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Team:Macquarie Australia/Protocols/Ligation
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Revision as of 07:10, 13 September 2013
Ligation Procedure
Prepare the following reaction mixture
| 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 |
Incubate each ligation mix at 30°C for 30 minutes, followed by heat inactivation at 80°C for 20 minutes.
transform 4 µL of the ligation product into 100 µL of competent E. coli, then incubate the transformants for one hour.
