IGEM:UC Berkeley/2006/Ligation

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Current revision (15:38, 26 June 2006) (view source)
 
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1. Create Ligation Mixture:<br>
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'''Basic Ligation'''
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  * 5.5 uL water
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#Create Ligation Mixture:
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  * 1 uL ligation buffer
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#*5.5 uL water
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  * 0.5 uL ligase
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#*1 uL ligation buffer
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  * 1 uL backbone (pAC997)
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#*0.5 uL ligase
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  * 2 uL insert <br>
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#*1 uL backbone (pAC997)
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#*2 uL insert  
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#Let ligate at room temp for 30 minutes <br>
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#*Note: For the control, add 2 uL of water instead of the insert
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2. Let ligate at room temp for 30 minutes <br>
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'''Scaled-Up Ligations:'''
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Note: For the control, add 2 uL of water instead of the insert
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*Use this for the construction of libraries.
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#Create Ligation mixture:
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#*69uL H2O
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#*10uL Ligase buffer
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#*10 uL backbone
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#*10uL insert
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#*1uL Ligase
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#Incubate at room temperature for 60 minutes.

Current revision

Basic Ligation

  1. Create Ligation Mixture:
    • 5.5 uL water
    • 1 uL ligation buffer
    • 0.5 uL ligase
    • 1 uL backbone (pAC997)
    • 2 uL insert
  2. Let ligate at room temp for 30 minutes
    • Note: For the control, add 2 uL of water instead of the insert

Scaled-Up Ligations:

  • Use this for the construction of libraries.
  1. Create Ligation mixture:
    • 69uL H2O
    • 10uL Ligase buffer
    • 10 uL backbone
    • 10uL insert
    • 1uL Ligase
  2. Incubate at room temperature for 60 minutes.
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