AB3500xL DNA Analyzer for OHSU Core Facility

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DESCRIPTION (provided by applicant): Funds for an Applied Biosystems / Life Technology 3500xL (24 capillary) DNA Analyzer are requested. It will become the principle instrument for the DNA sequencing and fragment analysis provided by the DNA Services Core Facility (://www.ohsu.edu/xd/research/research-cores/dna- analysis/). This well-established core facility provides capillary electrophoresis sequencing services to ~100 principle investigators in 23 OHSU departments, center and institutes, and 12 other non-OHSU Portland area departments. Long-read Sanger sequencing is a basic technique required by all labs performing modern, hypothesis-driven molecular biology. Massively parallel whole genome research is expected to increase the need for follow-up Sanger sequencing in the near term. We will run ~30,000 samples this year. With the requested instrument, we will be able to double that productivity. A few, of the more than 200 basic and translational NIH funded projects we support, are "Accelerating target identification and validation in leukemia" in the Brian Druker lab in the Knight Cancer Institute; "Treatment of Distal Tyrosine pathway Disorders" in the Markus Grompe lab in the Oregon Stem Cell Center; and "Genetic and molecular dissection of hair-cell function" in the Teresa Nicolson lab in the School of Medicine, Otolaryngology Research Department. Sequencing by our core for projects in Molecular Microbiology & Immunology, Infectious Disease, Ophthalmology, Cell Biology & Development and Nephrology departments in the School of Medicine, the Center for Research on Occupational & Environmental Toxicology (CROET) and the Biology Department at Portland State University round out our major user group; accounting for >55% of our current usage. The PI for this grant designed and implemented the databases and website used by the core for receiving work-requests, tracking work and performing billing and reporting functions. The technical staff, besides Dr. Keller, consists of one full-time technician with several years of automated DNA sequencing experience and one-half time technician. The current, 16 capillary 3130xL sequencer will be used for shorter, faster PCR sequencing runs; and for backup during maintenance and repair downtime. The requested 24 capillary instrument will be used for samples requiring the long reads (800-1000 bases) and fragment analysis. The requested instrumentation will significantly increase our data throughput and genomic analysis productivity.