Transformation Protocol

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(Difference between revisions)

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.

@@ Line 1: / Line 1: @@
-Transformation Protocol:
+{{:Team:Macquarie_Australia/Template/MQ12}}
-<p>
-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>
-.	Incubate both the plasmid preparation and the competent cells on ice for 15 minutes in separate 1.5 mL Eppendorf tubes.
+== Transformation Protocol ==
-<p>
-.	Combine 3 μL of plasmid prep. with approximately 500 μL of the competent cells preparation.
+Before you start:
-<p>
+<br>
-.	Heat shock the mixture for 40 seconds at 42 °C, then immediately incubate it on ice for 2 minutes.
+• Prepare an ice bath.
-<p>
-.	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.
+• Prepare a 42 °C water bath.
-<p>
-.	Inoculate pre-warmed plates with 100 μL or 300 μL of the cell suspension and incubate overnight at 37 °C.
+• Pre-warm SOC buffer and plates at 37 °C.
+• Autoclaved 1.5 mL Eppendorf tubes.
+• 15 ml falcon tubes.
+. Incubate both the plasmid preparation and the competent cells on ice for 15 minutes in separate 1.5 mL Eppendorf tubes.
+. Combine 3 μL of plasmid prep. with approximately 500 μL of the competent cells preparation.
+. Heat shock the mixture for 40 seconds at 42 °C, then immediately incubate it on ice for 2 minutes.
+. 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.
+. Inoculate pre-warmed plates with 100 μL or 300 μL of the cell suspension and incubate overnight at 37 °C.