IGEM:UC Berkeley/2006/Ligation

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Line 12: Line 12:
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#69uL H2O
+
# 69uL H2O
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#10uL Ligase buffer
+
# 10uL Ligase buffer
-
#10 uL backbone
+
# 10 uL backbone
-
#10uL insert
+
# 10uL insert
-
#1uL Ligase
+
# 1uL Ligase
#Incubate at room temperature for 30 minutes.
#Incubate at room temperature for 30 minutes.

Revision as of 14:00, 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.


  1. 69uL H2O
  2. 10uL Ligase buffer
  3. 10 uL backbone
  4. 10uL insert
  5. 1uL Ligase
  6. Incubate at room temperature for 30 minutes.
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