An Automated DNA Sequencer and Related Equipment for Multidisciplinary Biological Studies at Florida State University

Florida State University, Tallahassee FL

Investigators

Abstract

Abstract Levitan New equipment and software will enhance the productivity and extend the capabilities of the Florida State University Sequencing Facility, a core facility that services the research needs of 33-plus users from diverse disciplines in the biological sciences. An Applied Biosystems (ABI) PRISM 377 Automated DNA Sequencer will supplement the current instrument, a ten-year old ABI Model 373A. The new sequencer will not only permit a larger number of samples to be sequenced and processed in less time and with greater accuracy, but it will also permit DNA fragment (e.g., RAPD, AFLP, and microsatellite) analysis, an operation that is presently unavailable with the configuration of our current machine. Additional sequencing-related instrumentation and software is as follows: (1) upgrades in the form of equipment and software that will extend the capabilities of both old and new ABI DNA sequencers; (2) a Mastercycler gradient thermal cycler that will permit simultaneous cycle-sequencing of DNA templates requiring different primer annealing temperatures; (3) a Macintosh G4 PowerPC server to store DNA sequence and fragment output, and software that will permit remote access to data and sequence-analysis programs by multiple users; (4) Macintosh- and PC-format licenses for simultaneous access to the Sequencher Sequence Analysis Program by up to 12 users; (5) color printers and CD read/write ROMs to support computers in the FSU Sequencing Facility and the Macintosh G4 server; (6) a digital gel-imaging and documentation system that is more sensitive and versatile than our current system and one that will facilitate the detection of low-yield DNA samples for subsequent analysis on the ABI 377 Sequencer. This equipment will positively impact a remarkably diverse body of research conducted by seven major users, 26 additional faculty users, and their graduate students by more than doubling the present sequencing capacity of the core facility and for the first time permitting automated DNA fragment analysis. Current sequencing projects address questions in evolutionary biology and systematics, structural biology, cellular biology, microbial ecology, muscle physiology, neurobiology and genetics. The major benefits to be gained from this instrumentation are: (1) an increase in the timely acquisition of data at lower cost (e.g., more DNA samples per gel, more nucleotides read per sample, more accurate gel-tracking, more sensitive sample detection, and shorter electrophoresis times); (2) a faster and more efficient means of data handling, storage and retrieval; (3) a more flexible, efficient and user-friendly sequence-analysis platform; (4) the capability to perform new research methodologies (e.g., automated DNA fragment analysis) on-site; and (5) better quantification and quality-assessment of PCR-amplification products and sequencing templates. FSU is committed to upscale its research mission and a substantial number of new biology faculty will be hired in the near future. The instrumentation upgrade is necessary to attract high-quality faculty applicants, to provide graduate students the experience of working with state-of-the-art technology, and to accommodate the present demand and anticipated increase in automated DNA sequencing-related research.