This core facility will provide equipment, reagents, and technical support required for the successful completion of the molecular biology components of the individual projeCts. The rationale of the core is that all of the projects require either molecular cloning or expression studies using cloned cDNAs. The molecular biology core is a single laboratory space, to be shared by all of the groups, which will alleviate duplication costs for equipment, supplies, and salaries. By nature of its central location in the Life Sciences Building, the core facility will enhance scientific interactions among project members. The core is also designed to eliminate potential technical barriers which may exist between labs of different disciplines. The molecular biology core will provide the following specific services critical for each proposed project: a) Propagation and maintenance of plasmid vectors: plasmids will be purified in the core using either CsCl gradient ultra centrifugation or commercial kits which utilize chromatography. The core will be responsible for transformation of the DNAs into bacteria, for maintaining frozen stocks of the recombinant bacteria, and for mailing DNAs to other colleagues in the field, upon request. b) Sequencing: Sequencing reactions will be performed by the research assistant using oligonucleotide primers made with the core Millipore synthesizer. A new automated sequencer in the Life Science building will be used as soon as it is up-and-running. A computer work station in core, containing DNA software, is available for storage and analysis of data. c) Preparation of commonly used solutions (electrophoresis buffers, bacterial media) and preparation of bacterial plates containing different antibiotics. d) Preparation of synthetic RNAs for expression studies. e) Purification of GST-fusion proteins. f) Maintenance of all equipment, including periodic cleaning of centrifuges, rotors, gel drier, lyophilizers.
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