MICROARRAY FABRICATION, HYBRIDIZATION, ANALYSIS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 微阵列核心设备(MCF)提供了一种同时测量数千个基因表达的专门方法。这在表征对疾病、有毒侮辱和其他外部刺激的分子反应方面非常有用。MCF拥有大量的遗传信息库，可用于识别参与此类反应的基因。这使我们能够通过信使RNA的相对水平来测量一张显微镜载玻片上数万个基因的表达。我们为用户提供实验计划和定制的微阵列打印(如果需要)，然后继续进行标准微阵列实验的步骤。RNA被分离和纯化，然后转化为更稳定的用荧光染料标记的cDNA.用专门的激光扫描显微镜在显微镜载玻片上和对照样品上测量这一cdna。然后对数据进行分析，以阐明在基因水平上对给定刺激的反应。然后，这些信息被用来确定可能正在发生的某些细胞过程或整个组织反应。这最终可能导致识别对疾病或毒素有反应的基因的“特征”，提供更早检测或治疗机制的方法。这是一个强大的工具，因为它不需要可能限制调查人员研究范围的特定假设。它被认为是“假设生成”而不是“假设驱动”实验。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Microarray Core Facility (MCF) provides a specialized method of measuring the expression of thousands of genes simultaneously. This can be extremely useful in characterizing the molecular reactions to diseases, toxic insults and other external stimuli. The MCF has large libraries of genetic information available to identify the genes involved in such reactions. This gives us the capability of measuring the expression of tens of thousands of genes on a single microscope slide by the relative levels of messenger RNA. We provide users with experiment planning and custom microarray printing (if necessary) then proceed through the steps of a standard microarray experiment. RNA is isolated and purified, then converted to more stable cDNA that is labeled with fluorescent dye. This cDNA is measured on a microscope slide with a control sample by a specialized laser scanning microscope. The data is then analyzed to elucidate the response to the given stimulus at the genetic level. This information is then used to determine certain cellular processes or whole tissue responses that may be occurring. This can eventually lead to the identification of a "signature" of genes that respond to a disease or toxin, providing methods of earlier detection or mechanisms of treatment. This is a powerful tool in that it does not require a specific hypothesis that may restrict the scope of an investigator's research. It is considered "hypothesis generating" rather than "hypothesis driven" experimentation.