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Team:Macquarie Australia/Protocols/Ligation
From 2013.igem.org
Ligation Procedure
A reaction mixture was prepared containing,
| Element | Volume |
|---|---|
| Upstream Digestion part | 2 µL |
| Downstream Digestion part | 2 µL |
| Destination Plasmid | 1 µL |
| 10X T4 DNA ligase buffer | 2 µL |
| T4 DNA ligase | 1 µL |
| H2O | 12 µL |
We incubated each ligation mix at 30°C for 30 minutes, followed by heat inactivation at 80°C for 20 minutes.
4 µL of the ligation product was transformed into 100 µL of competent E. coli, the transformants were then incubated for one hour.
