INTEGRATED DNA SEQUENCING SYSTEM

描述：（申请人的摘要）人类基因组计划正在迅速地 开发人类基因组的共识序列草案。的巨大价值 这一信息引发了生产率的规模扩大， 结合高通量毛细管阵列电泳仪器。 需要进一步改进以降低样品成本，提高自动化程度， 并提高吞吐率。 该项目将创建一个自动化的集成DNA分析系统， 制备纳升（nL）规模的测序反应，并分析反应 在一个超高通量的，基于微芯片的DNA测序仪上。集成 系统将消耗今天使用的试剂的百分之一左右， 十倍的高吞吐率。综合系统将包括两个主要部分 模块、样品制备模块和超高通量DNA分析 module.样品制备模块将使用384个毛细管阵列， 用于移动液体并进行125 nL体积的生化反应的试剂盒。 该模块将从裂解的克隆开始自动进行鸟枪测序， 产生荧光标记的循环测序反应，用于毛细管或 微芯片分析基于微芯片的DNA分析模块将加载，运行， 在微通道中检测DNA样品，用于测序和基因分型。本模块 将使用机器人将样品从微量滴定板转移到样品加载 微型芯片上的端口，将微型芯片与探测器下的电极对接， 注入样品并分离它们。将开发这两个模块 同时进行，然后集成到完整的自动测序系统中。 该项目将开发一系列原型，这些原型将在2015年进行alpha测试。 分子动力学生产测序组，然后在我们的 华盛顿大学联合基因组研究所的合作者 测序中心和大学的大分子分析设施 威斯康星-麦迪逊外部测试的结果将用于 验证系统性能，完善原型，并简化 将这项新技术整合到基因组中心和核心设施中。

DESCRIPTION: (Applicant's Abstract) The Human Genome Project is rapidly developing a draft consensus sequence of the Human Genome. The immense value of this information has triggered the scale up of production rates and the incorporation of high-throughput capillary-array electrophoresis instruments. Further improvements are needed to decrease sample costs, increase automation, and improve throughput rates. This project will create an automated, integrated DNA analysis system that prepares nanoliter (nL)-scale sequencing reactions and analyzes the reactions in an ultrahigh-throughput, microchip-based DNA sequencer. The integrated system will consume about one-hundredth of the reagents used today and have a ten-fold higher throughput rate. The integrated system will consist of two main modules, a sample preparation module and an ultrahigh-throughput DNA analysis module. The sample preparation module will use arrays of 384 capillaries in \cassettes to move liquids and perform biochemical reactions in 125 nL volumes. This module will automate shotgun sequencing beginning with lysed clones to produce fluorescently labeled cycle sequencing reactions for capillary or microchip analysis. The microchip-based DNA analysis module will load, run, and detect DNA samples in microchannels for sequencing and genotyping. This module will use robotics to transfer samples from microtiter plates to sample loading ports on a microchip, dock the microchip with electrodes under a detector, inject samples and separate them. The two modules will be developed simultaneously and then integrated into a complete automated sequencing system. The project will develop a series of prototypes that will be alpha-tested in the Molecular Dynamics Production Sequencing Group and then beta-tested at our collaborators at the Joint Genome Institute, the Washington University Genome Sequencing Center, and the Macromolecular Analysis Facility at the University of Wisconsin-Madison. The results of the external testing will be used to verify the system performance, refine the prototypes, and streamline the integration of this new technology into genome centers and core facilities.