DNA Sequencer (Applied Biosystems 3100)

DESCRIPTION: (provided by applicant): 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.