Team:KIT-Kyoto/Notebook/Labnote
From 2014.igem.org
(100 intermediate revisions not shown) | |||
Line 4: | Line 4: | ||
{{Team:KIT-Kyoto/slidemenu}} | {{Team:KIT-Kyoto/slidemenu}} | ||
{{Team:KIT-Kyoto/navi}} | {{Team:KIT-Kyoto/navi}} | ||
+ | {{Team:KIT-Kyoto/close_navi}} | ||
{{Team:KIT-Kyoto/table}} | {{Team:KIT-Kyoto/table}} | ||
{{Team:KIT-Kyoto/Scroll}} | {{Team:KIT-Kyoto/Scroll}} | ||
Line 11: | Line 12: | ||
<!--サイドメニュー--> | <!--サイドメニュー--> | ||
<ul id="nav5"> | <ul id="nav5"> | ||
- | <li class="menuimg"><a href="/Team:KIT-Kyoto | + | <li class="menuimg"><a href="/Team:KIT-Kyoto">home</a></li> |
- | <li class="menuimg"><a href="/Team:KIT-Kyoto | + | <li class="menuimg"><a href="/Team:KIT-Kyoto/About">about us</a> |
- | <li class="menuimg"><a href="/Team:KIT-Kyoto | + | <li class="menuimg"><a href="/Team:KIT-Kyoto/Project">project</a></li> |
- | + | ||
<li class="menuimg"><a href="/Team:KIT-Kyoto/HumanPractice">policy & practices</a></li> | <li class="menuimg"><a href="/Team:KIT-Kyoto/HumanPractice">policy & practices</a></li> | ||
+ | <li class="menuimg"><a href="/Team:KIT-Kyoto/Achievement">Achievement</a></li> | ||
</ul> | </ul> | ||
<div class="active"> | <div class="active"> | ||
Line 21: | Line 22: | ||
</div> | </div> | ||
<ul class="submenu"> | <ul class="submenu"> | ||
- | <li><a href=" | + | <li class="category"> |
+ | <a href="javascript:void(0)"> | ||
+ | <font color="#143">Click Here To Enlarge</font> LabNote | ||
+ | </a> | ||
+ | </li> | ||
+ | <ul class="slidemenu"> | ||
+ | <li class="btn"> | ||
+ | <a href="">June | ||
+ | </a> | ||
+ | </li> | ||
+ | <li class="btn"> | ||
+ | <a href="">July | ||
+ | </a> | ||
+ | </li> | ||
+ | <li class="btn"> | ||
+ | <a href="">August | ||
+ | </a> | ||
+ | </li> | ||
+ | <li class="btn"> | ||
+ | <a href="">September | ||
+ | </a> | ||
+ | </li> | ||
+ | <li class="btn"> | ||
+ | <a href="">October | ||
+ | </a> | ||
+ | </li> | ||
+ | </ul> | ||
<li><a href="/Team:KIT-Kyoto/Notebook/Protocol">Protocol</a></li> | <li><a href="/Team:KIT-Kyoto/Notebook/Protocol">Protocol</a></li> | ||
</ul> | </ul> | ||
Line 29: | Line 56: | ||
<div id="container"> | <div id="container"> | ||
<div class="main-contents"> | <div class="main-contents"> | ||
+ | <div id="ln"> | ||
+ | <div class="accordion"> | ||
<h1>LabNote</h1> | <h1>LabNote</h1> | ||
- | <h2>June</h2> | + | <h2><a href="javascript:void(0);">June</a></h2> |
<div class="scroll"></div> | <div class="scroll"></div> | ||
+ | <div class="ac"> | ||
<!-- タイトル --> | <!-- タイトル --> | ||
<h3>June 10, 2014</h3> | <h3>June 10, 2014</h3> | ||
Line 43: | Line 73: | ||
<!--Miniprep--> | <!--Miniprep--> | ||
<strong>Miniprep</strong> | <strong>Miniprep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 72: | Line 103: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Miniprep終わり--> | <!--Miniprep終わり--> | ||
Line 78: | Line 110: | ||
<!--Restriction Digest--> | <!--Restriction Digest--> | ||
<strong>Restriction Digest</strong> | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 90: | Line 123: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>BL21(DE3) | + | <td>BL21(DE3)</td> |
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>BL21(DE3) | + | <td>BL21(DE3)</td> |
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>BL21(DE3) | + | <td>BL21(DE3)</td> |
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>BL21(DE3) | + | <td>BL21(DE3)</td> |
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTS62</td> |
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>BL21(DE3) | + | <td>BL21(DE3)</td> |
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td><em>Sal</em>1, <em>Not</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>BL21(DE3) | + | <td>BL21(DE3)</td> |
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td><em>Sal</em>1, <em>Not</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>BL21(DE3) | + | <td>BL21(DE3)</td> |
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td><em>Sal</em>1, <em>Not</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>BL21(DE3) | + | <td>BL21(DE3)</td> |
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTS62</td> |
- | <td> | + | <td><em>Sal</em>1, <em>Not</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Ristriction Digest--> | <!--Ristriction Digest--> | ||
<br> | <br> | ||
Line 151: | Line 185: | ||
</p> | </p> | ||
<!--本文終わり--> | <!--本文終わり--> | ||
- | |||
<!-- タイトル --> | <!-- タイトル --> | ||
<h3>June 11, 2014</h3> | <h3>June 11, 2014</h3> | ||
Line 163: | Line 196: | ||
<!--AGE--> | <!--AGE--> | ||
<strong>AGE</strong> | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
<th rowspan="2">Lane</th> | <th rowspan="2">Lane</th> | ||
- | <th colspan=" | + | <th colspan="3">Sample</th> |
<th rowspan="2">Enzyme</th> | <th rowspan="2">Enzyme</th> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
+ | <th>Marker</th> | ||
<th>Vector</th> | <th>Vector</th> | ||
<th>Insert</th> | <th>Insert</th> | ||
Line 176: | Line 211: | ||
<td>1</td> | <td>1</td> | ||
<td>λDNA-HindⅢ</td> | <td>λDNA-HindⅢ</td> | ||
- | <td | + | <td></td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>2</td> | <td>2</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>Non Treated</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>3</td> | <td>3</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>4</td> | <td>4</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td><em>Sal</em>1, <em>Not</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>5</td> | <td>5</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>Non Treated</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>6</td> | <td>6</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>7</td> | <td>7</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td><em>Sal</em>1, <em>Not</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>8</td> | <td>8</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>Non Treated</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>9</td> | <td>9</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>10</td> | <td>10</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTS3</td> |
