This proposal requests funds for the purchase of a Perkin Elmer ABI DNA Sequencer 377 with upgrade to 96 lanes. The requested instrument package includes the automatic sequencing apparatus including gel electrophoresis unit, laser and fluorescence detection system as well as a computer (Macintosh) and sequence detection and analysis software. This system provides all the functionality required for the analysis of fluorescently labeled DNA sequences prepared by polymerase chain reaction (PCR). The instrument will be located in the State University of New York at Stony Brook (SUNY SB) DNA Sequencing Facility which is housed in the Department of Physiology and Biophysics. The sequencer will significantly promote the research objectives and laboratory productivity of fourteen distinguished scientists at SUNY SB who comprise the major users group. The sequencer will make available to these investigators the technology required for the rapid analysis of DNA sequences which is presently inadequately supported at SUNY SB and difficult to obtain from other organizations in the local area. Automatic DNA sequencing technology is far superior to manual sequencing, which most investigators have to rely upon in part or full, due to greatly increased throughput, read-length, accuracy, ease of use and the elimination of the requirement for radionucleotides. The automatic sequencer will not only facilitate currently active projects but will also open up new possibilities for investigators allowing them to tackle much larger sequencing projects than is currently feasible. The instrument will be operated and maintained by trained personnel at the SUNY SB DNA Sequencing Facility. The director of the facility will schedule daily operation and provide technical expertise for the users. An advisory committee will ensure access to the major users and other PHS- supported investigators and will be responsible for the long term operation of the instrument and the facility.
