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Line 2: |
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| <div id="kyoto-notebook"> | | <div id="kyoto-notebook"> |
| <!-- 警告:ここより上を消したら怒る --> | | <!-- 警告:ここより上を消したら怒る --> |
- | <div>
| |
- |
| |
- | <a name="0731" class="kyoto-jump"></a>
| |
- | <h3>7/31</h3>
| |
- |
| |
- | <h4>Restriction Enzyme Digestion</h4>
| |
- | <span class="kyoto-author">Nakamura and Matsumoto</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th colspan=2>Sample/(µL)</th>
| |
- | <th>EcoR1/(µL)</th>
| |
- | <th>Xba1/(µL)</th>
| |
- | <th>Spe1/(µL)</th>
| |
- | <th>Pst1/(µL)</th>
| |
- | <th colspan=2>Buffer/(µL)</th>
| |
- | <th>BSA/(µL)</th>
| |
- | <th>MilliQ/(µL)</th>
| |
- | <th>Total/(µL)</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>3/24 PCR(dddD) cloning product</td>
| |
- | <td>3.2</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>H</td>
| |
- | <td>3</td>
| |
- | <td>-</td>
| |
- | <td>21.8</td>
| |
- | <td>30</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>7/18 RBS30@</td>
| |
- | <td>18.5</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>H</td>
| |
- | <td>3</td>
| |
- | <td>-</td>
| |
- | <td>6.5</td>
| |
- | <td>30</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>7/18 RBS30 control@</td>
| |
- | <td>6.2</td>
| |
- | <td>-</td>
| |
- | <td>-</td>
| |
- | <td>-</td>
| |
- | <td>-</td>
| |
- | <td>H</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>2.8</td>
| |
- | <td>10</td>
| |
- | </tr>
| |
- | </table>
| |
- | </div>
| |
- |
| |
- | <div>
| |
- |
| |
- | <a name="0801" class="kyoto-jump"></a>
| |
- | <h3>8/1</h3>
| |
- |
| |
- | <h4>Gel Extraction</h4>
| |
- | <span class="kyoto-author">Murata and Yasuda</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Lane</th>
| |
- | <th>Sample</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>1</td>
| |
- | <td>1kbp ladder</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>2</td>
| |
- | <td>3/24 PCR(dddD) cloning product E,S</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>3</td>
| |
- | <td>3/24 PCR(dddD) cloning product E,S</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>4</td>
| |
- | <td>Blank</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>5</td>
| |
- | <td>7/18 RBS30 E,S@</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>6</td>
| |
- | <td>7/18 RBS30 E,S@</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>7</td>
| |
- | <td>Blank</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>8</td>
| |
- | <td>1kbp ladder</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>9</td>
| |
- | <td>3/24 PCR(dddD) cloning product control</td>
| |
- | <tr>
| |
- | <td>10</td>
| |
- | <td>7/18 RBS30 control@</td>
| |
- | </tr>
| |
- | </table>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Sample</th>
| |
- | <th>Concentration/(µg/mL)</th>
| |
- | <th>260/280</th>
| |
- | <th>260/230</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>PSB1C3</td>
| |
- | <td>13.3</td>
| |
- | <td>1.90</td>
| |
- | <td>-</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>dddD</td>
| |
- | <td>10.8</td>
| |
- | <td>1.64</td>
| |
- | <td>0.15</td>
| |
- | </tr>
| |
- | </table>
| |
- |
| |
- | <h4>Ligation</h4>
| |
- | <span class="kyoto-author">Yasuda</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th colspan=2>Vector/(µL)</th>
| |
- | <th colspan=2>Insert/(µL)</th>
| |
- | <th colspan=2>Buffer/(µL)</th>
| |
- | <th colspan=2>Ligase/(µL)</th>
| |
- | <th>MilliQ/(µL)</th>
| |
- | <th>Total/(µL)</tr>
| |
- | <tr>
| |
- | <td>8/1 PSB1C3 gel extraction product 2047bp</td>
| |
- | <td>1.5</td>
| |
- | <td>8/1 dddD gel extraction product 2545bp</td>
| |
- | <td>6.9</td>
| |
- | <td>10xT4Ligase</td>
| |
- | <td>2</td>
| |
- | <td>T4Ligase</td>
| |
- | <td>1</td>
| |
- | <td>8.6</td>
| |
- | <td>20</td>
| |
- | </tr>
| |
- | </table>
| |
- |
| |
