Acquisition of a Real-Time Quantitative PCR System for Research and Education

奖项摘要丹佛大学的菲利普·丹尼尔森博士获得了一笔赠款，用于资助购买实时定量聚合酶链式反应系统。基于成熟的激光荧光和DNA扩增技术，该系统将显著提高分子生物学本科生/研究生研究和教育的生产率和成本效益。将立即受益的具体研究计划包括：(1)大脑中负责生物疼痛和应激反应的内分泌激素；(2)细胞色素P450毒素代谢酶--了解这种酶对于控制农业害虫和许多携带疾病的生物至关重要；(3)丹佛大学与丹佛动物园和丹佛科学与自然博物馆合作开展的濒危和受威胁物种的遗传多样性。直到最近，使用传统分析方法测量基因表达还意味着无数轮费力的优化、测试模板稀释和分析后处理。即使在那时，由于传统的基于终点的测量的不可预测的变异性，估计的遗传信息浓度往往是不准确的。这是因为在检测过程中，扩增效率上的小偏差会在被测量的最终产物的数量上产生很大的差异。在新一代实时荧光定量聚合酶链式反应系统中找到了解决这些问题的方法。该系统实时监控反应动力学，使得能够以无与伦比的准确性、精密度和速度对最小的组织样本中的DNA和RNA浓度进行定量。一个额外的好处是，该聚合酶链式反应仪器也可以用于高速基因分型。丹佛大学已有的自动化液体处理能力将处理样品制备，通过将进样误差和交叉污染降至最低，确保连续运行的一致性。除了丹佛大学教职员工的研究活动受益外，本科生和研究生都将推进一系列以实验室和课堂为导向的教育目标。生物科学系新开设的分子生物学学士和文学士学位的核心是以分子为导向的实验室课程，这些课程的好处将特别明显。在更广泛的层面上，为高中生提供的部门赞助的生物技术课程和促进中学科学实践教育的教师培训讲习班也将得到极大加强。鉴于这些计划的对象是城市和低收入学区的学生，而这些学生在自然科学领域的代表性一直较低，高中推广工作的好处是不可估量的。简而言之，购买聚合酶链式反应系统将为中学、本科和研究生教育提供显著和直接的好处，同时提供一种经济高效的手段，满足由国家科学基金会资助的生命科学研究人员日益增长的DNA分析需求。

Award Abstract A grant has been awarded to Dr. Phillip Danielson at the University ofDenver to fund the purchase of a real-time quantitative PCR System. Based on well-established laser fluorescence and DNA amplification technology, this system will significantly increase the productivity and cost effectiveness of undergraduate/graduate research and education in molecular biology. Specific research programs that will benefit immediately include National Science Foundation-funded studies on: (1) endocrine hormones in the brain that are responsible for pain and stress responses in living organisms; (2) cytochrome P450 toxin-metabolizing enzymes - an understanding of which is critical to the control of agricultural pests and many disease-carrying organisms and; (3) the genetic diversity of endangered and threatened species which the University of Denver conducts in collaboration with the Denver Zoological Gardens and the Denver Museum of Science and Nature. Until recently, the measurement of gene expression using traditional assays has meant numerous rounds of laborious optimization, test template dilutions and post assay manipulations. Even then, the estimated concentration of a genetic message was often inaccurate owing to the unpredictable variability of traditional endpoint-based measurements. This is because small biases in amplification efficiency over the course of an assay would produce large differences in the amount of final product being measured. A solution to these problems was found in the new generation of real-time quantitative PCR Systems. The system monitors reaction kinetics in real-time making it possible to quantitate DNA and RNA concentrations in the smallest of tissue samples with unparalleled accuracy, precision and speed. An added benefit is that the PCR instrument can also be used for high-speed genotyping. The automated liquid handling capabilities already in place at the University of Denver will handle sample preparation to ensure run-to-run consistency by minimizing pipetting errors and crossover contamination. Beyond the benefit to the research activities of faculty at the University of Denver, a broad range of laboratory and classroom-oriented educational goals will be advanced at both the undergraduate and graduate levels. Benefits will be particularly evident in the molecular-oriented laboratory courses that are at the heart of the new Bachelor of Science and Bachelor of Arts degrees in Molecular Biology offered by the Department of Biological Sciences. On a broader level, department-sponsored biotechnology classes offered to high school students and teacher-training workshops that promote hands-on science education at the secondary school level will also be greatly enhanced. The benefit to high school outreach efforts is immeasurable given that these programs target students in urban and low-income school districts who have traditionally been underrepresented in the natural sciences. In short, acquisition of the PCR System will provide significant and immediate benefits to education at the secondary, undergraduate and graduate levels, while providing a cost-efficient means of satisfying the growing DNA analysis needs of life-science researchers funded by the National Science Foundation.