A MICROBIAL BREATHALYZER: DESIGN OF A COLORIMETRIC ASSAY FOR THE DETECTION AND QUANTIFICATION OF ETHANOL PRODUCTION IN MICROBESPublic
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This project aims to address the issues of efficiency with the design of a three step assay that allows for the rapid screening of large numbers of microbial colonies for ability to produce ethanol from a variety of feedstocks. First, the microbes are grown on agar plates that are adjusted to have a basic pH and contain bromothymol blue, a pH indicator. Ethanol production also results in carbon dioxide production, changing the pH of the agar around the colonies to acidic and the color to shades of yellow and orange. These colonies are then treated with pyridinium chlorochromate, an oxidizer that changes color in the presence of primary and secondary alcohols. This rate of color change is measured spectrophotometrically, and is proportional to the concentration of ethanol in each culture.
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