- | <td> | + | <td><em>Sal</em>1, <em>Not</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>11</td> | <td>11</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>Non Treated</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>12</td> | <td>12</td> | ||
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
+ | </tr> | ||
+ | <tr> | ||
+ | <td>13</td> | ||
+ | <td></td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTS62</td> | ||
+ | <td><em>Sal</em>1, <em>Not</em>1</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--AGE--> | <!--AGE--> | ||
+ | <img src="/wiki/images/8/88/Kit_June11_c.jpg"> | ||
<br> | <br> | ||
<br> | <br> | ||
Line 262: | Line 319: | ||
<!--Transformation--> | <!--Transformation--> | ||
<strong>Transformation</strong> | <strong>Transformation</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 291: | Line 349: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Transformation終わり--> | <!--Transformation終わり--> | ||
<br> | <br> | ||
Line 305: | Line 364: | ||
<!--Colony Isolation--> | <!--Colony Isolation--> | ||
<strong>Colony Isolation</strong><br> | <strong>Colony Isolation</strong><br> | ||
- | Select 6 colonies per plate from 4 plates from June | + | Select 6 colonies per plate from 4 plates from June 12.<br> |
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 337: | Line 397: | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTS62</td> |
<td>6-a, 6-b</td> | <td>6-a, 6-b</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Colony Isolation--> | <!--Colony Isolation--> | ||
<br> | <br> | ||
Line 360: | Line 421: | ||
<!--Miniprep--> | <!--Miniprep--> | ||
<strong>Miniprep</strong> | <strong>Miniprep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 409: | Line 471: | ||
</table> | </table> | ||
+ | </div> | ||
<!--Miniprep終わり--> | <!--Miniprep終わり--> | ||
<br> | <br> | ||
<!--Miniprep--> | <!--Miniprep--> | ||
<strong>Miniprep</strong> | <strong>Miniprep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 443: | Line 507: | ||
</table> | </table> | ||
+ | </div> | ||
<!--Miniprep終わり--> | <!--Miniprep終わり--> | ||
<br> | <br> | ||
<!--Restriction Digest--> | <!--Restriction Digest--> | ||
<strong>Restriction Digest</strong> | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 462: | Line 528: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 469: | Line 535: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 476: | Line 542: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 483: | Line 549: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 490: | Line 556: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 497: | Line 563: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 504: | Line 570: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 511: | Line 577: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 518: | Line 584: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 525: | Line 591: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 532: | Line 598: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 539: | Line 605: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Ristriction Digest--> | <!--Ristriction Digest--> | ||
Line 548: | Line 615: | ||
<!--AGE--> | <!--AGE--> | ||
<strong>AGE</strong> | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
<th rowspan="2">Lane</th> | <th rowspan="2">Lane</th> | ||
- | <th colspan=" | + | <th colspan="3">Sample</th> |
<th rowspan="2">Insert</th> | <th rowspan="2">Insert</th> | ||
<th rowspan="2">Enzyme</th> | <th rowspan="2">Enzyme</th> | ||
Line 557: | Line 625: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
+ | <th>Marker</th> | ||
<th>Host</th> | <th>Host</th> | ||
<th>Vector</th> | <th>Vector</th> | ||
Line 563: | Line 632: | ||
<td>1</td> | <td>1</td> | ||
<td>1kb Ladder</td> | <td>1kb Ladder</td> | ||
- | <td | + | <td></td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
<tr> | <tr> | ||
<td>2</td> | <td>2</td> | ||
<td>λDNA-HindⅢ</td> | <td>λDNA-HindⅢ</td> | ||
- | <td | + | <td></td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
<tr> | <tr> | ||
<td>3</td> | <td>3</td> | ||
+ | <td></td> | ||
<td>1-a</td> | <td>1-a</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>4</td> | <td>4</td> | ||
+ | <td></td> | ||
<td>1-b</td> | <td>1-b</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>5</td> | <td>5</td> | ||
+ | <td></td> | ||
<td>2-a</td> | <td>2-a</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>6</td> | <td>6</td> | ||
+ | <td></td> | ||
<td>2-b</td> | <td>2-b</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>7</td> | <td>7</td> | ||
+ | <td></td> | ||
<td>3-a</td> | <td>3-a</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>8</td> | <td>8</td> | ||
+ | <td></td> | ||
<td>3-b</td> | <td>3-b</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<td>9</td> | <td>9</td> | ||
+ | <td></td> | ||
<td>6-a</td> | <td>6-a</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
<tr> | <tr> | ||
<td>10</td> | <td>10</td> | ||
+ | <td></td> | ||
<td>6-b</td> | <td>6-b</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>11</td> | <td>11</td> | ||
+ | <td></td> | ||
<td>BL21(DE3)</td> | <td>BL21(DE3)</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>12</td> | <td>12</td> | ||
+ | <td></td> | ||
<td>BL21(DE3)</td> | <td>BL21(DE3)</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>13</td> | <td>13</td> | ||
+ | <td></td> | ||
<td>BL21(DE3)</td> | <td>BL21(DE3)</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>14</td> | <td>14</td> | ||
+ | <td></td> | ||
