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Monday 7/5
Note: Today was another Institute Holiday.
Tuesday 7/6
- Hydrolyzed linseed oil. The reaction solution will be characterized on Wednesday.
Wednesday 7/7
- Attempted to make [http://partsregistry.org/Part:BBa_V1012 V1012] (strain CP919, KanR, envZ deficient) electrocompetent:
- Centrifuged overnight 3mL LB-Kan culture at 4C, 16000rcf for 10min.
- Discarded supernatant. Resuspended pellet in 1.5mL ice-cold water. Centrifuged again.
- Repeated with 0.75mL ice-cold water.
- Resuspended pellet in 1.5mL ice-cold 10% glycerol.
- Flash-froze 100uL aliquots in dry ice/ethanol.
- Transformed light/lysis ligation product from last week into [http://partsregistry.org/Part:BBa_V1012 V1012] via electroporation. (Voltage: 2.5V; time constant: 4.5)
- Plated 100uL on LB-Tet agar plate & inoculated a 5mL LB-Tet overnight culture.
- Inoculated 5mL LB-Kan culture of purely [http://partsregistry.org/Part:BBa_V1012 V1012] in case we need to retry the competence procedure.
- Tested pigment production of cells containing [http://partsregistry.org/Part:BBa_K274100 K274100] in various solutions.
- Tested solutions of LB, LB without yeast extract added, 80% glycerol, 100% glycerol, and glycerol with different concentrations of agar.
- Since the brick is under an arabinose-inducible promoter, two versions of each solution were made - one containing arabinose, and one without arabinose.
- Pigment production was subjectively measured every hour for five hours.
Thursday 7/8
- The V1012 transformation appears to have worked - the plate is covered in 1000+ colonies.
- Made freezer stock from parallel liquid culture.
- Miniprepped the DNA for further modification and for sequencing.
Friday 7/9
Weekend 7/10-11
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