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

允许成本有效和准确的从头测序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所需的见解。成本 仍然太高，质量不足，使测序技术负担得起的常规使用， 基因组学在个人健康护理中的应用“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 相对于现有技术系统的吞吐量和易用性。 总体目标是开发一个使用纳米磁电子平台的测序系统。我们 提出了将样品制备和富集步骤与测序模块集成的系统。 样品制备将依赖于一个可重复使用的磁电子芯片和一个新的无乳液放大 法扩增后，通过“分选”实现携带单克隆珠的模板的富集。 基于DNA电荷然后将珠子转移并保持在与微磁体阵列集成的微磁体阵列上。 由基于CMOS的纳米电子传感器的高密度阵列组成的测序传感器芯片， 检测延伸反应。代替反相乳液，我们正在开发一种易于使用的芯片型 该方法将联合收割机多个样品处理步骤组合在单个装置中。“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。有效 珠粒的捕获、DNA和扩增产物的浓缩和限制，可以允许 消除了全基因组扩增及其固有的偏差。可重复使用的“虚拟纳米反应器”芯片 在整个芯片上提供均匀的反应条件。该装置将消除反应的可变性 体积和双泊松分布固有的乳液PCR。可重复使用的“无孔”测序 传感器芯片提供高灵敏度的检测，高效和均匀的试剂输送和清洗， 并通过最小化去相位允许更长的读取和更高的准确性 并提供反应的均匀性，还减少了试剂消耗和成本。的可重用性 芯片显著降低了耗材成本。总之，简化的自动化工作流程， 试剂消耗，使用未修饰的核苷酸和聚合酶的无标记电子检测导致 显著节省成本和提高精度。 我们的目标是开发一个平台，以约50美元的消耗成本对基因组进行测序， 高达1000个基座，预装配精度> 99.7%， 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.