<td>BL21(DE3)</td> | <td>BL21(DE3)</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--AGE--> | <!--AGE--> | ||
<br> | <br> | ||
- | + | Apply λDNA-HindⅢ to compare and contrast AGE of July 14 and June 11. | |
<br> | <br> | ||
+ | <img src="/wiki/images/b/b6/Kit_June14_c.jpg"> | ||
<br> | <br> | ||
</p> | </p> | ||
Line 682: | Line 773: | ||
<!--Preculture--> | <!--Preculture--> | ||
<strong>Preculture</strong> | <strong>Preculture</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 712: | Line 804: | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
- | <td | + | <td>pGEX6P-2</td> |
+ | <td></td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Preculture終わり--> | <!--Preculture終わり--> | ||
<br> | <br> | ||
<!--Main Culture--> | <!--Main Culture--> | ||
<strong>Main Culture</strong> | <strong>Main Culture</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 731: | Line 826: | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
- | <td | + | <td>pGEX6P-2</td> |
+ | <td>CuMTSE1</td> | ||
<td>Induced</td> | <td>Induced</td> | ||
</tr> | </tr> | ||
+ | <tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTSE2</td> | ||
+ | <td>Induced</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>Induced</td> | ||
+ | <tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTS62</td> | ||
+ | <td>Induced</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td></td> | ||
+ | <td>Induced</td> | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
+ | <td>Non-induced</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTSE2</td> | ||
+ | <td>Non-induced</td> | ||
+ | </tr><tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>Non-induced</td> | ||
+ | </tr><tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTS62</td> | ||
+ | <td>Non-induced</td> | ||
+ | </tr><tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td></td> | ||
+ | <td>Non-induced</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <!--Main Culture終わり--> | ||
+ | <br> | ||
+ | </p> | ||
+ | <!--本文終わり--> | ||
+ | <br> | ||
+ | <!-- タイトル --> | ||
+ | <h3>June 18, 2014</h3> | ||
+ | <!-- タイトル終わり --> | ||
+ | |||
+ | <!-- 本文開始 --> | ||
+ | <p class="sentence"> | ||
+ | <!-- 実験者名 --> | ||
+ | YAMAJI<br> | ||
+ | <!-- 実験者名終わり --> | ||
+ | <!--Preculture--> | ||
+ | <strong>Preculture</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="2">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | </tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTSE2</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <!--Preculture終わり--> | ||
+ | <br> | ||
+ | <!--Main Culture--> | ||
+ | <strong>Main Culture</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="2">Sample</th> | ||
+ | <th rowspan="2">IPTG</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td></td> | ||
<td>Induced</td> | <td>Induced</td> | ||
</tr> | </tr> | ||
Line 743: | Line 942: | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE1</td> |
<td>Induced</td> | <td>Induced</td> | ||
+ | </tr> | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
- | <td | + | <td>pGEX6P-2</td> |
- | <td>Non- | + | <td>CuMTSE2</td> |
+ | <td>Induced</td> | ||
+ | <tr> | ||
+ | <td>BL21(DE3)pLysS</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td></td> | ||
+ | <td>Non-induced</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE1</td> |
- | <td>Non- | + | <td>Non-induced</td> |
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td>Non- | + | <td>Non-induced</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Main Culture終わり--> | <!--Main Culture終わり--> | ||
<br> | <br> | ||
- | <!--Protein Extraction (<em>E.coli</em>)--> | + | <!--Protein Extraction (<em>E. coli</em>)--> |
- | <strong>Protein Extraction (<em>E.coli</em>)</strong> | + | <strong>Protein Extraction (<em>E. coli</em>)</strong> |
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 778: | Line 986: | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
- | <td | + | <td>pGEX6P-2</td> |
+ | <td></td> | ||
<td>Induced</td> | <td>Induced</td> | ||
</tr> | </tr> | ||
Line 790: | Line 999: | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE2</td> |
<td>Induced</td> | <td>Induced</td> | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
- | <td | + | <td>pGEX6P-2</td> |
- | <td>Non- | + | <td></td> |
+ | <td>Non-induced</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE1</td> |
- | <td>Non- | + | <td>Non-induced</td> |
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td>Non- | + | <td>Non-induced</td> |
</tr> | </tr> | ||
</table> | </table> | ||
- | <!--Protein Extraction (<em>E.coli</em>)終わり--> | + | </div> |
+ | <!--Protein Extraction (<em>E. coli</em>)終わり--> | ||
<br> | <br> | ||
<!--Preparations for SDS-PAGE--> | <!--Preparations for SDS-PAGE--> | ||
Line 817: | Line 1,028: | ||
<!--Preparations for SDS-PAGE終わり--> | <!--Preparations for SDS-PAGE終わり--> | ||
<br> | <br> | ||
+ | <img src="/wiki/images/0/09/Kit_June18_c.jpg"> | ||
<br> | <br> | ||
<br> | <br> | ||
Line 833: | Line 1,045: | ||
<!--SDS-PAGE--> | <!--SDS-PAGE--> | ||
<strong>SDS-PAGE</strong> | <strong>SDS-PAGE</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 845: | Line 1,058: | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
- | <td | + | <td>pGEX6P-2</td> |
+ | <td></td> | ||
<td>Induced</td> | <td>Induced</td> | ||
</tr> | </tr> | ||
Line 857: | Line 1,071: | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE2</td> |
<td>Induced</td> | <td>Induced</td> | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
- | <td | + | <td>pGEX6P-2</td> |
- | <td>Non- | + | <td></td> |
+ | <td>Non-induced</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE1</td> |
- | <td>Non- | + | <td>Non-induced</td> |
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td>Non- | + | <td>Non-induced</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--SDS-PAGE終わり--> | <!--SDS-PAGE終わり--> | ||
