Roche Genome Sequencer 20 System

描述（由申请人提供）：在这个共享仪器补助金，我们正在寻求资金购买罗氏基因组测序仪20（GS 20）。该仪器基于454 Life Sciences开发的革命性测序技术，现在通过Roche Diagnostics分销。在9月的一篇自然文章中描述。2005 [1]，GS 20在单次4.5小时运行中提供了大约二十万个100个碱基对的序列读数，试剂和供应品的成本略高于6，000美元。也就是说，可以获得20 Mbp的序列，样品制备2-3天，成本比基于标准电泳的技术低100倍（基于每bp）。该技术为高通量发现提供了全新的机会，其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，在华盛顿大学和其他地方都存在大量具有不同致病性的临床分离株。虽然存在大量的分子生物学技术，人们可以使用这些技术来寻找与表型相关的基因组区域，但所有目前的方法都是耗时且繁琐的。细菌基因组的廉价测序将能够更有效地鉴定和表征与临床结果相关的病原体序列特征。2)逆转录病毒群体内序列分布的表征。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。这种测序技术将被应用于更快地了解病毒进化，这将有助于疫苗和治疗方法的开发。3)哺乳动物基因组中选定基因组区域或cDNA的快速重新测序。通过对目前公开的方案进行适度修改，该技术将适用于cDNA库的高通量全长测序和群体中选定基因的深度再测序以鉴定罕见多态性。

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.