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

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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.