Line 894: | Line 1,110: | ||
<!--Miniprep--> | <!--Miniprep--> | ||
<strong>Miniprep</strong> | <strong>Miniprep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 906: | Line 1,123: | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 914: | Line 1,131: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Mineprep終わり--> | <!--Mineprep終わり--> | ||
<br> | <br> | ||
<!--PCR--> | <!--PCR--> | ||
<strong>PCR</strong> | <strong>PCR</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 931: | Line 1,150: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE1</td> |
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE2</td> |
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
<tr> | <tr> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--PCR終わり--> | <!--PCR終わり--> | ||
<br> | <br> | ||
<!--AGE--> | <!--AGE--> | ||
<strong>AGE</strong> | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
<th rowspan="2">Lane</th> | <th rowspan="2">Lane</th> | ||
- | <th colspan=" | + | <th colspan="3">Sample</th> |
<th rowspan="2">Insert</th> | <th rowspan="2">Insert</th> | ||
<th rowspan="2">Primer</th> | <th rowspan="2">Primer</th> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
+ | <th>Marker</th> | ||
<th>Host</th> | <th>Host</th> | ||
<th>Vector</th> | <th>Vector</th> | ||
Line 970: | Line 1,192: | ||
<td>1</td> | <td>1</td> | ||
<td>1kb Ladder</td> | <td>1kb Ladder</td> | ||
- | <td | + | <td></td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>2</td> | <td>2</td> | ||
+ | <td></td> | ||
<td>BY4247</td> | <td>BY4247</td> | ||
- | <td | + | <td>p427-TEF</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>3</td> | <td>3</td> | ||
+ | <td></td> | ||
<td></td> | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<tr> | <tr> | ||
<td>4</td> | <td>4</td> | ||
+ | <td></td> | ||
<td></td> | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE1</td> |
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>5</td> | <td>5</td> | ||
+ | <td></td> | ||
<td></td> | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE2</td> |
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>6</td> | <td>6</td> | ||
+ | <td></td> | ||
<td></td> | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--AGE終わり--> | <!--AGE終わり--> | ||
<br> | <br> | ||
+ | <img src="/wiki/images/2/29/Kit_June21_c.jpg"> | ||
<br> | <br> | ||
</p> | </p> | ||
Line 1,023: | Line 1,257: | ||
<!--Preculture--> | <!--Preculture--> | ||
<strong>Preculture</strong> | <strong>Preculture</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,033: | Line 1,268: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>BL21(DE3)pLysS</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,045: | Line 1,282: | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Preculture終わり--> | <!--Preculture終わり--> | ||
<br> | <br> | ||
Line 1,061: | Line 1,299: | ||
<!-- 実験者名終わり --> | <!-- 実験者名終わり --> | ||
<br> | <br> | ||
- | + | Cultivate following samples in Big Scale at 30℃ overnight in shaken culture. | |
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,073: | Line 1,312: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>BL21(DE3)pLysS</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,086: | Line 1,327: | ||
<tr> | <tr> | ||
</table> | </table> | ||
+ | </div> | ||
<br> | <br> | ||
- | <!--DNA | + | <!--DNA Refinement 1--> |
- | <strong>DNA | + | <strong>DNA Refinement 1</strong> |
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,101: | Line 1,344: | ||
<tr> | <tr> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>CuMTSE1</td> |
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</table> | </table> | ||
- | <!--DNA | + | </div> |
+ | <!--DNA Refinement 1終わり--> | ||
<br> | <br> | ||
Line 1,113: | Line 1,357: | ||
<!--Restriction Digest--> | <!--Restriction Digest--> | ||
<strong>Restriction Digest</strong> | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,127: | Line 1,372: | ||
<td></td> | <td></td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td>CuMTSE1</td> |
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td></td> |
+ | <td><em>Spe</em>1, <em>Xho</em>1</td> | ||
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Ristriction Digest--> | <!--Ristriction Digest--> | ||
<br> | <br> | ||
- | <!--DNA | + | <!--DNA Refinement 2--> |
- | <strong>DNA | + | <strong>DNA Refinement 2</strong> |
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,164: | Line 1,412: | ||
<td></td> | <td></td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td>CuMTSE1</td> |
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td></td> |
+ | <td><em>Spe</em>1, <em>Xho</em>1</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
- | <!--DNA | + | </div> |
+ | <!--DNA Refinement 2--> | ||
<br> | <br> | ||
Line 1,185: | Line 1,435: | ||
<!--Main Culture--> | <!--Main Culture--> | ||
<strong>Main Culture</strong> | <strong>Main Culture</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,217: | Line 1,468: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td></td> | <td></td> | ||
- | <td>Non- | + | <td>Non-induced</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,223: | Line 1,474: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td>Non- | + | <td>Non-induced</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,229: | Line 1,480: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td>Non- | + | <td>Non-induced</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Main Culture--> | <!--Main Culture--> | ||
- | + | <img src="/wiki/images/1/1f/Kit_June25_c.jpg"> | |
<br> | <br> | ||
<!--Ligation--> | <!--Ligation--> | ||
<strong>Ligation</strong> | <strong>Ligation</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,250: | Line 1,503: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
DNA ligase: DNA Ligation Kit ‹Mighty Mix› (TAKARA BIO INC.) | DNA ligase: DNA Ligation Kit ‹Mighty Mix› (TAKARA BIO INC.) | ||
