application): The mission of the Molecular Core, as conceived at the inception of this grant in 1993, was to facilitate the introduction and use of molecular genetic methods by a full range of potential DERC investigators at Yale. A preliminary assessment of investigators? needs was carried out by questionnaire and on this basis we assembled resources and personnel so as to provide: a) services, b) training, c) consultation and d) equipment and facilities dedicated to support a number of essential molecular techniques. The structure of the core was organized around three specific areas: 1) RNA analysis (RNase protection, nuclear run-off, and t 1/2 determination), 2) RNA localization (in situ hybridization) and 3) gene expression (in bacteria, cell lines and transgenic mice). Advice and consultation were to be offered for a number of related methodologies, including cDNA and genomic library construction, reporter gene assays, site-directed mutagenesis and PCR-based techniques. We specifically did not include resources such as DNA or protein sequencing and oligonucleotide or peptide synthesis, which are offered by other core facilities at Yale. A reagent/supplies storage function was likewise determined to be redundant; for example, sources for both restriction enzymes and for cell culture media and additives are already available in house.
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