The human genome project over the past five years has provided a wealth of basic genetic information. At the same time the computational resources and equipment for analysis of genetic polymorphisms and genetic sequences has advanced to the stage where complex genetic disorders, such as type I diabetes are amenable to analysis. Sequence based information has already revolutionized study of """"""""simple"""""""" Mendelian disorders and now is likely to have a similar impact of diseases such as type I diabetes. In addition, type I diabetes as an autoimmune disorder is a field where pathogenesis is critically dependent upon genetic rearrangement and diversity generation within T cell receptors and immunoglobulin genes. The ability to effectively study type I diabetes is therefore critically dependent upon the feasibility of sequencing multiple genes, multiple alleles of multiple genes, multiple T cell receptors, multiple target autoantigens, and the genotyping of families with type I diabetes and genotyping of animal crosses. The current proposal is from a group of principal investigators at the University of Colorado Health Sciences Center who have a pressing need to acquire the technology for high throughput and automated sequencing and genotyping as a primary research tool. A request is made based upon acquiring the instrumentation of a Qiagen Biorobot 9600 and GenAMP PCR System 9700 for automated plasmid preparation and polymerase chain reaction (PCR), combined with the automated sequencing and genotyping capabilities of an ABI 377-01 prism sequinator. There is at present, at the University of Colorado Health Sciences Center, only six older model sequinators and no Biorobotic system for a campus with more than 900 faculty. These machines are developed to diverse areas of research including Cancer research, immunology National Jewish) or pharmacology. The proposed user group is centered on the Barbara Davis Center for Childhood Diabetes and includes 6 of its principal investigators, including its Executive Director (G. Eisenbarth) and Research Director (J. Hutton). This user group, with its expanding demands for sequence information and genomic typing, has exceeded the capacity of a single older model, 36 lane ABI sequinator and lacks access to the capabilities of a Biorobot to automated large numbers of sequencing and genotyping reactions. Affiliate members of the group are involved in active collaboration with the Barbara Davis Center and are located nearby in the department of Cellular and Structural Biology, Microbiology, and Biostatistics and Preventative Medicine. NIH funding for the user group totals approximately $2.75 million per year. An advisory group comprises of two members from the BDC and two members from the Health Sciences Center's departments of Cellular and Structural Biology will provide a balanced management policy for all major users. User fees will be shared to fund the management and maintenance of the equipment, calculations being based on the accumulated usage time on the instruments. Unforeseen operation costs not covered by these fees will be absorbed by the BDC with funds provided to the Executive Director, George Eisenbarth, who is the Principal Investigator on this proposal.
