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Team:Arizona State/notebook
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<p> </p> | <p> </p> | ||
<p><strong>Procedures</strong> <br> | <p><strong>Procedures</strong> <br> | ||
| - | Gel making/running. | + | Gel making/running: Gels were all 1% agarose gels using safe white dye. Al were ran for 1 hr. at 95 V. |
| - | + | <p> | |
| - | Enyzme digestions. | + | Enyzme digestions/ Ligations: Our team had a bit of a learning curve with digestions and ligations. We tried several different methods for digestions to ensure we had full digestion, but most the most common was the procedure found in the <a href="http://openwetware.org/wiki/Haynes:Assembly101">Haynes Lab Procedures</a> |
Ligations. | Ligations. | ||
| - | + | <p> </p> | |
| + | <p><strong>Resting Cell Assay</strong> <p> | ||
| + | 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. </p> | ||
| + | <p> </p> | ||
| + | <p><strong>Quantification</strong> <p> | ||
| + | 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. | ||
</p> | </p> | ||
Revision as of 16:44, 17 October 2014
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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 Enyzme digestions/ Ligations: Our team had a bit of a learning curve with digestions and ligations. We tried several different methods for digestions to ensure we had full digestion, but most the most common was the procedure found in the Haynes Lab Procedures Ligations.
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.
Daily entries from open wet ware?
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