- | <h4>Transformation</h4>
| |
- | <span class="kyoto-author">Murata</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th colspan=2>Sample/(µL)</th>
| |
- | <th colspan=2>CompetentCells/(µL)</th>
| |
- | <th>Total/(µL)</th>
| |
- | <th>Medium</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>PSB1C3-dddD</td>
| |
- | <td>3</td>
| |
- | <td>コンピ名</td>
| |
- | <td>30</td>
| |
- | <td>33</td>
| |
- | <td>SOC</td>
| |
- | </tr>
| |
- | </table>
| |
- |
| |
- | </div>
| |
- |
| |
- | <div>
| |
- |
| |
- | <a name="0802" class="kyoto-jump"></a>
| |
- | <h3>8/2</h3>
| |
- |
| |
- | <h4>PCR Cloning</h4>
| |
- | <span class="kyoto-author">Murata</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th>@</th>
| |
- | <th>2x KAPA HiFi Hotstart Ready Mix/(µL)</th>
| |
- | <th>F Pre-dddD cloning 62/(µL)</th>
| |
- | <th>Rv Spe1-dddD cloning 68/(µL)</th>
| |
- | <th>MilliQ/(µL)</th>
| |
- | <th>Total/(µL)</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>@</td>
| |
- | <td>12.5</td>
| |
- | <td>0.75</td>
| |
- | <td>0.75</td>
| |
- | <td>11</td>
| |
- | <td>25</td>
| |
- | </tr>
| |
- | </table>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Predenature</th>
| |
- | <th>Denature</th>
| |
- | <th>Annealing</th>
| |
- | <th>Extension</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>95@</td>
| |
- | <td>98@</td>
| |
- | <td>65@</td>
| |
- | <td>72@</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>5min</td>
| |
- | <td>20sec</td>
| |
- | <td>15sec</td>
| |
- | <td>50sec</td>
| |
- | </tr>
| |
- | </table>
| |
- | <p>Denature-Annealing-Extension 25cycle</p>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Number</th>
| |
- | <th>Sample</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>1</td>
| |
- | <td>pSB1C3-dddD(1)</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>2</td>
| |
- | <td>pSB1C3-dddD(2)</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>3</td>
| |
- | <td>pSB1C3-dddD(3)</td>
| |
- | </tr>
| |
- | </table>
| |
- |
| |
- | <h4>DNA Purification</h4>
| |
- | <span class="kyoto-author">Murata</span>
| |
- | <p>@</p>
| |
- |
| |
- | <h4>Colony PCR</h4>
| |
- | <span class="kyoto-author">Honda,Nakamura and Matsumoto</span>
| |
- | <p>Master Mix</p>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Quick Tag/(µL)</th>
| |
- | <th>VF2/(µL)</th>
| |
- | <th>VR/(µL)</th>
| |
- | <th>MilliQ/(µL)</th>
| |
- | <th>Total/(µL)</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>15</td>
| |
- | <td>0.6</td>
| |
- | <td>0.6</td>
| |
- | <td>13.8</td>
| |
- | <td>30</td>
| |
- | </tr>
| |
- | </table>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Predenature</th>
| |
- | <th>Denature</th>
| |
- | <th>Annealing</th>
| |
- | <th>Extension</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>94@</td>
| |
- | <td>94@</td>
| |
- | <td>55@</td>
| |
- | <td>68@</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>2min</td>
| |
- | <td>30sec</td>
| |
- | <td>30sec</td>
| |
- | <td>24sec</td>
| |
- | </tr>
| |
- | </table>
| |
- | <p>Denature-Annealing-Extension 30cycle</p>
| |
- | <p>☆</p>
| |
- |
| |
- | <h4>Concentration Measurement</h4>
| |
- | <span class="kyoto-author">Nakamura and Matsumoto</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Sample</th>
| |
- | <th>Concentration/(µg/mL)</th>
| |
- | <th>260/280</th>
| |
- | <th>260/230</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>@</td>
| |
- | <td>245</td>
| |
- | <td>1.59</td>
| |
- | <td>2.17</td>
| |
- | </tr>
| |
- | </table>
| |
- |
| |
- | <h4>Restriction Enzyme Digestion</h4>
| |
- | <span class="kyoto-author">名前</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th colspan=2>Sample/(µL)</th>
| |
- | <th>EcoR1/(µL)</th>
| |
- | <th>Xba1/(µL)</th>
| |
- | <th>Spe1/(µL)</th>
| |
- | <th>Pst1/(µL)</th>
| |
- | <th colspan=2>Buffer/(µL)</th>
| |
- | <th>BSA/(µL)</th>
| |
- | <th>MilliQ/(µL)</th>
| |
