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Team:Macquarie Australia/Protocols/Ligation
From 2013.igem.org
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| + | <b>Prepare</b> the following reaction mixture | ||
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<thead><tr><th>Element </th><th>Volume</th></tr></thead> | <thead><tr><th>Element </th><th>Volume</th></tr></thead> | ||
<tbody><tr><td>Upstream Digestion part </td><td>2 µL</td></tr> | <tbody><tr><td>Upstream Digestion part </td><td>2 µL</td></tr> | ||
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| + | Incubate each ligation mix at 30°C for 30 minutes, followed by heat inactivation at 80°C for 20 minutes. | ||
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| + | transform 4 µL of the ligation product into 100 µL of competent E. coli, then incubate the transformants for one hour. | ||
| - | + | [[File:LigationQLD.jpg|500px|thumb|center|Ligation]] | |
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Latest revision as of 07:18, 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.
