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Team:Calgary/Notebook/Protocols/pBAD
From 2011.igem.org
Notebook
Post-Regionals
Journal
- Defining Objectives
- Techniques Workshop
- Bioinformatics & qRT-PCR
- Sensory Element Fishing
- Electrochem - CPR Oxidation
- Electrochem - Reporter Gene
- Pseudomonas Tools
- Microalgae Tools
- NA Viability Assays
- May - July Group Journal
Protocols
Safety
Determining the Effectiveness of the pBAD-LacZ Construct
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.