<!--Ligation--> | <!--Ligation--> | ||
Line 1,275: | Line 1,529: | ||
<!--Transformation--> | <!--Transformation--> | ||
<strong>Transformation</strong> | <strong>Transformation</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,289: | Line 1,544: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,295: | Line 1,550: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,301: | Line 1,556: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,307: | Line 1,562: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Transformation--> | <!--Transformation--> | ||
<br> | <br> | ||
Line 1,330: | Line 1,586: | ||
Isolate each colony from the plates which were inoculated on June 25. | Isolate each colony from the plates which were inoculated on June 25. | ||
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,345: | Line 1,602: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>1~4</td> | <td>1~4</td> | ||
</tr> | </tr> | ||
Line 1,352: | Line 1,609: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>a~h</td> | <td>a~h</td> | ||
</tr> | </tr> | ||
Line 1,359: | Line 1,616: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>ア~タ</td> | <td>ア~タ</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Colony Isolation--> | <!--Colony Isolation--> | ||
<br> | <br> | ||
Line 1,378: | Line 1,636: | ||
<!--Colony Sweep--> | <!--Colony Sweep--> | ||
<strong>Colony Sweep</strong> | <strong>Colony Sweep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,396: | Line 1,655: | ||
<tr> | <tr> | ||
<td>2</td> | <td>2</td> | ||
- | <td | + | <td>1</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>3</td> | <td>3</td> | ||
- | <td | + | <td>2</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>4</td> | <td>4</td> | ||
- | <td | + | <td>3</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>5</td> | <td>5</td> | ||
- | <td | + | <td>4</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>6</td> | <td>6</td> | ||
- | <td | + | <td>a</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>7</td> | <td>7</td> | ||
- | <td | + | <td>b</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>8</td> | <td>8</td> | ||
- | <td | + | <td>c</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>9</td> | <td>9</td> | ||
- | <td | + | <td>d</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>10</td> | <td>10</td> | ||
- | <td | + | <td>e</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>11</td> | <td>11</td> | ||
- | <td | + | <td>f</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>12</td> | <td>12</td> | ||
- | <td | + | <td>g</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>13</td> | <td>13</td> | ||
- | <td | + | <td>h</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Colony Sweep--> | <!--Colony Sweep--> | ||
+ | <img src="/wiki/images/4/4d/Kit_June27-1_c.jpg"> | ||
<br> | <br> | ||
<!--Colony Sweep--> | <!--Colony Sweep--> | ||
<strong>Colony Sweep</strong> | <strong>Colony Sweep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,466: | Line 1,752: | ||
<tr> | <tr> | ||
<td>2</td> | <td>2</td> | ||
- | <td | + | <td>ア</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>3</td> | <td>3</td> | ||
- | <td | + | <td>イ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>4</td> | <td>4</td> | ||
- | <td | + | <td>ウ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>5</td> | <td>5</td> | ||
- | <td | + | <td>エ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>6</td> | <td>6</td> | ||
- | <td | + | <td>オ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>7</td> | <td>7</td> | ||
- | <td | + | <td>カ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>8</td> | <td>8</td> | ||
- | <td | + | <td>キ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>9</td> | <td>9</td> | ||
- | <td | + | <td>ク</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>10</td> | <td>10</td> | ||
- | <td | + | <td>ケ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>11</td> | <td>11</td> | ||
- | <td | + | <td>コ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>12</td> | <td>12</td> | ||
- | <td | + | <td>サ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>13</td> | <td>13</td> | ||
- | <td | + | <td>シ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>14</td> | <td>14</td> | ||
- | <td | + | <td>ス</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>15</td> | <td>15</td> | ||
- | <td | + | <td>セ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>16</td> | <td>16</td> | ||
- | <td | + | <td>ソ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>17</td> | <td>17</td> | ||
- | <td | + | <td>タ</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Colony Sweep--> | <!--Colony Sweep--> | ||
+ | <img src="/wiki/images/4/46/Kit_June27-2_c.jpg"> | ||
<br> | <br> | ||
Identified targeted genes in 4, c, h, イ, カ, セ | Identified targeted genes in 4, c, h, イ, カ, セ | ||
Line 1,538: | Line 1,858: | ||
<!--Miniprep--> | <!--Miniprep--> | ||
<strong>Miniprep</strong> | <strong>Miniprep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,563: | Line 1,884: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Miniprep終わり--> | <!--Miniprep終わり--> | ||
- | |||
<br> | <br> | ||
Line 1,570: | Line 1,891: | ||
<!--Restriction Digest--> | <!--Restriction Digest--> | ||
<strong>Restriction Digest</strong> | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,581: | Line 1,903: | ||
<tr> | <tr> | ||
<td>4</td> | <td>4</td> | ||
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>C</td> | <td>C</td> | ||
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>H</td> | <td>H</td> | ||
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
<tr> | <tr> | ||
<td>イ</td> | <td>イ</td> | ||
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>カ</td> | <td>カ</td> | ||
