Transformation Protocol

From 2012.igem.org

Transformation Protocol:

Before you start: <p> • Prepare an ice bath. <p> • Prepare a 42 °C water bath. <p> • Pre-warm SOC buffer and plates at 37 °C. <p> • Autoclaved 1.5 mL Eppendorf tubes. <p> • 15 ml falcon tubes. <p> 1. Incubate both the plasmid preparation and the competent cells on ice for 15 minutes in separate 1.5 mL Eppendorf tubes. <p> 2. Combine 3 μL of plasmid prep. with approximately 500 μL of the competent cells preparation. <p> 3. Heat shock the mixture for 40 seconds at 42 °C, then immediately incubate it on ice for 2 minutes. <p> 4. Following this, add 1.0 mL of pre-warmed SOC buffer to your mixture, transfer to a falcon tube and incubate for 1 hour at 37 °C. <p> 5. Inoculate pre-warmed plates with 100 μL or 300 μL of the cell suspension and incubate overnight at 37 °C.