IGEM:UC Berkeley/2006/Ligation

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#Let ligate at room temp for 30 minutes <br>
#Let ligate at room temp for 30 minutes <br>
#*Note: For the control, add 2 uL of water instead of the insert
#*Note: For the control, add 2 uL of water instead of the insert
 +
 +
Scaled-Up Ligations:
 +
*Use this for the construction of libraries.
 +
 +
 +
#69uL H2O
 +
#10uL Ligase buffer
 +
#10 uL backbone
 +
#10uL insert
 +
#1uL Ligase
 +
#Incubate at room temperature for 30 minutes.

Revision as of 13:00, 26 June 2006

  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.


#69uL H2O
#10uL Ligase buffer
#10 uL backbone
#10uL insert
#1uL Ligase
  1. Incubate at room temperature for 30 minutes.
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