IGEM:UC Berkeley/2006/Ligation
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< IGEM:UC Berkeley/2006(Difference between revisions)
Current revision (16:38, 26 June 2006) (view source) |
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| + | '''Basic Ligation''' | ||
#Create Ligation Mixture: | #Create Ligation Mixture: | ||
#*5.5 uL water | #*5.5 uL water | ||
| Line 8: | Line 9: | ||
#*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: | + | '''Scaled-Up Ligations:''' |
*Use this for the construction of libraries. | *Use this for the construction of libraries. | ||
| - | + | #Create Ligation mixture: | |
| - | + | #*69uL H2O | |
| - | # 69uL H2O | + | #*10uL Ligase buffer |
| - | # 10uL Ligase buffer | + | #*10 uL backbone |
| - | # 10 uL backbone | + | #*10uL insert |
| - | # 10uL insert | + | #*1uL Ligase |
| - | # 1uL Ligase | + | #Incubate at room temperature for 60 minutes. |
| - | #Incubate at room temperature for | + | |
Current revision
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.
