GENOTYPING

基因分型

• 批准号: 8039947
• 负责人: QING WANG
• 金额: $ 20.64万
• 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/8039947
• 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; 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的目的是作为一个集中的设施进行大规模和高通量基因分型 基因多态性以及常规DNA测序。这种集中式设施的优点是 包括：1）节约资源;核心实验室比单个实验室更节省时间和成本; 2）避免 不必要的重复工作：3）质量控制标准化; 4）促进数据共享。在 此外，该核心还将作为一种教育资源，对计划项目的成员进行培训 实验室在适当的基因分型方法，并提供建议，为项目负责人在设计和 进行相关实验。培训将是宝贵的博士后研究员和研究生 学生和有用的，如果对核心的需求超过了它的能力，在特定的时间.基因分型 核心将提供一台ABI Prism 7900 HT遗传分析仪（配备SDS 384孔板）和一台半自动 机器人系统，Hydra II用于高通量SNP基因分型。我们将使用5'核酸酶等位基因 用于SNP的大规模、高通量基因分型的鉴别测定。所有SNP检测均将检测 其准确性和可重复性。我们已经制定了严格的方案，用于评估每个SNP检测 其中通过Wave系统或直接测序鉴定感兴趣的SNP。SNP选自 已发表的数据库将首先通过直接测序进行确认，然后进行稳健性测试， 测定的重现性。如果SNP通过这些测试，它将在其余的研究样本中进行基因组化。 为了进一步的质量控制，我们包括两个阴性对照（无DNA），两个等位基因1标准品的威尔斯孔，和两个等位基因1标准品的孔。 等位基因2标准品的威尔斯孔。基因分型核心是该计划项目的一个组成部分。项目 1和2将广泛使用SNP检测服务。其他项目将利用DNA 用于分析质粒探针和cDNA克隆的测序设施（项目3）。