IGEM:IMPERIAL/2006/Protocols/I13207
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I13207 AiiA Test Construct Protocol
- Culture bacteria (DH5a) in LB medium, incubate overnight
- Transfer plasmids with desired part via electroporation
- Incubate overnight
- Transfer colonies into liquid medium (amp resistant, 2ml)
- Wait until optical density is 0.1
- Make 20mL of 1% arabinose LB solution
- Then make the following solutions:
| Concentration | How to set up solutions |
| 1.00% | 10mL 1.00% arabinose solution as made up above |
| 0.50% | 5 mL 1.00% and 5 mL Pure LB |
| 0.10% | 1 mL 1.00% and 9 mL Pure LB |
| 0.05% | 1 mL 0.05% and 9 mL Pure LB |
| 0.00% | 10 mL Pure LB |
- Transfer arabinose solutions into 5 different labeled tubes
- One well is filled with pure LB media in all experiments as a control
- Transfer 2 mL of each arabinose solution into separate clear tubes
- Add 200ul of DH5a culture into the tubes
- Assay with an automatically repeating protocol of absorbance measurements (600nm filter, 0.1 second absorbance through approximately 0.5cm of fluid)
- Fluorescence readings (488nm excitation filter, 525nm emission filter, 0.5 second, CW lamp energy 12902 units)
- Shake (1 mm, linear, normal speed)
- Time between measurements should be 20 minutes
- Time period for measurements should be ~5 h
- Perform 2 tests: one with 0.5ml of 0.5M ZnCl2 in the LB medium and one without
