Full Title: High Throughput Algae Quantification using Quantitative Polymerase Chain Reaction for a Novel Biofuel Harvesting System by Dillon Flebbe
Advised by: Dr. Christopher Bachmann, Dr. Louie Wurch
Abstract: As concern over burning fossil fuels and its effect on the environment grows, so does the need for clean and renewable energy. Algae biofuels are a promising carbon-neutral source that could be adopted in place of liquid fossil fuels. The algae biofuel project at JMU is focused on the harvesting aspect of algae biofuel production, with the goal of improving energy return on investment and economic feasibility, which has yet to be proven. Analyzing energy return and economic feasibility requires the number of cells being processed to be known. Currently the project is using spectrophotometry and a hemocytometer to obtain cell counts. These methods however have inadequacies. Spectrophotometry is the quick and easy method, but the results vary based on the amount of chlorophyll present leading to inaccurate measurements. Currently counting cells with a hemocytometer is considered the best option because it is believed to give accurate counts, but it is extremely slow and relies on the operator to count the cells. Preliminary data indicates a large standard deviation (+/- 1.73 million cells/ ml) associated with this method, likely due to human error. QPCR is a high throughput method of quantifying based on replicating DNA and fluorescent dye binding to the newly created DNA products. While significantly more expensive due to specialized machinery and consumable reagents, because it is based on DNA the cell counts it produces are accurate. A qPCR protocol was developed for the Nannochloropsis algae and utilized as an alternative to the hemocytometer. Based on preliminary data, there appears to be room for improvement which qPCR can exploit. Using qPCR to quickly and accurately count cells can aid in the research of algae based biofuels and possibly lead to the fuels’ implementation.
Post time: Oct-03-2017