Low Cost Sequencing with Re-usable Magnetic Arrays and Nanoelectronic Sensors

能够实现经济高效和准确的从头开始和重新测序基因组的测序方法对于 为个人层面的人类健康、疾病管理和诊断提供必要的见解。费用 仍然太高，质量不够高，无法使测序技术负担得起常规使用 基因组学在个人保健中的应用。“基因电子纳米集成超灵敏”(GENIUS)平台 基于创新技术，在成本、准确性、读取长度、 与当前最先进的系统相比，吞吐量和易用性更好。 总体目标是开发一种使用纳米磁电平台的测序系统。我们是 提出了一种将样品制备和浓缩步骤与测序模块相结合的系统。 样品的制备将依赖于可重复使用的磁电子芯片和一种新型的无乳胶放大 方法。扩增后，通过‘分选’实现了携带单克隆珠的模板的富集化 根据DNA指控。然后将珠子转移并保持在集成了 由高密度的基于cmos的纳米电子传感器阵列组成的序列传感器芯片，用于直接测量 延伸反应的检测。我们正在开发一种易于使用的、基于芯片的反相乳剂，以取代反相乳剂 将在单个设备中结合多个样品处理步骤的方法。“虚拟”的目的 纳米反应器芯片是在高密度的单珠阵列上产生克隆扩增模板。高效的 捕获珠子，浓缩和限制DNA和扩增产物，可以允许 消除全基因组扩增及其固有的偏见。可重复使用的虚拟纳米反应堆芯片 在整个芯片上提供统一的反应条件。该装置将消除反应中的可变性 乳胶聚合酶链式反应固有的体积和双泊松分布。可重复使用的“无井”测序 传感器芯片提供高灵敏度的检测，高效而均匀的试剂输送和洗涤相结合 具有高效率的磁珠捕获，并通过最大限度地减少去相来允许更长的读取时间和更高的精度 并且提供了反应的一致性，还降低了试剂消耗和成本。的可重用性 芯片显著降低了耗材成本。总而言之，简化的自动化工作流具有低成本 试剂消耗，使用未经修饰的核苷酸和聚合酶的无标记电子检测导致 显著节省成本并提高准确性。 我们的目标是开发一个对基因组进行测序的平台，耗材成本约为50美元，平均读取长度 最多1000个底座，预装精度为99.7%，成本降低和简化程度与 样品制备。

Sequencing methods allowing cost effective and accurate de novo and re-sequencing genomes are critical to provide needed insights for human health, disease management and diagnostics at the individual level. Costs still remain too high with inadequate quality, to make sequencing technologies affordable for the routine use of genomics in individual health care. The "Gene Electronic Nano-Integrated Ultra-Sensitive" (GENIUS) platform is based on innovative technologies to provide significant improvements in cost, accuracy, read length, throughput and ease of use relative to current state-of-art systems. The overall goal is to develop a sequencing system using nano-magnetic-electronic platforms. We are proposing a system that integrates sample preparation and enrichment steps with the sequencing module. Sample preparation will rely on a re-usable magnetic-electronic chip and a novel emulsion-free amplification method. After amplification, enrichment of template carrying monoclonal beads is achieved through 'sorting' based on DNA charge. Beads are then transferred and held on a micromagnet array integrated with a sequencing sensor chip consisting of high-density arrays of CMOS-based nano-electronic sensors for direct detection of extension reactions. In lieu of a reverse emulsion we are developing an easy to use, chip-based approach that will combine multiple sample processing steps in a single device. The purpose of the 'Virtual Nano-Reactor' chip is to generate clonal amplified template on high-density array of single beads. The efficient capture of beads, concentration and confinement of DNA and amplification products, may permit the elimination of whole genome amplification with its inherent bias. The re-usable 'Virtual Nano-reactor' chip provides uniform reaction conditions across the entire chip. The device will eliminate the variability in reaction volumes and double-Poisson distribution inherent in emulsion PCR. The re-usable "well-free" sequencing sensor chip provides high-sensitivity detection, efficient and uniform reagent delivery and washing, combined with the high efficiency bead capture and permits longer reads and higher accuracy by minimizing de-phasing and providing uniformity of reaction, and also reduce reagent consumption and cost. The re-usability of the chip significantly reduces the consumable cost. In summary, the simplified automatable workflow with low reagent consumption, label-free electronic detection using unmodified nucleotides and polymerase results in significant cost savings and improved accuracy. Our goal is to develop a platform to sequence a genome with consumable costs of ~$50, average read length of up to 1000 bases, and pre-assembly accuracy of > 99.7% with similar cost reduction and simplification for sample preparation.