IGEM:UC Berkeley/2006/Ligation: Difference between revisions
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#*10uL insert | #*10uL insert | ||
#*1uL Ligase | #*1uL Ligase | ||
#Incubate at room temperature for | #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.