- 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
- Use this for the construction of libraries.
#69uL H2O #10uL Ligase buffer #10 uL backbone #10uL insert #1uL Ligase
- Incubate at room temperature for 30 minutes.