CORE--CELL /MOLECULAR /MICROSCOPY

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The molecular genetics and proteomics core serves as a resource to provide expertise and assistance in the analysis of gene and protein expression to projects by Van Eyk and Tomaselli. The emphasis of the core will be on the preparation and utilization of mRNA and protein for the performance and analysis of microarray and protein expression studies. Activities of the core facility will be performed in several locations on the campus of the Johns Hopkins School of Medicine including laboratories in the Division of Cardiology, the Sidney Kimmel Comprehensive Cancer Care Microarray Facility and the Center for Computational Medicine and Biology centered in the Department of Biomedical Engineering. The services provided by this Core include: mRNA preparation and qualification for kinetic real time PCR (RT-PCR) and microarray experiments; access to canine EST libraries; spotting and hybridization to microarrays, assistance with analysis of microarray data; access to CAGE, a database repository for microarray experiments; primers for RT-PCR; canine tissue qualified antibodies and protocols for their use in Western blotting, immunohistochemical and immunocytochemical applications; cytosolic, sarcolemmal, sarcoplasmic reticulum, ERK1/2 and connexin subproteome preparation and analysis (including the performance of 2D gel electrophoresis (2DE), 2D liquid chromatography (2DLC), differential imaging gel electrophoresis (DICE) and mass spectrometry); and a canine heart-specific protein database for peptide mass fingerprinting (PMF). In addition to the provision of technical expertise to the projects, the core will generate significant cost savings by serving as a centralized resource for molecular biological and protein chemical analysis of the canine heart.