This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This core facility aims to assist program and institution investigators in conducting consistent, high quality genome scale experiments. Based on the needs of Stem Cell COBRE and other investigators, the following services have been developed: Automated RNA and RNA purification: High throughput purification is performed using Autogen DNA and RNA purification systems in conjunction with an Autogen tissue homogenizer. Also, a Qiacube system provides versatility and speed for processing of smaller sample numbers. Quality assessment of nucleic acids: A Nanodrop spectrophotometer provides accurate quantitation and purity analyses. Quantitative RT-PCR: Two BioRad instruments are in use. The MyIQ system performs single-dye QPCR assay, while the IQ5 allows multiplexing of QPCR, providing expression analysis of up to 5 genes simultaneously per sample. The demand for multiplexing is expected to rise as COBRE investigators'projects increasingly focus on validating research findings in human patient material with limited availability. Microarray analysis: Since the number of MMCRI investigators performing transcriptome analyses does not warrant investment in microarray instrumentation, an active collaboration has been established with the COBRE-funded Microarray Facility at the University of Vermont. In addition to primary data, quality control/analysis is provided by a bioinformatician. The MMCRI Bioinformatics &Genomics Core coordinates investigator requests for microarray analyses, Affymetrix microarray and reagent purchases and billing to expedite this interaction. Furthermore, a fully equipped workstation is provided for users who prefer to process their own primary data, with access to both X-RAY and Genespring packages for primary data analysis, and Ingenuity for biological pathway analysis.
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