- | <th>Total/(µL)</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>8/2 dddD cloning product@</td>
| |
- | <td>8.2</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>H</td>
| |
- | <td>3</td>
| |
- | <td>-</td>
| |
- | <td>16.8</td>
| |
- | <td>30</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>8/2 dddD cloning control@</td>
| |
- | <td>0.41</td>
| |
- | <td>-</td>
| |
- | <td>-</td>
| |
- | <td>-</td>
| |
- | <td>-</td>
| |
- | <td>H</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>8.6</td>
| |
- | <td>10</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>7/18 RBS30@</td>
| |
- | <td>12.7</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>H</td>
| |
- | <td>3</td>
| |
- | <td>-</td>
| |
- | <td>12.3</td>
| |
- | <td>30</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>7/18 RBS30 control@</td>
| |
- | <td>0.04</td>
| |
- | <td>-</td>
| |
- | <td>-</td>
| |
- | <td>-</td>
| |
- | <td>-</td>
| |
- | <td>H</td>
| |
- | <td>1</td>
| |
- | <td>-</td>
| |
- | <td>8.4</td>
| |
- | <td>10</td>
| |
- | </tr>
| |
- | </table>
| |
- |
| |
- | </div>
| |
- |
| |
- | <div>
| |
- |
| |
- | <a name="0803" class="kyoto-jump"></a>
| |
- | <h3>8/3</h3>
| |
- |
| |
- | <h4>@</h4>
| |
- | <span class="kyoto-author">Murata</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Lane</th>
| |
- | <th>Sample</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>1</td>
| |
- | <td>pSB1C3-dddD(1)</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>2</td>
| |
- | <td>pSB1C3-dddD(2)</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>3</td>
| |
- | <td>pSB1C3-dddD(3)</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>4</td>
| |
- | <td>1kb ladder (3µL)</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>5</td>
| |
- | <td>1kb ladder (1µL)</td>
| |
- | </tr>
| |
- | </table>
| |
- | <h4>@</h4>
| |
- | <span class="kyoto-author">Tatsui</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Lane</th>
| |
- | <th>Sample</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>1</td>
| |
- | <td>1kb ladder</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>2</td>
| |
- | <td>Blank</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>3</td>
| |
- | <td>7/18 RBA30@(pSB1C3) 2070bp</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>4</td>
| |
- | <td>Blank</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>5</td>
| |
- | <td>7/18 RBA30@(pSB1C3)</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>6</td>
| |
- | <td>Blank</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>7</td>
| |
- | <td>8/2 cloning dddD 2544bp</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>8</td>
| |
- | <td>Blank</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>9</td>
| |
- | <td>8/2 cloning dddD</td>
| |
- | </tr>
| |
- | </table>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Sample</th>
| |
- | <th>Concentration/(µg/mL)</th>
| |
- | <th>260/280</th>
| |
- | <th>260/230</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>7/18 RBS30(pSB1C3)@</td>
| |
- | <td>@</td>
| |
- | <td>1.37</td>
| |
- | <td>0.42</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>8/2 cloning dddD</td>
| |
- | <td>35</td>
| |
- | <td>1.54</td>
| |
- | <td>0.85</td>
| |
- | </tr>
| |
- | </table>
| |
- |
| |
- | <h4>Ligation</h4>
| |
- | <span class="kyoto-author">@</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th colspan=2>Vector/(µL)</th>
| |
- | <th colspan=2>Insert/(µL)</th>
| |
- | <th colspan=2>Buffer/(µL)</th>
| |
- | <th colspan=2>Ligase/(µL)</th>
| |
- | <th>MilliQ/(µL)</th>
| |
- | <th>Total/(µL)</tr>
| |
- | <tr>
| |
- | <td>@</td>
| |
- | <td>7</td>
| |
- | <td>@</td>
| |
- | <td>7</td>
| |
- | <td>2xBuffer</td>
| |
- | <td>15</td>
| |
- | <td>T4Ligase</td>
| |
- | <td>1</td>
| |
- | <td>@</td>
| |
- | <td>@</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>@</td>
| |
- | <td>7</td>
| |
- | <td>@</td>
| |
