IGEM:UC Berkeley/2006/Ligation: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
Line 12: | Line 12: | ||
# 69uL H2O | |||
# 10uL Ligase buffer | |||
# 10 uL backbone | |||
# 10uL insert | |||
# 1uL Ligase | |||
#Incubate at room temperature for 30 minutes. | #Incubate at room temperature for 30 minutes. |
Revision as of 11:00, 26 June 2006
- 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.
- 69uL H2O
- 10uL Ligase buffer
- 10 uL backbone
- 10uL insert
- 1uL Ligase
- Incubate at room temperature for 30 minutes.