CORE--DNA SEQUENCING

核心——DNA测序

• 批准号: 6479991
• 负责人: THOMAS Martin HERING
• 金额: $ 3.83万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6479991
• 关键词: DNA; biomedical facility; family genetics; human genetic material tag; human tissue; nucleic acid sequence; osteoarthritis; tissue resource /registry

The DNA sequencing core facility has been an important component of the NEOMAMDC since 1993, and has significantly enhanced the research infrastructure of the Center, and accelerated progress for Arthritis researchers. Although the scope of molecular biology procedures used by laboratories associated with the Arthritis Center is quite broad and are project-specific most laboratories of the NEOMAMDC have benefitted and will continue to benefit from a core facility dedicated to DNA sequence determination. The rationale for this is that 1) DNA sequencing is required at some point in any molecular biology oriented experimental approach, and 2) it is not always cost effective for each laboratory to set up individual protocols, purchase equipment an train the necessary personnel. 3) DNA sequence analysis is labor and time intensive, but does not require specific knowledge of the large project. 4) The on site sequencing facility enhances the overall productivity of Arthritis Center research laboratories by performing this task more efficiently and with greater precision than would be possible on an individual basis 5) The core facility frees time for investigators to engage in more creative pursuits. The primary objective of the core is to assist those investigators already engaged in molecular biology protocols. In order to be practical, it does not serve to perform any molecular biology technique other than DNA sequence analysis. Investigators are responsible for preparation of samples for sequencing in advance of submission. Protocols for samples preparation are provided by the sequencing core in order to assure that the core facility will be efficiently utilize and to assure success. In general, investigators are responsible for ligation of DNA fragments into the polylinker (cloning site) of sequencing vectors commonly in use. The core facility is then responsible for performing the sequencing reactions, running the automated sequencing instrument and recording the sequence as a file that can be used for further analysis by the investigator. The second objective of the core is to serve as repository for genomic DNA from members of families afflicted with OA showing familial inheritance, and from patients with OA disease subsets. DNA from these individuals is made available to other investigators on a collaborative basis. Additionally, the core functions to screen this DNA bank for mutations in the type II collagen gene.

DNA测序核心设施已成为重要组成部分 NEOMAMDC 自 1993 年以来，显着加强了研究 中心的基础设施，并加速关节炎的进展 研究人员。尽管分子生物学程序的范围 与关节炎中心相关的实验室非常广泛， NEOMAMDC 的大多数实验室都已受益于特定项目 将继续受益于 DNA 序列专用核心设施 决心。这样做的理由是 1) DNA 测序是 在任何以分子生物学为导向的实验中的某个时刻都需要 方法，2) 每个实验室并不总是具有成本效益 制定单独的协议、购买设备并培训必要的人员 人员。 3) DNA 序列分析是劳动和时间密集型的，但 不需要大型项目的具体知识。 4）现场 测序设施提高了关节炎中心的整体生产力 研究实验室通过更有效地执行这项任务 比单独的精度更高 5) 核心设施为调查人员腾出时间从事更具创造性的工作 的追求。 核心的主要目标是协助那些已经 从事分子生物学协议。为了实用，这样做 不用于执行除 DNA 之外的任何分子生物学技术 序列分析。调查人员负责准备 在提交之前进行测序的样品。样品协议 准备工作由测序核心提供，以确保 核心设施将得到有效利用并确保成功。在 一般来说，研究人员负责将 DNA 片段连接到 常用的测序载体的多接头（克隆位点）。这 然后核心设施负责执行测序反应， 运行自动测序仪器并将序列记录为 调查人员可用于进一步分析的文件。 核心的第二个目标是作为基因组 DNA 的储存库 来自患有 OA 的家庭成员，显示出家族遗传， 以及来自 OA 疾病亚型的患者。这些人的 DNA 是 在协作的基础上提供给其他研究人员。 此外，该 DNA 库的核心功能是筛选突变 II型胶原蛋白基因。