CORE--BRAIN MICRODISSECTION, RNA AND PROTEIN ANALYSIS

核心——脑显微切割、RNA和蛋白质分析

• 批准号: 6098488
• 负责人: BRADLEY T. HYMAN
• 金额: $ 14.43万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6098488
• 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 所有 5 种药物的制备、RNA 和蛋白质定量分析 项目。 DNA寡核苷酸将使用Applied公司合成 Biosciences 392 DNA 合成仪并通过凝胶电泳纯化。 这些将用于项目 1,2, 4. RNA 和蛋白质将从每个中使用的组织样本中分离出来。 项目（人脑、实验动物或组织培养 材料）。 大脑将被显微解剖，以便分析离散的 解剖区域。 RNA 和蛋白质分离将通过 RNAse 保护测定和蛋白质印迹。 量化将通过 使用 LKB Ultrascan 激光进行放射自显影和光密度测定 密度计。 提供这些定量检测作为核心服务将增强每一项服务 项目，并且与重复的分析设置相比具有成本效益 在每个实验室。 特别是在人体物质方面， 核心将允许优化利用稀缺资源。 此外，在 在这些研究过程中，核心将生成 RNA 和蛋白质样本 来自临床良好的仔细显微解剖的大脑区域 描述了患者和年轻和年老实验动物的特征。 这将 是一种宝贵的资源，并且将在这些研究中发挥很大的作用 衰老和阿尔茨海默病不断发展。