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Restriction Digest終わり--> | <!--Restriction Digest終わり--> | ||
Line 1,610: | Line 1,933: | ||
<!--AGE--> | <!--AGE--> | ||
<strong>AGE</strong> | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
<th rowspan="2">Lane</th> | <th rowspan="2">Lane</th> | ||
- | <th colspan=" | + | <th colspan="3">Sample</th> |
<th rowspan="2">Primer</th> | <th rowspan="2">Primer</th> | ||
<th rowspan="2">Enzyme</th> | <th rowspan="2">Enzyme</th> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
+ | <th>Marker</th> | ||
<th>Host</th> | <th>Host</th> | ||
<th>Insert</th> | <th>Insert</th> | ||
Line 1,623: | Line 1,948: | ||
<tr> | <tr> | ||
<td>1</td> | <td>1</td> | ||
- | <td> | + | <td>1kb Ladder</td> |
- | <td | + | <td></td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>2</td> |
+ | <td></td> | ||
+ | <td></td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>3</td> | <td>3</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td>4</td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1, <em>Xho</em>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>4</td> | <td>4</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td>4</td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1, <em>Xho</em>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>5</td> | <td>5</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td>4</td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>Non Treated</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>6</td> |
+ | <td></td> | ||
+ | <td></td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>7</td> | <td>7</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td>c</td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1, <em>Xho</em>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>8</td> | <td>8</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td>c</td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1, <em>Xho</em>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>9</td> | <td>9</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td>c</td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>Non Treated</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>10</td> | <td>10</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td>h</td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1, <em>Xho</em>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>11</td> | <td>11</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td>h</td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1, <em>Xho</em>1</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>12</td> | <td>12</td> | ||
- | <td | + | <td></td> |
- | <td> | + | <td>h</td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>Non Treated</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--AGE--> | <!--AGE--> | ||
+ | <img src="/wiki/images/9/95/June27-3_c.jpg"> | ||
<br> | <br> | ||
<!--Transformation (electroportation)--> | <!--Transformation (electroportation)--> | ||
<strong>Transformation (electroportation)</strong> | <strong>Transformation (electroportation)</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,702: | Line 2,065: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,708: | Line 2,071: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Transformation (electroportation)--> | <!--Transformation (electroportation)--> | ||
Line 1,732: | Line 2,096: | ||
Divide each of 2 plates into 8 sections and inoculate following samples on them. | Divide each of 2 plates into 8 sections and inoculate following samples on them. | ||
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,747: | Line 2,112: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>A~H</td> | <td>A~H</td> | ||
</tr> | </tr> | ||
Line 1,754: | Line 2,119: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>一~ハ</td> | <td>一~ハ</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
Cultivate 17 samples in a small scale at 30℃ for 6 hours in shaken culture. | Cultivate 17 samples in a small scale at 30℃ for 6 hours in shaken culture. | ||
<!--Inoculation--> | <!--Inoculation--> | ||
Line 1,767: | Line 2,133: | ||
</p> | </p> | ||
<!--本文終わり--> | <!--本文終わり--> | ||
+ | </div> | ||
+ | </div> | ||
- | <h2>July</h2> | + | <div class="accordion"> |
+ | <h2><a href="javascript:void(0);">July</a></h2> | ||
<div class="scroll"></div> | <div class="scroll"></div> | ||
+ | <div class="ac"> | ||
Line 1,781: | Line 2,151: | ||
KOBAYASHI<br> | KOBAYASHI<br> | ||
<!-- 実験者名終わり --> | <!-- 実験者名終わり --> | ||
- | <!--Protein Extraction (<em> | + | <!--Protein Extraction (<em>S. cerevisiae</em>)--> |
- | <strong>Protein Extraction (<em> | + | <strong>Protein Extraction (<em>S. cerevisiae</em>)</strong> |
- | < | + | <div class="m_table"> |
- | + | ||
- | + | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,800: | Line 2,168: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,806: | Line 2,174: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
- | <!--Protein Extraction (<em> | + | </div> |
+ | <!--Protein Extraction (<em>S. cerevisiae</em>)--> | ||
<br> | <br> | ||
Line 1,826: | Line 2,195: | ||
<strong>Miniprep</strong> | <strong>Miniprep</strong> | ||
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,838: | Line 2,208: | ||
<tr> | <tr> | ||
<td>BY4247</td> | <td>BY4247</td> | ||
- | <td | + | <td>p427-TEF</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,844: | Line 2,216: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,850: | Line 2,222: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Miniprep--> | <!--Miniprep--> | ||
Line 1,859: | Line 2,232: | ||
<!--Restriction Digest--> | <!--Restriction Digest--> | ||
