Roche Genome Sequencer 20 System

描述（由申请人提供）：在这项共享仪器拨款中，我们正在寻求资金购买罗氏基因组测序仪20 （GS20）。该仪器基于454生命科学公司开发的革命性测序技术，现在通过罗氏诊断公司销售。在2005年9月《自然》杂志的一篇文章中，GS20在一次4.5小时的运行中提供了大约20万个100个碱基对序列的读取，而试剂和耗材的成本略高于6000美元。也就是说，可以获得20Mbp的序列，2-3天的样品制备成本（以每bp为基础）比基于标准电泳的技术低100倍。这项技术为高通量发现提供了全新的机会，将首先在华盛顿大学应用于以下领域：1)对先前测序的细菌基因组的临床分离株和表型感兴趣的菌株进行测序。对于许多致病菌，华盛顿大学和其他地方都有大量具有不同致病性的临床分离菌株。虽然存在大量的分子生物学技术，人们可以用来寻找与表型相关的基因组区域，但目前所有的方法都是耗时且乏味的。廉价的细菌基因组测序将使与临床结果相关的病原体序列特征的识别和表征更加有效。2)逆转录病毒群体序列分布的表征。逆转录病毒基因组在宿主体内迅速突变，以响应宿主的防御和治疗。这种测序技术将用于更快地了解病毒的进化，这将有助于开发疫苗和治疗方法。3)哺乳动物基因组中选定的基因组区域或cDNA的快速重测序。通过对目前已发表的协议进行适度修改，该技术将适用于高通量、cDNA库的全长测序和种群中选定基因的深度重测序，以识别罕见的多态性。

DESCRIPTION (provided by applicant): In this shared instrumentation grant, we are seeking funds to purchase a Roche Genome Sequencer 20 (GS20). This instrument is based on the revolutionary sequencing technology developed at 454 Life Sciences and now distributed through Roche Diagnostics. Described in a Nature article in Sept. 2005 [1], the GS20 provides approximately two hundred thousand, 100 base pair sequence reads in a single 4.5hr run at a cost of a little over $6,000 in reagents and supplies. That is, one can obtain 20Mbp of sequence, with 2-3 days of sample preparation at a cost that is 100 fold (on a per bp basis) lower than standard electrophoresis based technologies. This technology provides radical new opportunities for high throughput discovery which will be initially applied at the University of Washington in the following areas: 1) Sequencing of clinical isolates and phenotypically interesting strains of previously sequenced bacterial genomes. For many pathogenic bacteria, large collections of clinical isolates with varying pathogenicity exist both at the University of Washington and elsewhere. While a large number of molecular biological techniques exist that one can use to hunt for genomic regions that are correlated with phenotype, all present day methods are time consuming and tedious. Inexpensive sequencing of bacterial genomes will enable a far more efficient identification and characterization of pathogen sequence traits that correlated with clinical outcomes. 2) Characterization of sequence distributions within retroviral populations. Retroviral genomes rapidly mutate within the host in response to both the host defenses and therapy. This sequencing technology will be applied to a gaining a more rapid understanding of viral evolution that will aid in the development of vaccines and treatments. 3) Rapid re-sequencing of selected genomic regions or cDNA's in mammalian genomes. With modest modifications to currently published protocols, this technology will be adapted to high throughput, full length sequencing of pools of cDNA's and to deep re- sequencing of selected genes in a population to identify rare polymorphisms.