GENE EXP. AND GENOTYPING

基因表达和基因分型设施的任务是促进HIGH的实施 由案例社区成员进行的研究的吞吐量基因技术。全球创业投资基金 为癌症中心成员以及其他研究人员提供技术和咨询资源 在凯斯西储大学、克利夫兰大学医院、克利夫兰诊所拥有实验室 基金会、退伍军人医院和MetroHealth医院在高通量表达分析和 基因分型。这是它在克利夫兰唯一有能力的设施。该设施利用Affymetrix和应用 生物系统(ABI)技术，提供全球和定量(根据用户识别的基因)基因表达 分析服务。多种Affymetrix和ABI技术可用于解决 基因分型需求，包括全基因组扫描，定制设计的精细图谱研究，涉及3,000-10,000 SNPs和对数千个样本中少量(&lt；100)特定SNPs的询问。核心运行在 全方位服务模式，由工作人员提供初步咨询、执行化学分析、以及 提供数据修剪和数据解释服务。为了实施这些技术，GEGF有14个 计算机、1个专门用于数据生成的Affymetrix工作站、3个流体站、2个杂交炉和 带自动加载机的GC3000扫描仪。ABI设备包括一个7900序列检测系统，带有条形码 阅读器、机器人技术，能够每天处理5,000个样品井，以及用于处理的模块 低密度阵列，96孔板和384孔板。GEGF还拥有5台GeneAMP ABI 9700聚合酶链式反应机器，其中96台- &384井阻断能力和ABI 6100核酸预备站。对于基于聚合酶链式反应的分析，GEGF 建立了一个无扩增子的房间，里面有2个聚合酶链式反应引擎盖、专用移液器、小型微熔断器和微型热循环器。 其他设备包括：2个矩阵机器人液体处理系统，PlateMate 2x2(X-Y 平台移动)和PlateMate Plus(允许从96孔格式到384孔格式的快速重新配置)，专用 离心机、安捷伦生物分析仪和NanoDrop分光光度计，用于定量1-2微升的核酸 样本。样品准备和分析的周转时间对于一般规模的项目和 在初步协商期间，为大型项目制定了令人满意的时间表。癌症中心成员 享受GEGF提供的所有服务的35%折扣。核心从2000年成立以来一直在稳步增长， 当我们有22名用户和一名全职工作人员时，它的现有工作人员为51/2人，注册用户为142人。 每年大约有60名研究人员使用GEGF，其中30%-50%是第一次使用，记录 政府间常设论坛继续享有健康的增长水平和与回头客的持续工作的强劲水平。 最后，GEGF为癌症中心的8个项目的成员提供了服务，并发挥了 在获得授予癌症中心成员的33多笔赠款方面发挥了重要作用。

The mission of the Gene Expression and Genotyping Facility (GEGF) is to facilitate implementation of high throughput genetic technologies for the research conducted by members of the Case community. The GEGF serves as a technical and consulting resource for Cancer Center members, as well as other investigators who have laboratories at Case Western Reserve University, University Hospitals of Cleveland, the Cleveland Clinic Foundation, the VA Hospital, and MetroHealth Hospital in the areas of high throughput expression analysis and genotyping. It is the only facility of its capability in Cleveland. This facility utilizes Affymetrix and Applied Biosystems (ABI) technologies to provide global and quantitative (on user-identified genes) gene expression analysis services. A diverse array of Affymetrix and ABI technologies are available to address a spectrum of genotyping needs, including whole genome scans, custom designed fine-mapping studies involving 3,000-10,000 SNPs and interrogation of small numbers (<100) of specific SNPs in thousands of samples. The core operates in a full-service mode with staff providing initial consultation, executing the chemistry, carrying out the analysis, and offering data trimming and data interpretation services. To carry out these technologies, the GEGF has 14 computers, an Affymetrix Workstation dedicated to data generation, 3 fluidics stations, 2 hybridization ovens and a GC3000 scanner with autoloader. ABI equipment includes a 7,900 Sequence Detection System, with a barcode reader, robotics allowing the capability to process 5,000 sample wells per day, and the modules for processing Low Density Arrays, 96- and 384-well plates. The GEGF also has 5 GeneAMP ABI 9700 PCR machines, with 96- & 384-well block capability and an ABI 6100 Nucleic Acid Prep Station. For PCR-based assays, the GEGF established an amplicon-free room, containing 2 PCR hoods, dedicated pipettors, small microfuges and minithermocyclers. Additional equipment includes: 2 Matrix robotic liquid handling systems, the PlateMate 2x2 (X-Y stage movement) and the PlateMate Plus (allows rapid reconfiguration from 96- to 384-well format), dedicated centrifuges, an Agilent Bioanalyzer and a NanoDrop Spectrophotometer, which quantifies nucleic acids in 1-2 ul samples. Turn-around time for sample preparation and analysis is <10 days for average size projects and satisfactory timetables are established for large projects during initial consultations. Cancer Center members receive a 35% discount on all GEGF provided services. The core has steadily grown from its inception in 2000, when we had 22 users and a single full-time staff member to its current staff of 51/2 and 142 registered users. Approximately 60 investigators use the GEGF every year and 30-50% of these are first time users, documenting that the GEGF continues to enjoy a healthy level of growth and a robust level of sustained work with repeat users. Finally, the GEGF has provided services for members from 8 of the Cancer Center's programs, and has played a role in securing more than 33 grants awarded to Cancer Center members.