DEVELOPMENT OF A NEW GENERATION DNA SEQUENCER

使用微加工设备进行DNA测序有很多优点；其中许多都来自芯片注入器。举几个例子，(1)注入一个非常窄的频带，(2)获得均匀的信号强度分布图，(3)将样本量减少到亚mul，(4)进行额外的样品清理。然而，微加工设备有两个主要的限制。(a)直道长度有限，(b)储层布置空间有限。提出了一种基于芯片注入器与传统毛细管耦合的新一代DNA测序仪。从芯片测序的角度来看，该方案消除了这两个限制。由于任何长度的毛细管都可以耦合到切屑注入器上，因此直道长度变得无限，并且由于毛细管的灵活性，可以使用许多单独的切屑注入器来容纳储层，因此储层的空间也变得无限。从使用CGE测序的角度来看，该方案将芯片进样器的所有优点都引入了CGE。测序仪将包括一个4-PMT旋转共聚焦扫描仪与8x48毛细管阵列。线性聚丙烯酰胺将被优化用于测序仪上的测序分离。将纳入小体积样品制备以减少样品体积，并将开发自动化仪器以提高样品吞吐量。通过与基因组测序中心（如Genome Therapeutics Corporation）的密切合作，将开发出满足基因组测序世界需求的测序仪。新一代测序仪将具有24小时最少注意操作的规格，平均读取长度为800-1000个碱基，运行时间小于1小时，样品体积为0.5至1 μ l。

DNA sequencing using microfabricated devices has a lot of advantages; many of them are originated from a chip injector. To list a few, (1) a very narrow band is injected, (2) a uniform signal intensity profile is obtained, (3) sample volume is reduced to sub-muL and (4) extra sample cleanup is performed. However, there are two major limitations associated with microfabricated devices. (a) limited straight channel length and (b) constrained space for reservoir arrangement. A new generation DNA sequencer based a chip injector coupled with conventional capillaries is proposed. Viewed from the perspective of sequencing on a chip, the proposed scheme eliminates the two limitations. Straight channel length becomes unlimited since a capillary of any length can be coupled to a chip injector and the space for reservoirs becomes unlimited as well since many separate chip injectors can be used to host reservoirs due to the flexibility of the capillaries. Seen from the perspective of sequencing using CGE, the proposed scheme brings all advantages of a chip injector to CGE. The sequencer will consist of a 4-PMT rotary confocal scanner with 8x48-capillary arrays. Linear polyacrylamide will be optimized for sequencing separation on the proposed sequencer. Small volume sample preparation will be incorporated to reduce the sample volume and automated instrumentation will be developed to enhance the sample throughput. Through close collaboration with Genome Sequencing Centers (e.g. Genome Therapeutics Corporation), the sequencer will be developed to meet the needs of genome sequencing world. The new generation sequencer will have specifications of 24-hour operation with minimum attention, average read-length of 800-1000 bases, a run-to-run time of less than 1 hour, and a sample volume of 0.5 to 1 muL.