MICROFABRICATION OF AN INTEGRATED DNA SEQUENCING SYSTEM

临床和临床对DNA序列信息的预期需求 基础研究实验室，几乎是无限的。生化和生物化学 DNA测序的分析操作有很好的特点，但 还没有组装成一个简单的集成系统。基于 既定的方法，这项建议将发展样品处理， 用于完全集成的电泳、检测和控制组件 DNA测序技术。完整的集成确保每个样本 进入该系统将拥有一套独特的专用设备， 从而消除了样品处理的瓶颈。这些组件将是 利用光刻微细加工技术在晶体上形成 硅胶或玻璃。 光刻制造允许将组件设计为 兼容性，易于组装成新颖的组合，并且价格低廉 制作。先进的硅和玻璃薄膜制造方法可以 制造各种机械和电子设备--从 毫米到亚微米大小。由于硅的特性- 基于材料的很好地理解，设计可以快速产生， 使用计算机辅助设计软件进行测试和复制。 拟议的工作将开发一个集成的DNA测序系统， 包括样品处理和电泳法分离，仅使用 硅或玻璃光刻技术。系统(I)将 需要操作员与样品液体进行最小程度的交互，(Ii)将 提供对生化反应的广泛实时控制，(Iii)将 从测序中直接生成电子输出数据作为波段 电泳胶，以及(Iv)将能够结合反馈 来自样本的信息，供下游决策点使用。这个 系统将对纳升体积的液体样本进行操作，并包括 用于控制样品位置、温度、光电检测和 电泳法。 该项目有三个具体目标：(1)建造微型高地 分辨率电泳仪，(2)液电一体化 和(3)取消排序 使用智能系统的瓶颈。前两个目标将 论证建造DNA测序设备的基本可行性 来自固态材料。目标3将结合特定于序列的 具有一般液体样品处理部件的部件，以生产 全功能设备。

The anticipated demand for DNA sequence information, in both clinical and basic research laboratories, is virtually unlimited. The biochemical and analytical manipulations for DNA sequencing are well-characterized but have not been assembled into a simple integrated system. Based on established methods, this proposal will develop the sample handling, electrophoresis, detection, and control components for a fully integrated DNA sequencing technology. Complete integration ensures that each sample entering the system will have a unique, dedicated set of equipment, thereby eliminating sample processing bottlenecks. The components will be formed using photolithographic microfabrication techniques on crystalline silicon or glass. Photolithographic fabrication allows components to be designed for compatibility, easy assembly into novel combinations, and inexpensive mass production. Advanced silicon and glass thin-film fabrication methods can make a wide range of mechanical and electronic devices -- ranging from millimeter to submicron in size. Since the characteristics of silicon- based materials are well understood, designs can be rapidly generated, tested, and replicated using computer-aided design software. The proposed work will develop an integrated DNA sequencing system, including sample handling and electrophoretic separation, using only silicon or glass photolithographic techniques. The system (i) will require minimal operator interaction with the sample liquids, (ii) will provide extensive real-time control of biochemical reactions, (iii) will directly generate electronic output data as bands from sequencing electrophoresis gels, and (iv) will be able to incorporate feedback information from samples for use at downstream decision points. The system will operate on nanoliter volumes of liquid samples and include circuitry for control of sample location, temperature, photodetection, and electrophoresis. The project has three specific aims: (1) construction of a miniature high resolution electrophoresis system, (2) fluidic and electronic integration of the sequencing system, and (3) the elimination of sequencing bottlenecks using intelligent systems. The first two aims will demonstrate the basic feasibility of constructing DNA sequencing equipment from solid-state materials. Aim 3 will combine the sequencing-specific components with general liquid sample handling components to produce a fully functional device.