CORE--DE PEDIATRIC COBRE: CELL SCIENCE CORE

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A Cell Science Core is critical to the development of all five of the projects in this proposed Center. The Core brings together all of the cell and molecular biological resources available within the department, and provides access to additional services available at the University of Delaware and the Delaware Biotechnology inst. Within the Core, a preparative facility will be established to procure human tissues for DNA, RNA, and protein isolation. Primary human cell cultures will also be established to support investigators' projects. The tissues and cells will be obtained through the Clinical Services Core (Core B) to ensure patient confidentiality and compliance with federal, state and institutional regulations. The preparative core will create EBV-immortalized lymphoblasts, and primary fibroblasts from skin and other tissues. An Analytical Facility will be created, consisting of a Cell Biology Resource, a Protein Biochemistry Resource, and a Molecular Biology Sub-Core. The Cell Biology Resource will centralize use and availability of instrumentation and provide operational and experimental expertise in cell science. The resources will include Confocal microscopy, fluorescence and visible spectroscopy, and FACS. Through an agreement with the Delaware Biotechnology Inst., multiphoton confocal microscopy, trasmission and scanning electron microscopy, and atomic force microscopy will also be available. The Protein Biochemistry Resource will bring together equipment and expertise for protein expression, purification, and characterization. 2D gel analysis systems will be developed, and an agreement with the Univ. of Delaware will provide extended capabilities to identify proteins by MALDI /TOF/TOF and a variety of altemative mass spectrometric techniques. The Molecular Biology Sub-Core will provide access to and expertise in a range of modem molecular techniques for DNA sequencing and genetic analysis. These techniques include pyrosequencing, mutation analysis by denaturing high performance liquid chromatography, and real-time PCR. The creation of a unified Cell Science Core wilt provide a unique facility to provide each investigator access to multiple modern experimental approaches. The experienced investigators in the Core will interact proactively with the project investigators to enhance project development.