Team:Calgary/Notebook/Protocols/pBAD
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Determining the Effectiveness of the pBAD-LacZ Construct
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Protocol for the Experiment to Determine the Effectiveness of the pBAD-LacZ Construct
- Cells carrying the I0500_B0034_I732005 construct were cultured in 5mL LB broth (Kan) overnight.
- This was then subcultured in 15 mL LB until log phase (OD of ~0.6).
- Transfer 9 1.5 mL samples of the culture into 2mL tubes, pellet them and remove supernatant.
- To these add 1.9 mL of a solution of 2% arabinose and 3mM CPRG.
- Some of this solution is used to blank the victor plate reader.
- After mixing the solution with the cells, the cells are resuspended.
- At each time point one of the tubes was removed, pelleted, and 200uL of the supernatant taken for a reading.
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