Team:EPF-Lausanne/Protocols/Transformation

From 2009.igem.org

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'''Procedure'''
'''Procedure'''
<br>
<br>
-
<br>1. Thaw cells on ice for 20min
+
:1. Thaw cells on ice for 20min
-
<br>2. Add DNA (ligation product) to the cells, mix.
+
:2. Add DNA (ligation product) to the cells, mix.
-
<br>3. Incubate 20min on ice
+
:3. Incubate 20min on ice
-
<br>4. Heat shock 45s @ 42°C
+
:4. Heat shock 45s @ 42°C
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<br>5. Incubate 20min on ice (optional)
+
:5. Incubate 20min on ice (optional)
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<br>6. Add 500 µl SOC and incubate 1h @ 37°C (shaking)
+
:6. Add 500 µl SOC and incubate 1h @ 37°C (shaking)
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<br>7. Spread over LB-agar plate
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:7. Spread over LB-agar plate
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<br>8. Incubate overnight @ 37°C
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:8. Incubate overnight @ 37°C
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</div><div CLASS="epfl09bouchon"></div>

Revision as of 07:11, 9 September 2009



Transformation



Material

  • 50 µl competent cells
  • LB-agar plates with corresponding antibiotics
  • Water bath at 42°C


Procedure

1. Thaw cells on ice for 20min
2. Add DNA (ligation product) to the cells, mix.
3. Incubate 20min on ice
4. Heat shock 45s @ 42°C
5. Incubate 20min on ice (optional)
6. Add 500 µl SOC and incubate 1h @ 37°C (shaking)
7. Spread over LB-agar plate
8. Incubate overnight @ 37°C


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