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

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


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