Transformation Protocol

Revision as of 05:30, 11 September 2012 by HSpedding (Talk | contribs)

Transformation Protocol:

Before you start: • Prepare an ice bath.

• Prepare a 42 °C water bath.

• Pre-warm SOC buffer and plates at 37 °C.

• Autoclaved 1.5 mL Eppendorf tubes.

• 15 ml falcon tubes.

1. Incubate both the plasmid preparation and the competent cells on ice for 15 minutes in separate 1.5 mL Eppendorf tubes.

2. Combine 3 μL of plasmid prep. with approximately 500 μL of the competent cells preparation.

3. Heat shock the mixture for 40 seconds at 42 °C, then immediately incubate it on ice for 2 minutes.

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.

5. Inoculate pre-warmed plates with 100 μL or 300 μL of the cell suspension and incubate overnight at 37 °C.