CORE--DNA SEQUENCING

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The DNA sequencing core facility has been an important component of the NEOMAMDC since 1993, and has significantly enhanced the research infrastructure of the Center, and accelerated progress for Arthritis researchers. Although the scope of molecular biology procedures used by laboratories associated with the Arthritis Center is quite broad and are project-specific most laboratories of the NEOMAMDC have benefitted and will continue to benefit from a core facility dedicated to DNA sequence determination. The rationale for this is that 1) DNA sequencing is required at some point in any molecular biology oriented experimental approach, and 2) it is not always cost effective for each laboratory to set up individual protocols, purchase equipment an train the necessary personnel. 3) DNA sequence analysis is labor and time intensive, but does not require specific knowledge of the large project. 4) The on site sequencing facility enhances the overall productivity of Arthritis Center research laboratories by performing this task more efficiently and with greater precision than would be possible on an individual basis 5) The core facility frees time for investigators to engage in more creative pursuits. The primary objective of the core is to assist those investigators already engaged in molecular biology protocols. In order to be practical, it does not serve to perform any molecular biology technique other than DNA sequence analysis. Investigators are responsible for preparation of samples for sequencing in advance of submission. Protocols for samples preparation are provided by the sequencing core in order to assure that the core facility will be efficiently utilize and to assure success. In general, investigators are responsible for ligation of DNA fragments into the polylinker (cloning site) of sequencing vectors commonly in use. The core facility is then responsible for performing the sequencing reactions, running the automated sequencing instrument and recording the sequence as a file that can be used for further analysis by the investigator. The second objective of the core is to serve as repository for genomic DNA from members of families afflicted with OA showing familial inheritance, and from patients with OA disease subsets. DNA from these individuals is made available to other investigators on a collaborative basis. Additionally, the core functions to screen this DNA bank for mutations in the type II collagen gene.