Difference between revisions of "IGEM:UC Berkeley/2006/Ligation"

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'''Basic Ligation'''
 
#Create Ligation Mixture:
 
#Create Ligation Mixture:
 
#*5.5 uL water
 
#*5.5 uL water
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'''Scaled-Up Ligations:'''
 
'''Scaled-Up Ligations:'''
 
*Use this for the construction of libraries.
 
*Use this for the construction of libraries.
# 69uL H2O
+
#Create Ligation mixture:
# 10uL Ligase buffer
+
#*69uL H2O
# 10 uL backbone
+
#*10uL Ligase buffer
# 10uL insert
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#*10 uL backbone
# 1uL Ligase
+
#*10uL insert
#Incubate at room temperature for 30 minutes.
+
#*1uL Ligase
 +
#Incubate at room temperature for 60 minutes.

Latest revision as of 13:38, 26 June 2006

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.