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

采购用于研究和教育的实时定量 PCR 系统

• 批准号: 0116722
• 负责人: Phillip Danielson
• 金额: $ 10.04万
• 依托单位: University of Denver
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0116722&HistoricalAwards=false
• 关键词: Acquisition; Real; Time; Quantitative; PCR

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.

丹佛大学的Phillip Danielson博士获得了一笔资助，用于购买实时定量PCR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/研究生研究和教育的生产力和成本效益。将立即受益的具体研究项目包括国家科学基金会资助的研究：(1)生物体中负责疼痛和应激反应的大脑内分泌激素；(2)细胞色素P450毒素代谢酶——了解它对控制农业害虫和许多携带疾病的生物体至关重要；(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研究。直到最近，使用传统测定法测量基因表达意味着多次艰苦的优化、测试模板稀释和测定后操作。即使在那时，由于传统的基于端点的测量的不可预测的可变性，估计遗传信息的浓度往往是不准确的。这是因为在分析过程中，放大效率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。新一代实时定量PCR系统解决了这些问题。该系统实时监测反应动力学，使其能够以无与伦比的准确性，精度和速度定量最小组织样品中的DNA和RNA浓度。一个额外的好处是，PCR仪器也可以用于高速基因分型。丹佛大学已经具备的自动化液体处理能力将处理样品制备，通过最大限度地减少移液错误和交叉污染，确保运行到运行的一致性。除了丹佛大学教师的研究活动的好处之外，广泛的实验室和课堂导向的教育目标将在本科和研究生水平上得到推进。生物科学系提供的分子生物学理学学士学位和文学学士学位的核心是分子导向的实验课程，这些课程的好处将尤为明显。在更广泛的层面上，系里资助的面向高中学生的生物技术课程和促进中学一级动手科学教育的教师培训讲习班也将大大加强。考虑到这些项目针对的是城市和低收入学区的学生，而这些学生传统上在自然科学方面的代表性不足，高中推广工作的好处是不可估量的。简而言之，获得PCR系统将为中学、本科和研究生水平的教育提供显著和直接的好处，同时提供一种经济有效的方法来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。