- | <td>14</td>
| |
- | <td>2xBuffer</td>
| |
- | <td>22</td>
| |
- | <td>T4Ligase</td>
| |
- | <td>1</td>
| |
- | <td>@</td>
| |
- | <td>@</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>@</td>
| |
- | <td>7</td>
| |
- | <td>@</td>
| |
- | <td>@</td>
| |
- | <td>2xBuffer</td>
| |
- | <td>15</td>
| |
- | <td>T4Ligase</td>
| |
- | <td>1</td>
| |
- | <td>7</td>
| |
- | <td>@</td>
| |
- | </tr>
| |
- | </table>
| |
- | <h4>Transformation</h4>
| |
- | <span class="kyoto-author">@</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th colspan=2>Sample/(µL)</th>
| |
- | <th colspan=2>CompetentCells/(µL)</th>
| |
- | <th>Total/(µL)</th>
| |
- | <th>Medium</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>@</td>
| |
- | <td>@</td>
| |
- | <td>@</td>
| |
- | <td>@</td>
| |
- | <td>@</td>
| |
- | <td>@</td>
| |
- | </tr>
| |
- | </table>
| |
- | </div>
| |
- |
| |
- | <div>
| |
- |
| |
- | <a name="0804" class="kyoto-jump"></a>
| |
- | <h3>8/4</h3>
| |
- |
| |
- | <h4>Colony PCR</h4>
| |
- | <span class="kyoto-author">Honda</span>
| |
- | <p>Master Mix</p>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Quick Tag/(µL)</th>
| |
- | <th>VF2/(µL)</th>
| |
- | <th>VR/(µL)</th>
| |
- | <th>MilliQ/(µL)</th>
| |
- | <th>Total/(µL)</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>40</td>
| |
- | <td>1.6</td>
| |
- | <td>1.6</td>
| |
- | <td>36.8</td>
| |
- | <td>80</td>
| |
- | </tr>
| |
- | </table>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Predenature</th>
| |
- | <th>Denature</th>
| |
- | <th>Annealing</th>
| |
- | <th>Extension</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>94@</td>
| |
- | <td>94@</td>
| |
- | <td>55@</td>
| |
- | <td>68@</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>2min</td>
| |
- | <td>30sec</td>
| |
- | <td>30sec</td>
| |
- | <td>24sec</td>
| |
- | </tr>
| |
- | </table>
| |
- | <p>Denature-Annealing-Extension 30cycle</p>
| |
- | <table>
| |
- | <tr>
| |
- | <th>1</th>
| |
- | <th>dddD+pSB1C3(1)-1</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>2</td>
| |
- | <td>dddD+pSB1C3(1)-2</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>3</td>
| |
- | <td>dddD+pSB1C3(2)-1</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>4</td>
| |
- | <td>dddD+pSB1C3(2)-2</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>5</td>
| |
- | <td>dddD+pSB1C3(2)-3</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>6</td>
| |
- | <td>dddD+pSB1C3(2)-4</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>7</td>
| |
- | <td>pSB1C3 control-1</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>8</td>
| |
- | <td>control</td>
| |
- | </tr>
| |
- | </table>
| |
- | <h4>@</h4>
| |
- | <span class="kyoto-author">Matsumoto</span>
| |
- | <table>
| |
- | <tr>
| |
- | <th>Lane</th>
| |
- | <th>Sample</th>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>1</td>
| |
- | <td>1kbp ladder</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>2</td>
| |
- | <td>dddD+pSB1C3(1)-1</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>3</td>
| |
- | <td>dddD+pSB1C3(1)-2</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>4</td>
| |
- | <td>dddD+pSB1C3(2)-1</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>5</td>
| |
- | <td>dddD+pSB1C3(2)-2</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>6</td>
| |
- | <td>dddD+pSB1C3(2)-3</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>7</td>
| |
- | <td>dddD+pSB1C3(2)-4</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>8</td>
| |
- | <td>pSB1C3 control-1</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>9</td>
| |
- | <td>control</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>10</td>
| |
- | <td>1kbp ladder</td>
| |
- | </tr>
| |
- | </table>
| |
- | </div>
| |
- |
| |
- |
| |
- |
| |
- |
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- |
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