The objective of the Molecular Biology Core unit will be to facilitate the timely utilization of molecular biology techniques by physicians and scientists in the Center. During the last four years, the core has evolved from three originally approved functions to the current six functions. The effort and performances of the core have won the praise of outside reviewers of this Center. Accordingly, we propose to continue all six services, each will be directed by an experienced investigator with specialized professional skill. Core functions will include: 1) To maintain and operative a share instrument facility, including an electroporator set, a Batch/Continuous Fermentator, an UVP gel documentation system, PCR machines, ultracentrifuges, and GCG Sequence Analysis Software Package. 2) To provide affordable and high quality sequencing service. Dr. Chan will provide this service with a semi-automated system (Applied Biosystem Inc. Model 373) at $10 per sequencing, which generally yields an average of 300-400 bp. 3) To provide assistance in applying the combinatorial library technique. The core will provide established libraries, assist new library construction, and provide hands-on technical assistance. 4) To provide assistance in applying transgenic and knock out methodologies. Dr. Tighe will assist investigators to initiate and perform transgenic mouse studies, and freeze and maintain frozen embryos of selected transgenic animals. 5) to provide assistance in conducting genetic studies of human autoimmune diseases. Dr. Nobori will assist investigators in performing linkage analyses and population genetics, positional cloning (i.e., reverse genetics), yeast artificial chromosome library screening and long-range mapping, and genome wide search for genetic factors associated with autoimmune diseases. 6) To provide technical assistance in basic molecular biology techniques and to organize the methodology seminars and user group meetings. Dr. Olee will teach basic techniques, and organize seminars and user group discussions on subjects of interest to investigators.
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