Electronic Sequencing in Nanopores

描述（由申请人提供）： 长期目标是一种纳米孔检测器芯片，用于能够进行廉价从头测序的通用仪器，也可用于重新测序项目。该仪器直接产生碱基依赖性电子信号，因为单链基因组DNA的多酶长度片段被顺序驱动通过与电接触的单壁碳纳米管探针铰接的纳米孔。最终的系统旨在使用来自少于100万个细胞的DNA，在不扩增或标记的情况下，提供相对高质量的基因组覆盖率≥ 6.5倍的序列。具体目标是：1）表征在具有和不具有DNA分子的离子溶液中的无缺口纳米管铰接纳米孔检测器以建立装置模型; 2）控制ssDNA在暴露于无缺口纳米管铰接纳米孔中的纳米管表面上的结合、移位和滑动; 3）研究和优化DNA分子诱导的场效应对无缺口纳米管铰接纳米孔中纳米管电极电导的调制作为纳米管偏压的函数，门电压和溶液性质; 4）分析和优化第一代检测器中有间隙的纳米管电极之间的隧穿电流调制; 5）设计和制造具有嵌入的“T”纳米管几何形状的第二代检测器，并在Kb长度的DNA链上实现1 Kb/sec测序; 6）设计用于高通量10 Kb/sec/纳米孔测序的第三代纳米孔检测器。如果我们能够像这里提出的那样以104个碱基/秒的速率解析每个碱基通过纳米孔，则具有100个这样的纳米孔的阵列的仪器可以在约20小时内以大约1，000美元/哺乳动物基因组的成本产生一个哺乳动物基因组的高质量草图序列。基因组测序在这些降低成本将作出重要贡献，改善人类健康在许多方面，包括理解，诊断，治疗和预防疾病;环境科学和补救;和人类健康和疾病的遗传学从进化的理解。 我们正在开发一种仪器的核心检测器，该仪器可以在大约20小时内以大约1,000美元/哺乳动物基因组的成本产生一个哺乳动物基因组的高质量草图序列。基因组测序在这些降低成本将作出重要贡献，改善人类健康在许多方面，包括理解，诊断，治疗和预防疾病;环境科学和补救;和人类健康和疾病的遗传学从进化的理解。

DESCRIPTION (provided by applicant): The long-term objective is a nanopore detector chip for a general utility instrument capable of inexpensive de novo sequencing that can also be used for re-sequencing projects. The instrument directly generates base-dependent electronic signals as multi-kilobase length fragments of single stranded genomic DNA is driven sequentially through nanopores articulated with electrically contacted single walled carbon 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 or labeling. The specific aims are: 1) Characterize ungapped nanotube articulated nanopore detectors in ionic solution with and without DNA molecules to establish a device model; 2) Control ssDNA binding, translocation, and sliding on the nanotube surface exposed in ungapped nanotube articulated nanopores; 3) Study and optimize DNA molecule induced field effect modulation of nanotube electrode conductance in ungapped nanotube articulated nanopores as a function of nanotube bias, gate voltage, and solution properties; 4) Analyze and optimize tunneling current modulations between gapped nanotube electrodes in the first generation detector; 5) Design and fabricate a second generation detector with embedded 'T' nanotube geometry and achieve 1 Kb/sec sequencing on Kb length strands of DNA; 6) Design a third generation nanopore detector for high throughput 10 Kb/sec/nanopore sequencing. If we are able to resolve each base as it passes through a nanopore at the rate of 104 bases/sec as proposed here, an instrument with an array of 100 such nanopores could produce a high quality draft sequence of one mammalian genome in ~20 hours at a cost of approximately $1,000/mammalian genome. Genomic sequencing at these reduced costs would make vital contributions to improved human health on many fronts, including the understanding, diagnosis, treatment, and prevention of disease; environmental science and remediation; and the genetics of human health and disease derived from the understanding of evolution. PROJECT HEALTH RELEVANCE We are developing the core detector of an instrument that could produce a high-quality draft sequence of one mammalian genome in ~20 hours at a cost of approximately $1,000/mammalian genome. Genomic sequencing at these reduced costs would make vital contributions to improved human health on many fronts, including the understanding, diagnosis, treatment, and prevention of disease; environmental science and remediation; and the genetics of human health and disease derived from the understanding of evolution.