IGEM:UC Berkeley/2006/Ligation: Difference between revisions
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'''Basic Ligation''' | |||
#Create Ligation Mixture: | #Create Ligation Mixture: | ||
#*5.5 uL water | #*5.5 uL water | ||
| Line 7: | Line 8: | ||
#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. | |||
#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
- Create Ligation Mixture:
- 5.5 uL water
- 1 uL ligation buffer
- 0.5 uL ligase
- 1 uL backbone (pAC997)
- 2 uL insert
- 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.
- Create Ligation mixture:
- 69uL H2O
- 10uL Ligase buffer
- 10 uL backbone
- 10uL insert
- 1uL Ligase
- Incubate at room temperature for 60 minutes.
