Dilute the overnight culture to 1/100th in 5 mL CaCl2
- At OD600 ≈ 1, take 5 Eppendorf tubes
1. Negative control tube – P1 lysate + 750 uL of recipient cells
2. 5 uL tube of P1 lysate + 750 uL of recipient cells
3. 10 uL tube of P1 lysate + 750 uL of recipient cells
4. 15 uL tube of P1 lysate + 750 uL of recipient cells
5. 20 uL tube of P1 lysate + 750 uL of recipient cells
Incubate for 20 min at 37°C WITHOUT stirring