Difference between revisions of "Team:Kyoto/Experiments"
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<h2>Ligation</h2> | <h2>Ligation</h2> | ||
| − | Ligation was performed according to Ligation high Ver.2’s protocol. | + | <p>Ligation was performed according to Ligation high Ver.2’s protocol. |
1. unfreeze competent cells on hands. put them on ice. | 1. unfreeze competent cells on hands. put them on ice. | ||
| Line 145: | Line 145: | ||
3. keep them in heating block 45 seconds, then on ice for 30 minutes. | 3. keep them in heating block 45 seconds, then on ice for 30 minutes. | ||
4. add 40~200ul SOC liquid culture medium, then incubate it under shaking culture for 1hour 37℃. | 4. add 40~200ul SOC liquid culture medium, then incubate it under shaking culture for 1hour 37℃. | ||
| − | 5. seed it onto LB+antibiotics +/- culture. incubate it for 24 hour at 37℃. | + | 5. seed it onto LB+antibiotics +/- culture. incubate it for 24 hour at 37℃.</p> |
Revision as of 09:16, 12 October 2016
Protocols
Ligation
Ligation was performed according to Ligation high Ver.2’s protocol. 1. unfreeze competent cells on hands. put them on ice. 2. add DNA sample (1~20ul) to competent cells. leave them on ice for 30 minutes 3. keep them in heating block 45 seconds, then on ice for 30 minutes. 4. add 40~200ul SOC liquid culture medium, then incubate it under shaking culture for 1hour 37℃. 5. seed it onto LB+antibiotics +/- culture. incubate it for 24 hour at 37℃.
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