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Biofuel Notebook

6/28/12

We ran a test of the Ethanol Assay to determine its range of effectiveness.

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7/3/12

We ordered materials for the ethanol assay and set up experiments to grow cells anaerobically to see if ethanol could be detected.

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7/9/12

We started an aerobic and an anaerobic liquid culture of E. coli cells so that we could measure ethanol secreted the next day.

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7/10/12

We used a plate reader to measure the ethanol concentrations but found both anaerobic and aerobic cultures produced elevated ethanol levels. This could have been because the cells grew too quickly and many were forced into anaerobic conditions in the aerobic culture. The OD (optical density) level of the aerobic culture was elevated.

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7/13/12

We decided to try a few new ways to decrease aerobic ethanol yields and decided to try the experiment in flasks in a shaking incubator.

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7/17/12

We made additional MOPS buffer and then set our aerobic culture experiment to incubate in a shaker overnight.

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7/18/12

We ran an ethanol assay on our aerobic cultures. We found that the cells had overgrown in the aerobic culture tubes and were likely producing ethanol due to forced anaerobic conditions.

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7/19/12

We decided to do the ethanol assay again, this time in aerated flasks, in order to decrease the chance of anaerobic growth. The cultures were put in MOPS media again.

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7/20/12

Since the cultures once again showed high ethanol levels after the ethanol assay, we prepared a timepoint assay for the next day. We put e. coli in flasks with 20 ml MOPS and then took OD measurements at 3 hour timepoints to determine when ethanol production was low. We took timepoints at 1.30, 3.15, 6.30, and 22.15 from the start of the culture, and used two different colonies in two different flasks. The results are below.

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