Core--Analysis

描述（由申请人提供）： 分析中心将为基因治疗相关的分子分析方法以及细胞和组织对基因转移的反应提供专业知识、人员和设备。分析中心位于新装修的Belfer基因治疗中心设施内，由实验室空间和设备室组成，其中包含序列检测系统7700（TaqMan）实时定量PCR系统和BioMek FX样品制备机器人。该核心将向该计划项目的研究人员提供与基因治疗相关的几种先进分子技术。其中包括：自动DNA提取和纯化;用于DNA定量的TaqMan定量PCR和用于评估基因表达水平变化的定量RT-PCR。虽然不是计划项目中拟议研究的特定部分，但也可获得高通量基因分型、基因表达微阵列（Affyscore GeneChip）以及基因组和微阵列表达数据分析。所有四个项目都将平等地使用分析核心，主要用于定量PCR领域，用于定量细胞和组织中的载体DNA，以及定量RTPCR领域，用于基因表达水平的绝对和相对定量。分析中心主任Neil Hackett花了大量时间优化定量PCR（TaqMan）方法来评估基因转移，并证明了该方法的潜在能力和高通量， 正确执行。由于分子技术的复杂性和昂贵性以及所需的先进设备，任何一个团队都难以获得通过分析核心获得的高通量所带来的所有复杂方法和效率。

DESCRIPTION (provided by applicant): The Analysis Core will provide expertise, personnel and equipment for molecular analysis methods relevant to gene therapy and the response of cells and tissues to gene transfer. The Analysis Core is located in the newly renovated Belfer Gene Therapy Core Facility and consists of laboratory space and an equipment room containing the Sequence Detection System 7700 (TaqMan) realtime quantitative PCR system and BioMek FX sample preparation robot. The Core will make available to the investigators of the Program Project several advanced molecular techniques relevant to gene therapy. These include: automated DNA extraction and purification; TaqMan quantitative PCR for DNA quantitation and quantitative RT-PCR for assessment of changes in gene expression levels. Also available, although not specifically part of the proposed studies in the Program Project, are high throughput genotyping, gene expression microarrays (Affymetrix GeneChip), and analysis of genomic and microarray expression data. All four projects will make equal use of the Analysis Core, primarily in the area of quantitative PCR for quantitation of vector DNA in cells and tissues and in quantitative RTPCR for the absolute and relative quantitation of gene expression levels. The Director of the Analysis Core, Neil Hackett, has spent considerable time optimizing quantitative PCR (TaqMan) methods to assess gene transfer and demonstrated the potential power and high throughput of the method when properly executed. Due to the complexity and expense of molecular techniques and the advanced equipment required, it is a challenge for any one group to acquire all of the sophisticated methodologies and efficiency derived from high throughput that are available through the Analysis Core.