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The estimation of the amount or copy number of a specific gene is difficult and dependent on a number of parameters. The accuracy and precision of the measurement relies on the quality of the target nucleic acids as well as the robustness of the measurement protocol. We are focusing on real-time quantitative PCR (polymerase chain reaction) measurements, which utilize intercalating dyes or specific fluorescent-labeled probes for monitoring the amplification of the target DNAs. We are working with a plasmid-based system for measurement and calibration of the fluorescent signal with respect to different kinds of fluorescent probes. A pilot study with the Bioanalytical Working Group of the CCQM (committee of the International Bureau of Weights and Measures) is in progress. We seek to understand the importance of various parameters in the overall accuracy of the measurements. Biotech grain constitutes a large proportion of the US corn and soybean harvest. Detection and quantitative estimates of biotech grain content in whole grain and processed food samples has become an important issue for international trade in these commodity crops. Quantitative DNA technologies including quantitative real-time PCR offer the current best prospect for such measurements. Determination of the amount and quality of DNA, isolated from materials that will be subject to testing, are critical parameters. Method validation, improvements in DNA isolation, and accurate quantitation of total DNA are key issues. A current emphasis is on developing primary methods for quantitating total genomic DNA from plants for the purpose of certifying Standard Reference Materials. The performance of fluorescent DNA-binding dyes with various plant genomic DNAs is under investigation. Biocatalytic systems Enzymes are increasingly important for large- and small-scale synthesis in the pharmaceutical and fine chemical industries. Research is focused on several enzyme systems of known or potential value. Microbial enzymes are cloned, mutated and expressed in quantities for studies of structure, kinetics and other properties of native and modified enzymes. | ||
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