This application requests partial support for the development of an Automated DNA Sequencing Facility. Seven independent laboratories in the Departments of Biochemistry, Botany, Genetics, and Zoology at the University of Washington, for whom DNA sequencing represents a major research focus (major users), and an additional seven laboratories in these Departments with a significant but less extensive need for DNA sequencing (minor users), are participating in this proposal. The impetus for this application derives from the realization by these individuals that existing methods for manual sequencing of DNA are slower, more work-intensive, and more expensive than are automated methods that employ fluorescent compounds to track DNA fragments. All 14 faculty members involved in this application have concluded that the need to rely on manual methods for DNA sequencing is having a distinctly negative impact on the progress of their research. The 14 projects that comprise this application are all funded by one or more research grants from the National Science Foundation, National Institutes of Health, and other granting agencies. The major projects involve: an analysis of genes required for cell migrations during oogenesis in Drosophila (C. Berg); the structure and function of the mammalian metaxin gene family (P. Bornstein); a molecular analysis of marine plant systems (R. Cattolico); large-scale sequencing of the MHC class II region in birds (S. Edwards); comparative sequence analysis of the two largest subunits of nuclear RNA polymerase II from different taxa (B. Hall); regulatory analyses of muscle creatine kinase genes (S. Hauschka); and hormonal regulation of insect development (L. Riddiford). The minor projects involve: characterization of the revolta gene of Arabidopsis thaliana (L. Comai); identification of components of the microtubule organizing center in S. cerevisiae (T. Davis); genetic control of the yeast cell cycle (L. Hartwell); regulation of mesodermal patterning in Xenopus and zebrafish (D. Kimelman); origin of polarity in Drosophila development (H. Ruohola-Baker); heterochromatic genes and position effects in Drosophila (B. akimoto); and evolution of a regulatory protein and its target gene in yeast (E.Young). We request the purchase of a model 377 ABI Prism Automated DNA Sequencer with associated computer software, and a GeneAmp PCR System 9600. These instruments will be housed in space provided by the Department of Biochemistry. The facility will be run on a fee-for-service basis (cost center). It is anticipated that 80-90% of the capacity of the facility will be utilized by the participating laboratories and that the remaining capacity will be made available to investigators at the University of Washington on a first-come, first-served basis.
