CORE--BRAIN MICRODISSECTION, RNA AND PROTEIN ANALYSIS

核心——脑显微切割、RNA和蛋白质分析

• 批准号: 6234440
• 负责人: BRADLEY T. HYMAN
• 金额: $ 14.55万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-15 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6234440
• 关键词: RNA; RNase protection assay; analytical method; animal genetic material tag; autoradiography; biomedical facility; densitometry; dissection; genetic registry /resource /referral center; human genetic material tag; human tissue; nucleic acid chemical synthesis; nucleic acid probes; polymerase chain reaction; postmortem; synthetic nucleotide; tissue /cell preparation; tissue resource /registry; western blottings

This core will provide a central resource for reagent synthesis, tissue preparation, and quantitative RNA and protein analysis for all 5 projects. DNA oligonucleotides will be synthesized using an Applied Biosciences 392 DNA synthesizer and purified by gel electrophoresis. These will be used for in situ hybridization protocols in projects 1,2, and 4. RNA and protein will be isolated from tissue samples used in each of the projects (human brain, experimental animals, or tissue culture material). Brains will be microdissected to allow analysis of discrete anatomical regions. RNA and protein separation will be carried out by RNAse protection assays and Western blots. Quantification will be by autoradiography and densitometry using an LKB Ultrascan Laser Densitometer. Providing these quantitative assays as a core service will enhance each project, and be cost effective compared to duplicate analytical setups in each laboratory. Particularly with regard to the human material, the core will allow for optimal use of scarce resources. Furthermore, in the course of these studies, the core will generate RNA and protein samples from carefully microdissected brain regions of clinically well characterized patients and young and old experimental animals. This will be an invaluable resource and will be of great use as these studies of aging and Alzheimer disease evolve.

该核心将为试剂合成，组织提供中心资源 制备和定量RNA和蛋白质分析 项目。 DNA寡核苷酸将使用施加的 Biosciences 392 DNA合成器，并通过凝胶电泳纯化。 这些将用于项目1,2的原位杂交协议 和4。将从每个中使用的组织样品中分离RNA和蛋白质 项目（人脑，实验动物或组织培养 材料）。 大脑将被微解释以允许分析离散 解剖区域。 RNA和蛋白质分离将通过 RNase保护测定法和蛋白质印迹。 量化将是 使用LKB Ultrascan激光器的放射自显影和光密度法 密度计。 提供这些定量测定作为核心服务将增强每个 项目，与重复的分析设置相比，具有成本效益 在每个实验室。 特别是关于人类物质的 核心将允许最佳利用稀缺资源。 此外，在 这些研究的过程，核心将产生RNA和蛋白质样品 从精心临床的精心分解的大脑区域 表征患者和年轻的和老式的实验动物。 这会 成为宝贵的资源，将非常有用，因为这些研究 衰老和阿尔茨海默氏病进化。