DEVELOPMENT OF A NEW GENERATION DNA SEQUENCER

使用微型制造设备进行DNA测序有很多优点；其中许多来自芯片注入器。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，(3)将样本体积减少到亚微米，(4)执行额外的样本清理。然而，与微制造器件相关的主要限制有两个。(A)有限的直航道长度和(B)有限的水库布置空间。提出了一种基于芯片注入器和常规毛细管相结合的新一代DNA测序仪。从片上排序的角度来看，该方案消除了这两个限制。直槽长度变得无限，因为任何长度的毛细管可以耦合到芯片注入器，并且储集层的空间也变得无限，因为由于毛细管的灵活性，许多单独的芯片注入器可以用来容纳储集层。从CGE测序的角度来看，该方案将芯片注入器的所有优点都带到了CGE中。该测序仪将由一个带有8x48毛细管阵列的4-PMT旋转共聚焦扫描仪组成。线性聚丙烯酰胺将在建议的测序仪上优化用于测序分离。将采用小体积样品制备，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(如基因组治疗公司)的密切合作，该测序仪将被开发出来，以满足基因组测序世界的需求。新一代测序仪的规格为24小时最小关注操作，平均读取长度为800-1000个碱基，运行到运行时间不到1小时，样本量为0.5-1-1。

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.