<strong>Restriction Digest</strong> | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,874: | Line 2,248: | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 1,880: | Line 2,254: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 1,887: | Line 2,261: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Eco</em>R1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 1,895: | Line 2,269: | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td><em>Sac</em>1, <em>Kpn</em>1</td> |
<td>Buffer L</td> | <td>Buffer L</td> | ||
</tr> | </tr> | ||
Line 1,901: | Line 2,275: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Sac</em>1, <em>Kpn</em>1</td> |
<td>Buffer L</td> | <td>Buffer L</td> | ||
</tr> | </tr> | ||
Line 1,908: | Line 2,282: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Sac</em>1, <em>Kpn</em>1</td> |
+ | <td>Buffer L</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <!--Restriction Digest--> | ||
+ | |||
+ | <br> | ||
+ | |||
+ | <!--AGE--> | ||
+ | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="4">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Marker</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>Non Treated</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F1, pGEX6P-2R</td> | ||
+ | <td>Non Treated</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | <td>Non Treated</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>5</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>6</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F1, pGEX6P-2R</td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>7</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>8</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>9</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Kpn</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>10</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F1, pGEX6P-2R</td> | ||
+ | <td><em>Kpn</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>11</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | <td><em>Kpn</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>12</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>13</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Sac</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>14</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F1, pGEX6P-2R</td> | ||
+ | <td><em>Sac</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>15</td> | ||
+ | <td></td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | <td><em>Sac</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <!--AGE--> | ||
+ | <img src="/wiki/images/8/82/Kit_July2-1_c.jpg"> | ||
+ | <br> | ||
+ | |||
+ | <!--Restriction Digest--> | ||
+ | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | <th rowspan="2">Buffer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F1, pGEX6P-2R</td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td></td> | ||
+ | <td><em>Sac</em>1</td> | ||
+ | <td>Buffer L</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F1, pGEX6P-2R</td> | ||
+ | <td><em>Sac</em>1</td> | ||
+ | <td>Buffer L</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE1</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | <td><em>Sac</em>1</td> | ||
<td>Buffer L</td> | <td>Buffer L</td> | ||
</tr> | </tr> | ||
Line 1,916: | Line 2,494: | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td><em>Kpn</em>1</td> |
<td>Buffer L</td> | <td>Buffer L</td> | ||
</tr> | </tr> | ||
Line 1,922: | Line 2,500: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Kpn</em>1</td> |
<td>Buffer L</td> | <td>Buffer L</td> | ||
</tr> | </tr> | ||
Line 1,929: | Line 2,507: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Kpn</em>1</td> |
<td>Buffer L</td> | <td>Buffer L</td> | ||
</tr> | </tr> | ||
Line 1,937: | Line 2,515: | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 1,943: | Line 2,521: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
Line 1,950: | Line 2,528: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Buffer H</td> | <td>Buffer H</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Restriction Digest--> | <!--Restriction Digest--> | ||
Line 1,961: | Line 2,540: | ||
<!--AGE--> | <!--AGE--> | ||
<strong>AGE</strong> | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 1,968: | Line 2,548: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <th></th> | + | <th>Marker</th> |
<th>Vector</th> | <th>Vector</th> | ||
<th>Insert</th> | <th>Insert</th> | ||
Line 1,976: | Line 2,556: | ||
<td>1</td> | <td>1</td> | ||
<td>1kb Ladder</td> | <td>1kb Ladder</td> | ||
- | <td | + | <td></td> |
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 1,984: | Line 2,567: | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td><em>Eco</em>R1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>3</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Eco</em>R1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>4</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Eco</em>R1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>5</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td><em>Kpn</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>6</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Kpn</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>7</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Kpn</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>8</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td><em>Sac</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>9</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Sac</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>10</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Sac</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>11</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>12</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>13</td> |
+ | <td></td> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--AGE--> | <!--AGE--> | ||
- | + | <img src="/wiki/images/c/c1/Kit_July2-2_c.jpg"> | |
<br> | <br> | ||
<br> | <br> | ||
