AB3500xL DNA Analyzer for OHSU Core Facility

描述（由申请人提供）：应用生物系统/生命技术3500 xL（24毛细管）DNA分析仪的资金要求。它将成为DNA服务核心设施（www.ohsu.edu/xd/research/research-cores/dna- analysis/）提供的DNA测序和片段分析的主要仪器。这一完善的核心设施为23个OHSU部门、中心和研究所以及其他12个非OHSU波特兰地区部门的约100名主要研究人员提供毛细管电泳测序服务。长读段桑格测序是所有进行现代假设驱动的分子生物学实验室所需的基本技术。预计大规模并行的全基因组研究将在近期增加后续桑格测序的需求。今年我们将运行约30，000个样本。有了所要求的仪器，我们将能够使生产率翻一番。在我们支持的200多个NIH资助的基础和转化项目中，有几个是奈特癌症研究所Brian Druker实验室的“加速白血病的靶点识别和验证”;俄勒冈州干细胞中心Markus Grompe实验室的“远端酪氨酸途径疾病的治疗”;以及耳鼻喉科医学院Teresa Nicolson实验室的“毛细胞功能的遗传和分子解剖”。我们的核心测序项目包括分子微生物学和免疫学、传染病、眼科、细胞生物学和发育以及医学院、职业与环境毒理学研究中心（CROET）和波特兰州立大学生物学系的肾脏病学等部门，这些部门的测序服务使我们的主要用户群体更加完善，占我们当前使用量的55%以上。该补助金的主要参与者设计并实施了核心用于接收工作请求、跟踪工作以及履行计费和报告职能的数据库和网站。除了Keller博士之外，技术人员还包括一名具有多年自动DNA测序经验的全职技术人员和一名半职技术人员。当前的16毛细管3130 xL测序仪将用于更短、更快的PCR测序运行;并用于维护和维修停机期间的备份。要求的24毛细管仪器将用于需要长读数（800-1000个碱基）和片段分析的样本。所要求的仪器将显着提高我们的数据吞吐量和基因组分析的生产力。

DESCRIPTION (provided by applicant): Funds for an Applied Biosystems / Life Technology 3500xL (24 capillary) DNA Analyzer are requested. It will become the principle instrument for the DNA sequencing and fragment analysis provided by the DNA Services Core Facility (://www.ohsu.edu/xd/research/research-cores/dna- analysis/). This well-established core facility provides capillary electrophoresis sequencing services to ~100 principle investigators in 23 OHSU departments, center and institutes, and 12 other non-OHSU Portland area departments. Long-read Sanger sequencing is a basic technique required by all labs performing modern, hypothesis-driven molecular biology. Massively parallel whole genome research is expected to increase the need for follow-up Sanger sequencing in the near term. We will run ~30,000 samples this year. With the requested instrument, we will be able to double that productivity. A few, of the more than 200 basic and translational NIH funded projects we support, are "Accelerating target identification and validation in leukemia" in the Brian Druker lab in the Knight Cancer Institute; "Treatment of Distal Tyrosine pathway Disorders" in the Markus Grompe lab in the Oregon Stem Cell Center; and "Genetic and molecular dissection of hair-cell function" in the Teresa Nicolson lab in the School of Medicine, Otolaryngology Research Department. Sequencing by our core for projects in Molecular Microbiology & Immunology, Infectious Disease, Ophthalmology, Cell Biology & Development and Nephrology departments in the School of Medicine, the Center for Research on Occupational & Environmental Toxicology (CROET) and the Biology Department at Portland State University round out our major user group; accounting for >55% of our current usage. The PI for this grant designed and implemented the databases and website used by the core for receiving work-requests, tracking work and performing billing and reporting functions. The technical staff, besides Dr. Keller, consists of one full-time technician with several years of automated DNA sequencing experience and one-half time technician. The current, 16 capillary 3130xL sequencer will be used for shorter, faster PCR sequencing runs; and for backup during maintenance and repair downtime. The requested 24 capillary instrument will be used for samples requiring the long reads (800-1000 bases) and fragment analysis. The requested instrumentation will significantly increase our data throughput and genomic analysis productivity.