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.
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