Team:Imperial College/Protocols

Below you will find a comprehensive list of all the protocols we used during our wet lab work. We spent some time toward the start of the project sourcing and updating effective protocols for B. subtilis manipulation explicitly, as other teams have previously had difficulty in this area.

These protocols worked for us, so any future teams looking to use subtilis could save quite a bit of time by looking here before diving into the literature!

1. Transformation Protocols for integration vectors in Bacillus subtilis

• Protocol 1
• Protocol 2

2. Transformation Protocol for E.coli

• Preparation of XL1-blue electrocompetent cells

3. PCR

• PCR

4. Calibration Curves

• Calibration Curves
• O.D ₆₀₀ vs cell number
• Fluorescence vs GFPmut3b

5. Promoter-RBS Testing Protocols

• Constitutive Promoters and RBS
• pHyperspank Promoter and RBS
• Light Inducible Promoters and RBS

6. Antibiotic Resistance Testing

• Chloramphenicol Resistance Assay