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

摘要Levit新设备和软件将提高佛罗里达州立大学测序设施的生产率并扩展其能力，该设施是一项核心设施，为来自不同学科的33多名用户提供生物科学方面的研究需求。应用生物系统公司(ABI)的PRISM 377自动DNA测序仪将补充现有的仪器，即已有10年历史的ABI型号373A。新的测序仪不仅可以在更短的时间和更高的精度内对更多的样本进行测序和处理，而且还可以进行DNA片段(例如，RAPD、AFLP和微卫星)分析，这是目前我们的机器配置所无法完成的操作。其他与测序有关的仪器和软件如下：(1)以设备和软件的形式进行升级，以扩展旧的和新的ABI DNA测序仪的能力；(2)MasterCycler梯度热循环仪，允许同时对需要不同引物退火温度的DNA模板进行循环测序；(3)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12个用户可以同时访问Sequencher序列分析程序；(5)彩色打印机和CD读/写只读光盘，以支持FSU测序设备和Macintosh G4服务器中的计算机；(6)比我们目前的系统更灵敏和通用的数字凝胶成像和文档系统，它将有助于检测低产量DNA样本，以便随后在ABI 377测序仪上进行分析。该设备将对由7个主要用户、26个额外的教员用户及其研究生进行的非常多样化的研究产生积极影响，使核心设施的现有测序能力增加一倍以上，并首次允许进行自动化DNA片段分析。目前的测序项目涉及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。从这种仪器获得的主要好处是：(1)增加了以较低成本及时获取数据的能力(例如，每凝胶更多DNA样本、每样本更多核苷酸读取、更准确的凝胶跟踪、更灵敏的样本检测和更短的电泳时间)；(2)更快和更高效的数据处理、存储和检索方法；(3)更灵活、高效和用户友好的序列分析平台；(4)能够在现场执行新的研究方法(例如，自动化DNA片段分析)；以及(5)更好地量化和质量评估聚合酶链式反应扩增产物和测序模板。佛罗里达州立大学致力于提升其研究任务，并将在不久的将来招聘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者，为研究生提供使用最先进技术的经验，并适应目前的需求和自动化DNA测序相关研究的预期增长。

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.