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.

该PFI：空气技术翻译项目的重点是核酸分馏，以满足对下一代基因组测序迅速增加领域的适当样品制备的需求。微流体核酸分散剂很重要，因为基因组测序对于破门的生物学途径是基础，导致故障和疾病。所有测序技术都需要根据特定测序方法量身定制的核酸分馏。该项目将为微流体核酸分散器提供概念验证设计。该微流体核酸分散器具有以下独特特征：按大小和构象分级的介电摄取过程分级分级，在微流体平台上实现以及在连续流下的操作。与领先的竞争剪切技术和该市场空间中领先的竞争剪切技术和珠子纯化技术相比，这些功能提供了可调节尺寸范围，成本效益，样本量以及对DNA和RNA的适用性的优势。该项目以前几乎没有探索下一代测序中的技术差距，因为它从研究发现转化为商业应用，包括用于开发适用于广泛的核酸长度，构象和核酸类型的多功能设备，以及通过微富集整合实现的微预览方法。具体而言，该项目的目的是基于基于绝缘体的介电的核酸阵列和收缩分级器的发展，适用于适用于100 bp至〜20 kbp之间的片段长度以及适用于下一代测序的体积和数量。介电性分级过程的设计将借助数值模拟工具和改进的设备制造技术来优化。将对现有的下一代测序平台进行测试，将对DNA和RNA的原理设备的尺寸分馏能力进行系统研究。该方法的多功能性将允许应用于现有但未来的下一代测序平台及其在核酸样品准备中的要求。此外，参与该项目的人员，本科生和文档的人员将通过积极参与该项目的研究组件，并与该项目的研究组成部分获得创新和技术翻译体验。指导活动进一步促进了妇女参与该项目的研究方面。