study of size fractionation capabilities of the proof-of-principle devices for DNA and RNA will be performed and optimized fractions will be tested on existing next generation sequencing platforms. The versatility of the approach will allow the application for existing but also future next generation sequencing platforms and their requirements in nucleic acid sample preparation.In addition, personnel involved in this project, undergraduate students and post docs, will receive innovation and technology translation experiences through active involvement in the research components of this project, the proof-of-principle development and the collaboration with an industry partner. Mentoring activities further fosters the participation of women in the research aspects of this project.

这个PFI：AIR技术翻译项目专注于核酸分离，以满足快速增长的下一代基因组测序领域对充分样品制备的需求。微流控核酸分选仪很重要，因为基因组测序是破译导致故障和疾病的生物途径的基础。所有测序技术都需要根据特定的测序方法对核酸进行分级分离。该项目将导致微流体核酸分选机的概念验证设计。这种微流控核酸分选仪具有以下独特的特点：介电泳过程通过大小和构象分级核酸，在微流控平台上实现和在连续流动下操作。与该市场领域领先的竞争性剪切技术和珠纯化技术相比，这些特征提供了核酸分级的优势，包括可调尺寸范围、成本效益、微量样品量以及对DNA和RNA的适用性。该项目解决了以前很少探索的下一代测序技术差距，因为它从研究发现转化为商业应用，包括开发适用于各种核酸长度，构象和核酸类型的通用设备，以及通过微流体集成实现的微制备方法。具体而言，该项目旨在开发基于绝缘体介电电泳的核酸阵列和收缩分馏器，适用于100 bp至~20 kbp的片段长度，并适用于下一代测序的体积和数量。介电电泳分馏过程的设计将在数值模拟工具和改进的器件制造技术的帮助下进行优化。将对DNA和RNA的原理验证装置的大小分级能力进行系统研究，并将在现有的下一代测序平台上测试优化的级分。该方法的多功能性将允许应用于现有的以及未来的下一代测序平台及其在核酸样品制备中的要求。此外，参与该项目的人员，本科生和博士后，将通过积极参与该项目的研究组成部分获得创新和技术翻译经验，原理验证开发以及与行业合作伙伴的合作。指导活动进一步促进妇女参与该项目的研究工作。