Electronic Sequencing in Graphene Nanopores

石墨烯纳米孔中的电子测序

• 批准号: 8359840
• 负责人: DANIEL none BRANTON
• 金额: $ 120万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8359840
• 关键词: Agriculture; Cells; DNA; Devices; Diagnosis; Discrimination; Ecology; Ecosystem; Electronics; Elements; Evolution; Future; Genome; Genomics; Growth; Hour; Human; Human Genome; Label; Life; Lipid Bilayers; Membrane; Methods; Nucleotides; Polymers; Pore Proteins; Procedures; Process; Protein Array; Research; Resolution; Societies; System; Teflon; base; cost; detector; disorder prevention; improved; instrument; mammalian genome; nanopore; particle beam; remediation; tool; tool development

Our research has shown that a single layer of graphene is an ideal membrane in which to fabricate high resolution nanopore detectors to sense the presence of single DNA molecules and their nucleobases. Our objective is to develop the tools and procedures needed to realize a scalable nanopore sequencing device which will significantly reduce future de novo sequencing cost by directly identifying the nucleobases on single stranded genomic DNA molecules that are driven sequentially through an array of precisely dimensioned graphene nanopores. The final system is intended to provide a relatively high quality sequence from >6.5-fold coverage of a genome using DNA from fewer than 1 million cells, with no amplification or labeling. The specific aims are to: a) implement a graphene edge-sputtering process to facilitate high precision fabrication of nanopore arrays; b) optimize discrimination between the four nucleotides of DNA using ionic current blockades or in-plane conductivity change when ssDNA polymers are driven through graphene nanopores; and c) support lipid bilayers across graphene apertures to enhance the feasibility of parallel recordings from arrays of protein pores. Successful completion of these aims will provide the key building blocks of a nanopore sequencing device that can accurately sequence an entire human genome at a cost of less than $1,000. The ability to inexpensively and accurately sequence complete genomes has the potential of remarkably improving many facets of human life and society, including the understanding, diagnosis, treatment and prevention of disease.

我们的研究表明，单层石墨烯是一种理想的膜，可以用来制造高分辨率的纳米孔检测器，以检测单个DNA分子及其核碱基的存在。我们的目标是开发实现可扩展的纳米孔测序装置所需的工具和程序，这将显着减少未来的从头测序 通过直接识别单链基因组DNA分子上的核碱基，这些DNA分子被顺序驱动通过精确尺寸的石墨烯纳米孔阵列。最终的系统旨在使用来自少于1百万个细胞的DNA，在没有扩增或标记的情况下，从>6.5倍的基因组覆盖度提供相对高质量的序列。具体 目标是：a）实施石墨烯边缘溅射工艺，以促进纳米孔阵列的高精度制造; B）当ssDNA聚合物被驱动通过石墨烯纳米孔时，使用离子电流阻断或面内电导率变化来优化DNA的四种核苷酸之间的区分;以及c）支撑脂质双层穿过石墨烯孔，以 增强蛋白质孔阵列平行记录的可行性。这些目标的成功完成将为纳米孔测序设备提供关键的构建模块，该设备可以以低于1000美元的成本准确地对整个人类基因组进行测序。廉价而准确地对完整基因组进行测序的能力有可能显著改善人类生活和社会的许多方面，包括对疾病的理解、诊断、治疗和预防。