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Revision as of 05:30, 11 September 2012 (view source) HSpedding (Talk | contribs) ← Older edit		Latest revision as of 02:18, 17 September 2012 (view source) DanWinter (Talk | contribs)
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-	Transformation Protocol:	+	{{:Team:Macquarie_Australia/Template/MQ12}}
-	Before you start:	+	== Transformation Protocol ==
-	• Prepare an ice bath.	+
-	• Prepare a 42 °C water bath.	+	Before you start:
		+	<br>
		+	• Prepare an ice bath.
-	• Pre-warm SOC buffer and plates at 37 °C.	+	• Prepare a 42 °C water bath.
-	• Autoclaved 1.5 mL Eppendorf tubes.	+	• Pre-warm SOC buffer and plates at 37 °C.
-	• 15 ml falcon tubes.	+	• 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.	+	1. Incubate both the plasmid preparation and the competent cells on ice for 15 minutes in separate 1.5 mL Eppendorf tubes.
-	3. Heat shock the mixture for 40 seconds at 42 °C, then immediately incubate it on ice for 2 minutes.	+	2. Combine 3 μL of plasmid prep. with approximately 500 μL of the competent cells preparation.
-	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.	+	3. Heat shock the mixture for 40 seconds at 42 °C, then immediately incubate it on ice for 2 minutes.
-	5. Inoculate pre-warmed plates with 100 μL or 300 μL of the cell suspension and incubate overnight at 37 °C.	+	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.

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Latest revision as of 02:18, 17 September 2012

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.