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Team:Macquarie Australia/Protocols/Transformations
From 2013.igem.org
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'''1.''' Incubate both the plasmid preparation and the competent cells on ice for 15 minutes in separate 1.5 mL Eppendorf tubes. | '''1.''' Incubate both the plasmid preparation and the competent cells on ice for 15 minutes in separate 1.5 mL Eppendorf tubes. | ||
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Latest revision as of 05:55, 29 August 2013
Transformation Protocol
Prep
• 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.
Methods
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.
