Team:Arizona State/notebook

Notebook

Project Phases

June: Project planning and set up

July-October: Synthesis of Fatty acid producing plasmids, such as the TesA plasmid

September - October: Preparation and testing of the ethanol producing parts

October: Preparation and testing of the wax esterase plasmid

Procedures
Gel making/running: Gels were all 1% agarose gels using safe white dye. Al were ran for 1 hr. at 95 V.

Enyzme digestions/ Ligations: Our team had a bit of a learning curve with digestions and ligations. We tried several different methods to ensure they would work, but the most common was the procedure found in the Haynes Lab Procedures

Resting Cell Assay

To test the productivity of different plasmids, resting cell assays were run. All assays were run in a shaker incubator at 32 degrees C and 200 RPM. A phosphate buffered solution was used to suspend the growth of the cultures.

Quantification

A GC with a DB5 column was used to quantify data from all assays. For testing ethanol production, the method used had an initial temp. of 60 degrees C, and ramped by 10 degrees C until it reached 250 degrees C, which was then held for 3 minutes.