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Team:Macquarie Australia/Protocols/ligations
From 2012.igem.org
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<p>The mixtures were incubated for 10 minutes and then heat inactivated at 80°C for 20 minutes.</p> | <p>The mixtures were incubated for 10 minutes and then heat inactivated at 80°C for 20 minutes.</p> | ||
| - | <p>2µL of the ligation product was transformed into 50µL of competent <i>E. coli</i>. | + | <p>2µL of the ligation product was transformed into 50µL of competent <i>E. coli</i>, the transformants were then incubated overnight.</p> |
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Revision as of 04:42, 21 September 2012
Ligation Procedure
A reaction mixture was prepared containing,
| Element | Volume |
| Upstream Digestion part | 2 µL |
| Downstream Digestion part | 2 µL |
| Destination Plasmid | 2 µL |
| 10X T4 DNA ligase buffer | 2 µL |
| T4 DNA ligase | 1 µL |
| H2O | 11 µL |
The mixtures were incubated for 10 minutes and then heat inactivated at 80°C for 20 minutes.
2µL of the ligation product was transformed into 50µL of competent E. coli, the transformants were then incubated overnight.
