Real-time DNA Sequencing

实时 DNA 测序

• 批准号: 7242588
• 负责人: Susan H Hardin
• 金额: $ 126.75万
• 依托单位: VISIGEN BIOTECHNOLOGIES, INC.
• 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 2008-08-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7242588
• 关键词: Real; time; DNA; Sequencing

DESCRIPTION (provided by applicant): VisiGen Biotechnologies, Inc. is developing a sequencing system in which polymerase and nucleotides act together as direct molecular sensors of DNA base identity. More specifically, the technology detects the interaction between a modified nucleotide and a fluorescently-modified polymerase. As a nucleotide is incorporated into the growing DNA polymer, energy transfers from an excited donor fluorophore within the polymerase to an acceptor fluorophore within the nucleotide, stimulating the emission of a base-specific incorporation signature that is directly detected in real-time. Cutting-edge technologies, including single molecule detection, fluorescent molecule chemistry, computational biochemistry, and biomolecule engineering, are combined to create this novel sequencing system. DNA samples will be processed in massively parallel arrays that will enable large genomes to be sequenced in less than a day for approximately $1000 and permit sufficient oversampling to produce redundant data that will minimize errors. The system is being developed to identify pathogens (or variations thereof) and to enable comprehensive genome analysis. Proof of principle for VisiGen's technology is demonstrated: sequence information has been detected at the single molecule level in real-time using an immobilized, donor-labeled polymerase and 2 different acceptor-labeled dNTPs. Given the success of our federally-funded efforts, further development of the VisiGen system to sequence arrays of single DNA molecules in real-time is clearly warranted. The specific aims of the proposed research are to refine the sequencing chemistry and to design, build and test the next generation single-molecule DNA sequencing instrument. This instrument, Alpha-1, will be placed at VisiGen and accelerate technology development. This instrument will be used for beta testing by researchers at the Baylor College of Medicine, Human Genome Sequencing Center (Houston, TX) and SeqWright, Inc. (Houston, TX). Feedback from personnel involved in these beta tests will be incorporated into the next phase of technology development.

描述（由申请人提供）：Visigen Biotechnologies，Inc。正在开发一个测序系统，其中聚合酶和核苷酸将作为DNA碱基认同的直接分子传感器一起起作用。更具体地说，该技术检测到修饰的核苷酸与荧光改性的聚合酶之间的相互作用。由于将核苷酸掺入了生长的DNA聚合物中，因此能量从聚合酶内的激发供体荧光团转移到核苷酸内的受体荧光团，刺激了直接检测到实时检测的基础特异性掺入签名的发射。尖端技术，包括单分子检测，荧光分子化学，计算生物化学和生物分子工程，以创建这种新型的测序系统。 DNA样品将以大规模平行的阵列进行处理，这将使大型基因组在不到一天的时间内以约1000美元的价格进行测序，并允许足够的过度采样以产生冗余数据，以最大程度地减少错误。正在开发该系统以识别病原体（或其变异）并实现全面的基因组分析。 证明了Visigen技术的原理证明：使用固定的供体标记的聚合酶和2个不同的受体标记的DNTP在单个分子水平上在单分子水平上检测到了序列信息。鉴于我们由联邦资助的努力取得了成功，因此，明确保证了尼古包系统的进一步发展为实时的单个DNA分子阵列的序列。拟议研究的具体目的是完善测序化学，并设计，构建和测试下一代单分子DNA测序仪器。该仪器Alpha-1将放置在Visigen并加速技术开发。该乐器将用于贝勒医学院，人类基因组测序中心（德克萨斯州休斯顿）和Seqwright，Inc。（德克萨斯州休斯顿）的研究人员的Beta测试。参与这些Beta测试的人员的反馈将纳入下一阶段的技术开发阶段。