Fabrication of a dual nano-biosensor for accurate sensitive and inexpensive elec

下一代测序(NGS)的进展极大地促进了基础生命科学研究 但需要在技术上取得更多突破，才能在临床环境中常规使用测序 用于个性化医疗，作为疾病诊断、预后和治疗工具等应用。 今天的仪器和检测平台允许对整个基因组进行测序，成本为 大约10,000美元，需要对昂贵的设备进行大量前期投资。当前的瓶颈 包括使用昂贵试剂和光学检测系统、大量数据存储和处理 要求、样品制备速度慢和周转时间长。因此，总体目标是发展一个低 成本、实时、完全集成的排序系统，涵盖整个工作流程，包括 几个正在并行开发的创新的样品制备和测序模块。 测序模块的核心组件之一将是两个新型的电动纳米生物传感器，它们能够 无标记、准确、灵敏的pH变化检测是聚合反应的电信号 合成测序(SBS)过程中的反应。与电子测序相比，电子测序有几个优势 焦磷酸测序和其他现有技术，因为它不需要荧光标记或报告酶 和光学探测系统。两个独立运行的电子纳米传感器的创新使用将 以更低的成本生成更准确的测序数据，同时减少对 冗余序列覆盖。 同时检测稳态信号(如纳米针)和暂态信号的双重传感机制 (纳米桥)提供更高级别的置信度并提高基本调用精度，因为两个正交 两个传感器的感应机制报告了相同的事件。这项提案的总体目标是 证明了使用纳米桥进行电子DNA测序的双重传感的可行性 纳米针生物传感器和开发双传感器检测模块的设计。基本功能测试 在比较由pH变化引起的信号产生之后，将确定对pH的响应 在‘合成测序’聚合反应中。在第一阶段，我们将致力于 低阈值耗尽型纳米电阻纳米桥生物传感器阵列的表征 大的线性检测范围及两种纳米传感器的性能比较 对pH或电荷的响应、缓冲离子强度和DNA测序方面的性能。结果是 为今后双传感器阵列的设计和建模提供了依据。

Advances in Next Generation Sequencing (NGS) have greatly accelerated fundamental life science research but additional breakthroughs in technology are required to allow routine use of sequencing in the clinical setting for personalized medicine as a disease diagnostic, prognostic and therapeutic tool and other applications. Today's instrumentation and detection platforms allow sequencing of an entire genome for a cost of approximately $10,000 requiring large up-front investment into expensive equipment. The current bottlenecks include the use of expensive reagents and optical detection systems, large data storage and processing requirements, slow sample preparation and turnaround times. Therefore, the overall goal is to develop a low cost, real-time, fully-integrated sequencing system that encompasses the entire workflow and consists of several innovative sample preparation and sequencing modules that are being developed in parallel. One of the core components of the sequencing module will be two novel electric nano-biosensors that enable label free, accurate and sensitive detection of the pH change as an 'electrical signature' of the polymerization reaction during 'sequencing by synthesis' (SBS). Electronic sequencing offers several advantages over pyrosequencing and other existing techniques as it does not require fluorescent labels or reporter enzymes and optical detection systems. The innovative use of two independently operating electric nanosensors will generate significantly more accurate sequencing data at lower cost, while reducing the need for highly redundant sequence coverage. Dual sensing mechanism of detecting both steady state signal (e.g. nanoneedle) and transient signal (nanobridge) provides higher level of confidence and improves base calling accuracy, as two orthogonal sensing mechanisms with the two sensors are reporting the same event. The overall goal for this proposal is to demonstrate the feasibility of dual sensing for electronic DNA sequencing using the nanobridge and nanoneedle biosensors and developing a design for the dual sensor detection module. Basic functional testing and response to pH will be determined followed by a comparison of signal generation induced by pH changes during 'sequencing by synthesis' polymerization reaction. In the first phase, we will work on the characterization of nanobridge biosensor array, which is a depletion mode nano-resistor with low threshold and large linear detection range and the comparison of the properties of the two nanosensors with respect to response to pH or charge, buffer ionic strength and performance for DNA sequencing perspective. The results from the feasibility study will provide the basis for the design and modeling of a dual sensor array in the future.