DNA Sequencing

DNA测序

• 批准号: 8732229
• 负责人: EDWARD E. PARTRIDGE
• 金额: $ 13万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-03-28 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8732229
• 关键词: Accounting; Acquired Immunodeficiency Syndrome; Automation; Back; Base Pairing; Basic Science; Budgets; Cancer Center; Cancer Vaccines; Capillary Electrophoresis; Cellular biology; Charge; Comprehensive Cancer Center; Contracts; DNA; DNA Sequence; Development; Dideoxy Chain Termination DNA Sequencing; Ensure; Equipment; Genetic; Genomics; Goals; Grant; Health Services Accessibility; Immunology; Instruction; Laboratories; Malignant Neoplasms; Management Information Systems; Methodology; Microsatellite Repeats; Preparation; Process; Reaction; Research; Research Personnel; Sequence Analysis; Services; System; Technology; Time; Training; Training Support; Translational Research; University of Alabama at Birmingham Cancer Center; cancer therapy; cost; cost effective; design; member; programs; success; tool

The goal of the DNA Sequencing Shared Facility (DSSF) is to provide UAB Comprehensive Cancer Center (CCC) members with state-of-the-art automated Sanger sequencing. To this end, the specific aims are: 1. To continue to provide state-of-the-art automated DNA sequencing capabilities by maintaining dedicated Applied Biosystems Sequencers (ABI 3730XL) with capillary electrophoresis systems. 2. To continue to provide technical support and training to Cancer Center members, especially junior investigators, in DNA template preparation, primer design, cycle sequencing methodologies, and standard DNA sequence editing and analysis. To expand facility functions by providing microsatellite analyses and real-time PCR technologies as new services. To effectively interface with the Microarray Shared Facility (MSF) and the Comprehensive Genomics Shared Facility (CGSF) to assure services are complementary and not overlapping. Although originally established within the Center for AIDS Research (in 1987), CCC members now account for 70% of this facility's usage. This is because numerous laboratories within the Cancer Center are absolutely dependent on high throughput DNA sequencing for their research programs. The DSSF has ensured that CCC members have access to this service in a timely and cost-effective manner. In the last budget period, the DSSF has developed new methodologies for the automated sequencing process, trained CCC investigators in template preparation and the use of sequence analysis tools, upgraded and modernized pertinent equipment, employed automation technologies to expand capacity, and installed a Laboratory Information Management System to optimize the user interface. Since 2004, the DSSF has supported 104 CCC investigators and provided essential services for more than 175 cancer and cancer related grants and contracts. More than 458 million base pairs of primary sequence were determined, generating more than $2,710,000 in user charge-backs. Newly implemented automation increased the facility's efficiency and allowed the DSSF to reduce user charge-backs from $8 to $6 per sequencing reaction ($4 per reaction for 96 well plates).

DNA测序共享设施（DSSF）的目标是提供UAB综合癌症中心 (CCC)最先进的自动化桑格测序。为此，具体目标是： 1.继续提供最先进的自动化DNA测序能力， 专用Applied Biosystems测序仪（ABI 3730XL），配有毛细管电泳系统。 2.继续为癌症中心成员提供技术支持和培训，特别是初级 研究人员，在DNA模板制备，引物设计，循环测序方法， 标准DNA序列编辑和分析。 通过提供微卫星分析和实时PCR技术， 新服务。 为了有效地与微阵列共享设施（MSF）和综合基因组学 共享设施（CGSF），以确保服务是互补的，而不是重叠。 虽然最初成立于艾滋病研究中心（1987年），CCC成员现在占 这座设施70%的使用量这是因为癌症中心内的许多实验室 他们的研究项目完全依赖于高通量DNA测序。DSSF已 确保CCC成员能够以及时和具有成本效益的方式获得这项服务。在过去 在预算期间，DSSF已经开发了自动排序过程的新方法， CCC研究人员在模板制备和序列分析工具的使用方面， 对相关设备进行了现代化改造，采用自动化技术扩大了产能， 优化实验室信息管理系统的用户界面。自2004年以来，DSSF 为104名CCC调查人员提供支持，并为超过175名癌症和癌症患者提供基本服务 相关赠款和合同。超过4.58亿个碱基对的一级序列被确定， 产生了超过271万美元的用户退款。新实施的自动化增加了 提高了设施的效率，并使DSSF能够将每次测序的用户收费从8美元减少到6美元 反应（96孔板每个反应4美元）。