A DNA Sequencing Facility for Interdisciplinary Biological Research

DNA序列测定对许多生物科学领域的研究具有重要的基础意义。包括功能生物化学、细胞和分子生物学、群体遗传学和分子系统学。该提案的目的是通过寻求资金建立DNA测序设施来简化亚利桑那州立大学的DNA测序。该设施的主要设备将是一台自动DNA测序仪，这是我们大学目前没有的，也是本提案所要求的。亚利桑那州立大学大约有20个研究小组在他们的研究中经常使用DNA测序，大量的研究生、本科生、博士后研究员、员工和教职员工将从拥有自动测序仪的DNA测序设施的建立中受益。所要求的测序仪利用红外激光二极管激发凝胶电泳大小分离的荧光标记DNA。检测荧光团的荧光，同时扫描四个测序通道，可以以高度经济高效的方式提供800个核苷酸或更多的DNA序列，准确率超过99%。配备所需设备，可同时对22个样品进行测序。除了提供30%的采购费用外，大学还将设立一个员工级别的职位，由个人管理DNA测序设施，并承担大部分涉及的人员成本。一位博士级别的研究科学家（已经在工作人员中）将监督该设施。象征性的用户费用（每个序列约5美元）将被收取，用于支付化学品、DNA测序仪服务合同的成本，以及技术人员工资的一小部分（约20%）。该设备将为获取和分析DNA序列信息提供一种高效、经济的机制。另一个好处是，DNA测序对大量在研究领域（例如，系统生物学和种群生物学）工作的研究生来说是可以接触到的，这些研究领域将受益于DNA分析，但对他们来说，DNA测序设备目前还不容易获得。此外，许多通常禁止使用放射性的本科项目可能包括DNA测序。获得自动DNA测序仪的另一个显著优势是，当DNA测序变得更有效时，学生将能够更方便地收集和分析序列信息，从而为其他重要领域的培训留下更多的空间。

DNA sequence determinations are of fundamental importance for studies in many areas of the biosciences. including functional biochemistry, cell and molecular biology, population genetics, and molecular systematics. The purpose of this proposal is to streamline DNA sequencing at Arizona State University by seeking funds to establish a DNA sequencing facility. The major piece of equipment for this facility will be an automated DNA sequencer, which currently is not available at our University and which is requested in this proposal. About 20 groups at Arizona State University routinely use DNA sequencing in their research, and a large number of graduate students, undergraduates, postdoctoral fellows, staff, and faculty will benefit from the establishment of a DNA sequencing facility with an automated sequencer. The sequencer requested utilizes an infrared laser diode to excite fluorophor-labeled DNA that is size-separated by gel electrophoresis. Fluorescence from the fluorophor is detected, and simultaneous scanning of four sequencing lanes over time can provide a DNA sequence of 800 nucleotides or more with over 99% accuracy in a highly cost-effective manner. With the equipment requested, 22 samples can be sequenced simultaneously. Apart from providing a 30% cost-share in purchasing this equipment, the University also will create a staff-level position for an individual to run the DNA sequencing facility and will bear most of the personnel cost involved. A Ph.D.-level research scientist (already on staff) will supervise the facility. Nominal user fees (about $5 per sequence) will be charged to cover the cost of chemicals, the DNA sequencer service contract, and a small part (about 20%) of the technician's salary. This facility will provide a highly effective and economic mechanism to obtain and analyze DNA sequence information. An additional benefit is that DNA sequencing becomes accessible to a large number of graduate students who work in research areas (for examp le, systematics and population biology) that would benefit from DNA analysis, but for whom DNA sequencing facilities currently are not readily available. Also, many undergraduate projects, which usually preclude use of radioactivity, could include DNA sequencing. Another significant advantage of acquisition of an automated DNA sequencer is that when DNA sequencing becomes more efficient, students will be able to gather and analyze sequence information more expediently, leaving more room for emphasis on training in other important areas.