Difference between revisions of "Team:Kyoto/Experiments"
| Line 138: | Line 138: | ||
<h1>Protocols</h1> | <h1>Protocols</h1> | ||
<h2>Ligation</h2> | <h2>Ligation</h2> | ||
| + | |||
| + | <p>Ligation was performed according to Ligation high Ver.2’s protocol.</p> | ||
| + | |||
| + | <p>1. unfreeze competent cells on hands. put them on ice.<br> | ||
| + | 2. add DNA sample (1~20ul) to competent cells. leave them on ice for 30 minutes<br> | ||
| + | 3. keep them in heating block 45 seconds, then on ice for 30 minutes.<br> | ||
| + | 4. add 40~200ul SOC liquid culture medium, then incubate it under shaking culture for 1hour 37℃.<br> | ||
| + | 5. seed it onto LB+antibiotics +/- culture. incubate it for 24 hour at 37℃.</p> | ||
| + | |||
| + | <h2>PCR</h2> | ||
<p>Ligation was performed according to Ligation high Ver.2’s protocol.</p> | <p>Ligation was performed according to Ligation high Ver.2’s protocol.</p> | ||
Revision as of 09:24, 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℃.
PCR
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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