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

新的设备和软件将提高佛罗里达州立大学测序设施的生产力和扩展能力，这是一个核心设施，为来自生物科学不同学科的33多个用户的研究需求提供服务。应用生物系统公司（ABI）的PRISM 377自动DNA测序仪将补充目前的仪器，一个10岁的ABI 373 A型。新的测序仪将不仅允许在更短的时间内以更高的准确性对更大数量的样品进行测序和处理，而且它还将允许DNA片段（例如，RAPD，AFLP和微卫星）分析，一个操作，目前无法与我们目前的机器配置。另外的测序相关仪器和软件如下：（1）设备和软件形式的升级，其将扩展新旧ABI DNA测序仪的能力;（2）Mastercycler梯度热循环仪，其将允许需要不同引物退火温度的DNA模板的同时循环测序;（3）一台Macintosh G4 PowerPC服务器，用于存储DNA序列和片段输出，以及允许多个用户远程访问数据和序列分析程序的软件;（4）Macintosh和PC格式的许可证，最多可供12名用户同时使用Sequencher序列分析程序;（5）彩色打印机和CD读/写ROM，以支持FSU测序设施和Macintosh G4服务器中的计算机;（6）一个数码凝胶成像和记录系统，该系统比我们现有的系统更敏感和更通用，并有助于检测低-产生DNA样本用于ABI 377测序仪的后续分析。该设备将积极影响由七个主要用户，26个额外的教师用户，他们的研究生将核心设施目前的测序能力增加一倍以上，并首次允许自动DNA片段分析.目前的测序项目解决了进化生物学和系统学、结构生物学、细胞生物学、微生物生态学、肌肉生理学、神经生物学和遗传学中的问题。每个凝胶更多的DNA样品，每个样品更多的核苷酸读取，更精确的凝胶跟踪，更灵敏的样品检测，和更短的电泳时间）;（2）更快和更有效的数据处理、存储和检索手段;（3）更灵活、有效和用户友好的序列分析平台;（4）执行新研究方法的能力（例如，自动化DNA片段分析）;和（5）PCR扩增产物和测序模板的更好的定量和质量评估。FSU致力于提升其研究使命，并将在不久的将来聘请大量新的生物学教师。仪器升级是必要的，以吸引高质量的教师申请人，为研究生提供与国家的最先进的技术工作的经验，并适应目前的需求和预期的增加自动DNA测序相关的研究。

Abstract LevitanNew 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.