MOLECULAR BIOLOGY

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Molecular Biology Module has supported investigators of the Vision COBRE in their research programs primarily through DNA purification from tissues and PCR-based genotyping. The module also supports investigators through maintenance of equipment used for gene expression analysis by real-time PCR, and Western blots. Dr. Ash is the director of this module and he oversees its day to day operation. Dr Ash is qualified to supervise this module based on his extensive experience in real-time PCR, Western blot analysis, DNA purification, and PCR genotyping. Dr Ash has worked with genetically modified mice since 1994. During this 14 year period, Dr Ash has maintained approximately 30 lines of genetically modified mice. His lab currently genotypes 5 to 10,000 samples per year. Dr. Carr is the assistant director. Dr Carr has extensive experience in animal genotyping, as well as analysis by Real-Time PCR and ELISA. Operations of the Molecular Biology Module are conduced by a full time research assistant Fatemeh Shariati. Projects are initiated by a service request. Following completion of service Mrs. Shariati completes a report describing the completion of the job and all results. The report will contain a gel picture documenting accurate PCR reactions, and an assessment of positive and negative results. All reports are approved by the module director before they are returned to the users. If the results are not satisfactory, Mrs. Shariati will repeat the analysis. The module is operating at a rate of 10,000 reactions per year with and accuracy rate of 80 to 90%.