NIDDK BIOTECHNOLOGY CENTER AT THE UNIVERSITY OF FLORIDA

DESCRIPTION (Abstract of the application) As the Human Genome Project exponentially increases the amount of DNA sequence information available to researchers, the paradigm of analyzing a single gene effect in a biological system has shifted to a global systems analysis. New high-throughput technologies have created unparalleled opportunities to study the mechanism of diseases, monitor the disease progression and evaluate effective therapies. Gene expression profiling is a critical tool to accomplish these ambitious goals. Because these technologies require expensive infrastructure and extensive technical expertise, they are not yet available to most biomedical researchers. The goal of this proposal is to expand our cDNA microarray facility and to make it available to a large number of NIDDK funded investigators. Our Specific Aims are: 1) To establish a high throughput cDNA microarray facility and perform microarray analysis for NIDDK-funded investigators. We are requesting resources for the following purposes: a) To increase the capacity of our microarray facility through purchase of additional equipment, computers and software, and cDNA clones. b) To develop software for automatic sample and data tracking and storage, statistical evaluation of arrays (quality control, reproducibility), innovative web-based data mining and statistical analysis. c) To perform microarray analysis for center participants. We also plan to distribute arrays at cost for other investigators. 2) To establish a training/education program for microarray technologies. The center will assume the responsibility to educate and train the participants on applications of microarrays, experimental design and protocols, data management (navigation of our secure web site & retrieval of their own array data), data analysis and data interpretation. Both workshop and individualized training for theory and hands-on bench work will be provided. 3) To improve microarray assay sensitivity through our R&D effort. We will evaluate: a) new labeling fluorophores (near infrared fluorophores from Li-Cor: IRD700 and IRD800), b) new labeling schemes using the rolling circle amplification (RCA) method, and c) alternative methods for attaching the target DNA to the surface of the solid support.