This proposal requests funding for a state of the art automated ABI 377 DNA sequencer to be operated by the Molecular Pharmacology Facility at the University of Washington. This facility has been in operation since 1989 and we are currently operating a first generation ABI DNA 370A sequencer at maximum capacity. We have now reached a critical stage where it is essential to upgrade our sequencing instrumentation in order to increase throughput, and provide the increased sensitivity and longer run lengths possible on the newer instrument. Increased sensitivity dyes have been developed which our filter based laser is unable to detect and we expect our current ABI instrument to become increasingly obsolete over the next year. The Pharmacology Facility was the first to purchase, through an NIH instrumentation grant, a fluorescent dye-based sequencing system and offer sequencing for u both within and outside our Department on a cost basis. Automated DNA sequencing has advanced rapidly over the past 10 years and has now largely replaced manual sequencing throughout the Media Center. A group of 16 principal investigators in 7 departments have accounted for about 75% of our DNA sequencing requests over the past year and they form the core group whose continued research productivity will be greatly assisted by the new instrumentation. The remaining 25% of last year's E sequencing in the Molecular Pharmacology Facility has been broadly distributed among another 54 investigators in 26 departments throughout the University of Washington. Funding of this proposal provide essential and cost effective technical services to a large group of highly productive laboratories funded by the NIH.
