Roche Genome Sequencer 20 System

描述(由申请人提供)：在这项共享仪器赠款中，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，目前通过罗氏诊断公司分销。在9月1日发表在《自然》杂志的一篇文章中描述了这一点。2005[1]，GS20在一次4.5小时的运行中提供了大约200,100个碱基对序列读数，而试剂和用品的成本略高于6,000美元。也就是说，人们可以通过2-3天的样品制备获得20Mbp的序列，而成本(以每个bp为基础)比基于标准电泳法的技术低100倍。这项技术为高通量发现提供了全新的机会，最初将在华盛顿大学的以下领域应用：1)对临床分离株和以前测序的细菌基因组的表型感兴趣的菌株进行测序。对于许多致病细菌，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。虽然有大量的分子生物学技术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和繁琐的。廉价的细菌基因组测序将使更有效地识别和表征与临床结果相关的病原体序列特征。2)逆转录病毒种群内序列分布的特征。逆转录病毒基因组在宿主体内迅速变异，对宿主防御和治疗做出反应。这项测序技术将被应用于更快速地了解病毒的进化，这将有助于疫苗和治疗的开发。3)对哺乳动物基因组中选定的基因组区域或cDNAs进行快速重新测序。通过对目前公布的方案进行适当的修改，这项技术将适用于对cDNA池进行高通量、全长测序，并对种群中选定的基因进行深度重新测序，以识别罕见的多态。

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.