This application seeks to expand and upgrade instrumentation for the CAMBI Nucleic Acid Facility. The current users of this facility are supported by at least 64 different NIH grants totaling over $7 million in annual direct costs. All users require molecular biological methods in their research which includes: the prevention of dental disease, analysis of viral diseases and viral assembly, metabolic diseases, fundamental research relevant to heart disease, neurological diseases and stroke, the molecular basis of aging, protein turnover and folding, the nature of malignant cells, and fundamental processes of gene control. The Facility synthesizes oligonucleotides, advises on oligo design and use, does automated DNA sequencing, and aids in the analysis of sequence data. An equipment package is requested for three purposes. The first is to upgrade oligonucleotide syntheses to make the operation more efficient, to lower oligo cost, and to provide services (including riboprobes and 40 nmol level synthesis) which are not practical with current instrumentation. The second is to upgrade the automated DNA sequencing by automating M13 template preparation and DNA sequencing reactions for more efficient operation and reduction in the cost of sample preparation. A further upgrade of DNA sequencer hardware and software is requested to increase the sequencing output by 2-3-fold per run, to facilitate heterozygote sequence analysis, and to facilitate assembly of large contigs (up to 50kb). Software is also requested to expand the capabilities of the existing sequencer for microsatellite- based quantitative genetic analysis and linkage studies. The third is to provide automated DNA and RNA purification from a variety of biological samples. A fee for service by the Facility covers supplies, service contracts, and funding for part time technical help as needed. The University provides support through the CAMBI bell and Advanced Molecular Biology and Immunology) group for a technician and a part-time accountant. This institutional support allows the Facility to provide services at below the cost of outside agencies and thus supplements NIH funded research. The Facility is available to the Buffalo area research community and is administered through CAMBI, group of research active faculty from 10 departments (Biochemical Pharmacology, Biological Sciences, Biochemistry, Biophysics, Chemical Engineering, Medicine, Microbiology, Oral Biology, Chemistry, and Pharmacology & Therapeutics) in 5 schools (Pharmacy, Natural Sciences & Mathematics, Medicine, Dentistry, and Engineering) at SUNY Buffalo. The elected CAMBI Steering Committee of 7 members plus 2 CAMBI Co-directors will serve as the internal advisory committee to review administration of the requested instrumentation. In the past 87.3% of the oligonucleotides synthesized by the Facility (for 64 laboratories) have been for NIH funded projects. In addition, 93.9% of the templates sequenced (for 42 laboratories) have been for NIH funded projects. At least 87% of the use of the instrumentation in this application will be for NIH-funded research.
