MICROARRAY FABRICATION, HYBRIDIZATION, ANALYSIS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 微阵列核心设施（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、毒性损伤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的相对水平来测量单个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自定义微阵列打印（如有必要），然后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cDNA，用荧光染料标记。 该cDNA通过专用激光扫描显微镜在具有对照样品的显微镜载玻片上测量。 然后对数据进行分析，以阐明在遗传水平上对给定刺激的反应。 然后，该信息用于确定可能发生的某些细胞过程或整个组织反应。 这最终可以导致识别对疾病或毒素作出反应的基因的“特征”，从而提供早期检测或治疗机制的方法。 这是一个强有力的工具，因为它不需要一个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”的实验。

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.