Electronic Sequencing in Nanopores
纳米孔中的电子测序
基本信息
- 批准号:7238487
- 负责人:
- 金额:$ 168.58万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2005
- 资助国家:美国
- 起止时间:2005-08-01 至 2008-08-18
- 项目状态:已结题
- 来源:
- 关键词:AdsorptionAgreementAgricultureAlgorithmsBase CompositionBiophysicsCarbon NanotubesCellsCharacteristicsCollaborationsComputer-Assisted Image AnalysisCopy Number PolymorphismDNADNA SequenceDetectionDevelopmentDevicesDiagnosisDiseaseEcologyElectrodesElectronicsEngineeringEvolutionFutureGenomeGenomicsGoalsHealthHourHumanHuman GeneticsIndividualIonsJointsLabelLengthLicensingLinkLocalizedMeasurementMechanicsMembraneMethodsMicrofluidicsMolecularNanotubesNeckNoiseNucleic AcidsNucleotidesOrangesPerformancePhasePhysicsPolymersProcessProductionPropertyQuality ControlRNARateReadingResearchResolutionRightsRiskSchemeScienceServicesSideSignal TransductionSingle-Stranded DNASolutionsSpeedSurfaceSystemTechnologyTemperatureTestingTimeVariantWorkakesonbasecomputerized data processingcostdata acquisitiondetectordisorder preventionelectrical propertyimprovedinstrumentinterestmammalian genomemonomernanoporenanoscalequantumremediationresearch studyresponsescale upsensorsingle moleculesolid statetheoriesvoltage
项目摘要
DESCRIPTION (provided by applicant): The long-term objective is to develop a general utility instrument capable of inexpensive de novo sequencing that can also be used for re-sequencing projects to recognize genome variation in heterozygous genomes. The system being developed will sequentially, and directly, identify the nucleotides in very long fragments of genomic DNA from a base-dependent electronic signal produced by a nanopore articulated with nanotube probes. 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 and minimal preparative steps. The specific aims for the initial 5 year period of this project are: 1. Improve nanopore surfaces to reduce nonspecific adsorption, pore clogging, and electrical noise; 2. Fabricate and test a nanopore detector articulated with integrated nanotubes for molecular identification; 3. Investigate and optimize the electronic properties of nanotube-DNA interactions to control DNA translocation, orientation and nucleotide contrast; 4. Develop new enzymatic methods to better control and limit the rate of DNA translocation through articulated nanopores; 5. Develop algorithms for feature detection and identification of signals from articulated nanopores; 6. Demonstrate single base sensitivity and resolution on single-stranded DNA translocating through a nanopore. If, as proposed here, we are able to resolve each base as it passes through a nanopore at the rate of 104 bases/sec, an instrument with an array of 100 such nanopores could produce a high-quality draft sequence of 1 mammalian genome in ~20 hours at a cost of approximately $1,000/mammalian genome. Genomic sequencing at these sharply reduced costs would make vital contributions to improved human health on many fronts, including the understanding, diagnosis, treatment, and prevention of disease; advances in agriculture, environmental science and remediation; and the genetics of human health and disease derived from the understanding of evolution.
描述(由申请人提供):长期目标是开发一种通用仪器,能够进行廉价的从头测序,也可用于重新测序项目以识别杂合基因组中的基因组变异。正在开发的系统将根据由纳米管探针连接的纳米孔产生的碱基依赖性电子信号,连续、直接地识别非常长的基因组 DNA 片段中的核苷酸。最终系统旨在使用来自少于 100 万个细胞的 DNA 提供 6.5 倍基因组覆盖率的相对高质量序列,无需扩增且制备步骤最少。该项目最初 5 年的具体目标是: 1. 改善纳米孔表面,减少非特异性吸附、孔堵塞和电噪声; 2. 制作并测试与集成纳米管连接的纳米孔探测器,用于分子识别; 3. 研究并优化纳米管-DNA相互作用的电子特性,以控制DNA易位、方向和核苷酸对比; 4. 开发新的酶促方法,以更好地控制和限制DNA通过铰接纳米孔的易位速率; 5. 开发铰接纳米孔信号的特征检测和识别算法; 6. 展示单链 DNA 通过纳米孔易位的单碱基灵敏度和分辨率。如果按照此处提出的那样,我们能够在每个碱基以 104 个碱基/秒的速率通过纳米孔时对其进行解析,则具有 100 个此类纳米孔阵列的仪器可以在约 20 小时,每个哺乳动物基因组的成本约为 1,000 美元。成本大幅降低的基因组测序将为在许多方面改善人类健康做出重要贡献,包括疾病的理解、诊断、治疗和预防;农业、环境科学和修复方面的进步;人类健康和疾病的遗传学源自对进化的理解。
项目成果
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