This application requests funds for an Applied Biosystems 3100 capillary DNA sequencer. The sequencer will be used for high volume DNA sequencing and DNA fragment analysis. The sequencer will be operated by the Molecular and Cellular Biology Core of the Oregon Regional Primate Research Center (ORPRC) an institute of Oregon Health Sciences University (OHSU) and serve the entire, newly-created, OHSU West Campus located 12 miles from the main campus. Needs of major users, who are supported by NIH R0l and Center Grants, will include analysis of T-cell function that will generate 19,000 sequences annually, analyses of T cell haplotypes that will generate 12,000 sizing reaction annually, analysis of subtracted cDNA libraries that will generate 2,500 sequencing reactions annually, analysis of microarrays and differential display that will generate 2,000 reactions annually. The sequencer will also support the newly established OHSU microarray facility by sequencing 5,000 cDNA clones annually and the function of the MCB core itself by 3,000 sequencing reactions annually. Minor users will require an additional 5,000 sequencing reactions annually. Because of this demand a high throughput machine is requested. The 3100 is a highly automated 16 capillary machine capable of running 7 days a week, 24 hours per day, from 9 to 16 runs per capillary per day. To process samples efficiently, the following accessories are requested: (1) An Applied Biosystems model 9700 PCR machine with twin 384 sample blocks to prepare samples rapidly enough to keep up with the capacity of the 3100. (2) The Applied Biosystems Comparative Sequencing Software to allow rapid assembly and identification of obtained DNA sequences. (3) The Applied Biosystems Gene Mapper software package for DNA fragment analysis. This software combined with the DNA sequencer will allow users to rapidly obtain and process the thousands of reactions necessary for their research programs. An internal advisory Committee consisting of the Principal Investigator and three major users will monitor use of the sequencer and develop guidelines for shared use of the instrument. The Principal Investigator in consultation with the advisory committee will establish procedures and oversee general operation of the sequencer. The Principal Investigator has headed the Molecular and Cellular Biology Core at the ORPRC for the previous seven years and has 6 years experience with automated sequencing. A full time, Master?s level technician with 6 years experience with automated sequencers will tend to day to day operation of the sequencer. Currently the only DNA sequencer on the OHSU West Campus is an outdated ABI373 that this machine will replace. Institutional commitment to this application includes providing full service contracts, all needed renovations, back up power (generator and UPS), software upgrades and salary support for the technical staff.
