Funding is requested for the purchase of a Li-Cor 4000LS automated deoxyribonucleic acid (DNA) sequencer. This equipment is to be used by no fewer than 5 separate project groups involving a diverse range of research disciplines at the College of William and Mary. Four of five principal investigators are faculty at the Virginia Institute of Marine Science (VIMS), the School of Marine Science campus of the College located in Gloucester Point, VA. Research project groups at VIMS are conducting studies on 1) the population genetics, biogeography and taxonomy of selected marine organisms with emphasis on pelagic fishes, 2) the biology of the phylum Haplosporidia and the phylogenetic relationship of these protists to other eukaryotes, 3) the phylogenetics and systematics of the Clitellata including leeches, oligochaetes and branchiobdellidans, 4) the association of major histocompatibility complex II gene alleles in trout with immune responsiveness, and 5) the origin and maintenance of biodiversity in marine invertebrate taxa. In addition to the VIMS project groups, studies in the Department of Biology of the College involve screening genes that can be used as dorsal-ventral markers in Xenopus to examine the tissue interactions leading to the determination of the dorsal-ventral axis in vertebrates. The collective need for an automated sequencer has been directed by 3 principal criteria: 1) data throughput and efficiency, 2) radiation safety and 3) centralization. The purchase of the Li-Cor 4000LS automated sequencer offers considerable advantages to throughput and efficiency over manual methods currently available at the College of William and Mary. These advantages include increased base identification accuracy, increased read-length from a single primer, and greatly reduced time investment. The use of an automated sequencer will also eliminate radioactive waste and will reduce the risk of environmental exposure. Currently, sequencing reactions at the College ar e run in the presence of 35S labeling. The use of radioactive material entails the risk of exposure both to the experimenter and to those working in the same facilities and severely curtails the opportunities for students, especially undergraduates, to gain experience with DNA sequencing. In addition, waste disposal is a problem. The single radioactive waste disposal site available for eastern Virginia will close on December 31, 1995 and a substitute facility is not scheduled to be available until the year 2000. In the interim, all radioactive waste generated at the College must be stored on site. The laser-detection of infrared excitation in an automated sequencer does not involve the use of radioactive labeling methods and thus circumvents impending limitations imposed by accumulating radioactive waste and experimenter exposure. Moreover, the non-radioactive detection method makes the technology freely available and safe for teaching purposes at any level. Centralization of all sequencing operations will enhance existing interactions among all project groups.
