GENOTYPING

基因分型

• 批准号: 7799808
• 负责人: QING WANG
• 金额: $ 20.03万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7799808
• 关键词: Alleles; Biological Assay; Cloning; DNA; DNA Sequence; DNA Sequencing Facility; Detection; Discrimination; Evaluation; Genetic; Genetic Polymorphism; Genotype; Hydra Polyps; Individual; Laboratories; Methodology; Plasmids; Postdoctoral Fellow; Protocols documentation; Published Database; Quality Control; Reproducibility; Resources; Rest; Robotics; SNP genotyping; Sampling Studies; Services; Standardization; System; Testing; Thrombosis; Time; Training; cDNA Probes; cost efficient; data sharing; design; graduate student; interest; member; nuclease; programs; research study; Alleles; Biological Assay; Cloning; DNA; DNA Sequence; DNA Sequencing Facility; Detection; Discrimination; Evaluation; Genetic; Genetic Polymorphism; Genotype; Hydra Polyps; Individual; Laboratories; Methodology; Plasmids; Postdoctoral Fellow; Protocols documentation; Published Database; Quality Control; Reproducibility; Resources; Rest; Robotics; SNP genotyping; Sampling Studies; Services; Standardization; System; Testing; Thrombosis; Time; Training; cDNA Probes; cost efficient; data sharing; design; graduate student; interest; member; nuclease; programs; research study

The purpose of Core D is act as a centralized facility to perform large scale and high throughput genotyping of genetic polymorphisms as well as routine DNA sequencing. The advantages for such a centralized facility include: 1) Conservation of resources; the core is more time and cost efficient than individual labs; 2) Avoid unnecessary duplication of effort: 3) Standardization of quality control; and 4) Facilitation of data sharing. In addition, the Core will also function as an educational resource to train members of the Program Project laboratories in proper genotyping methodologies and to provide advice for Project Leaders in the design and implementation of relevant experiments. The training will be valuable for Postdoctoral Fellows and Graduate Students and useful if the demands on the Core exceed its capacity at a particular time. The Genotyping Core will provide one ABI Prism 7900HT Genetic Analyzer with SDS 384-well Block, and a semi-automatic robotic system, Hydra II for high throughput SNP genotyping. We will use the 5' nuclease allelic discrimination assay for large-scale, high throughput genotyping of SNPs. All SNP assays will be tested for their accuracy and reproducibility. We have developed a strict protocol for evaluation of each SNP assay where the SNP of interest is identified by the Wave system or direct sequencing. SNPs chosen from published databases will be first confirmed by direct sequencing, followed by testing for the robustness and reproducibility of the assay. If the SNP passes these tests, it will be genotyoed in the rest of study samples. For further quality control, we include two negative controls (no DNA), two wells of allele 1 standard, and two wells of allele 2 standard. The Genotyping Core represents an integral part of the Program Project. Projects 1 and 2 will use the SNP detection service extensively. The other projects will make use of the DNA sequencing facility for analysis of plasmid probes and cDNA cloning (Project 3).

核心D的目的是作为进行大规模和高通量基因分型的集中式设施 遗传多态性以及常规的DNA测序。这种集中设施的优势 包括：1）资源保护；核心比单个实验室更有时间和成本效益。 2）避免 不必要的努力重复：3）质量控制的标准化； 4）促进数据共享。在 此外，核心还将作为培训计划项目成员的教育资源 实验室进行适当的基因分型方法，并为设计领导者提供建议 实施相关实验。该培训对于博士后研究员和毕业生将很有价值 学生，如果对核心的需求超过其在特定时间的能力，则有用。基因分型 核心将提供一个ABI Prism 7900HT遗传分析仪，SDS 384孔盖和一个半自动 机器人系统，高通量SNP基因分型的Hydra II。我们将使用5'核酸酶等位基因 SNP的大规模，高通量基因分型的歧视分析。所有SNP分析将进行测试 它们的准确性和可重复性。我们已经开发了一个严格的协议来评估每个SNP分析 其中关注的SNP通过波系统或直接测序识别。从中选择的SNP 已发布的数据库将首先通过直接测序确认，然后测试鲁棒性和 测定的可重复性。如果SNP通过这些测试，则将在其余的研究样本中进行基因诺。 为了进一步的质量控制，我们包括两个负面对照（无DNA），等位基因1标准的两个井和两个 等位基因2标准的井。基因分型核心代表了程序项目的组成部分。项目 1和2将广泛使用SNP检测服务。其他项目将利用DNA 测序设施用于分析质粒探针和cDNA克隆（项目3）。