Line 2,081: | Line 2,676: | ||
<strong>PCR</strong> | <strong>PCR</strong> | ||
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,094: | Line 2,690: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
Line 2,100: | Line 2,696: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
Line 2,106: | Line 2,702: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
Line 2,112: | Line 2,708: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
Line 2,118: | Line 2,714: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
Line 2,124: | Line 2,720: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--PCR--> | <!--PCR--> | ||
<br> | <br> | ||
- | <!--Protein Extraction (<em> | + | <!--Protein Extraction (<em>S. cerevisiae</em>)--> |
- | <strong>Protein Extraction (<em> | + | <strong>Protein Extraction (<em>S. cerevisiae</em>)</strong> |
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,152: | Line 2,750: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>A</td> |
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>六</td> |
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
- | <!--Protein Extraction (<em> | + | </div> |
+ | <!--Protein Extraction (<em>S. cerevisiae</em>)--> | ||
<br> | <br> | ||
Line 2,180: | Line 2,779: | ||
<!--AGE--> | <!--AGE--> | ||
<strong>AGE</strong> | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
<th rowspan="2">Lane</th> | <th rowspan="2">Lane</th> | ||
- | <th colspan="4">Sample</th> | + | <th colspan="4">Sample/Marker</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <th> | + | <th>Marker</th> |
<th>Vector</th> | <th>Vector</th> | ||
<th>Insert</th> | <th>Insert</th> | ||
Line 2,194: | Line 2,794: | ||
<td>1</td> | <td>1</td> | ||
<td>1kb Ladder</td> | <td>1kb Ladder</td> | ||
- | <td | + | <td></td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 2,201: | Line 2,803: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>3</td> |
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
- | <td> | + | <td>CuMTSE2</td> |
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>4</td> |
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>5</td> |
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>6</td> |
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>7</td> |
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--AGE--> | <!--AGE--> | ||
- | + | <img src="/wiki/images/9/94/Kit_July4-1_c.jpg"> | |
<br> | <br> | ||
Line 2,241: | Line 2,849: | ||
<strong>PCR</strong> | <strong>PCR</strong> | ||
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,254: | Line 2,863: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
Line 2,260: | Line 2,869: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
Line 2,266: | Line 2,875: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
Line 2,272: | Line 2,881: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
Line 2,278: | Line 2,887: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
Line 2,284: | Line 2,893: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<td>KOD-FX-NEO</td> | <td>KOD-FX-NEO</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--PCR--> | <!--PCR--> | ||
Line 2,294: | Line 2,904: | ||
<!--AGE--> | <!--AGE--> | ||
<strong>AGE</strong> | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,300: | Line 2,911: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <th></th> | + | <th>Marker</th> |
<th>Vector</th> | <th>Vector</th> | ||
<th>Insert</th> | <th>Insert</th> | ||
Line 2,308: | Line 2,919: | ||
<td>1</td> | <td>1</td> | ||
<td>1kb Ladder</td> | <td>1kb Ladder</td> | ||
- | <td | + | <td></td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 2,315: | Line 2,928: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>3</td> |
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>1kb Ladder+K83:N91</td> | <td>1kb Ladder+K83:N91</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>4</td> |
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>5</td> |
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>6</td> |
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td | + | <td>7</td> |
+ | <td></td> | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--AGE--> | <!--AGE--> | ||
- | + | <img src="/wiki/images/d/d4/Kit_July4-2_c.jpg"> | |
<br> | <br> | ||
<br> | <br> | ||
Line 2,365: | Line 2,984: | ||
<strong>SDS-PAGE</strong> | <strong>SDS-PAGE</strong> | ||
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,377: | Line 2,997: | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
- | <td | + | <td>pGEX6P-2</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 2,383: | Line 3,005: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 2,389: | Line 3,011: | ||
<td>pGEX6P-2</td> | <td>pGEX6P-2</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<tr> | <tr> | ||
<td>BY4247</td> | <td>BY4247</td> | ||
- | <td | + | <td>p427-TEF</td> |
+ | <td></td> | ||
+ | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>A</td> |
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE1</td> | <td>CuMTSE1</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>六</td> |
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
- | <td> | + | <td>CuMTSE1</td> |
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--SDS-PAGE--> | <!--SDS-PAGE--> | ||
Line 2,422: | Line 3,047: | ||
SOGA<br> | SOGA<br> | ||
<!-- 実験者名終わり --> | <!-- 実験者名終わり --> | ||
- | <!--DNA | + | <!--DNA Refinement 1--> |
- | <strong>DNA | + | <strong>DNA Refinement 1</strong> |
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,434: | Line 3,060: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>CuMTSE2</td> |
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
<tr> | <tr> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
</tr> | </tr> | ||
</table> | </table> | ||
- | <!--DNA | + | </div> |
+ | <!--DNA Refinement 1--> | ||
<br> | <br> | ||
Line 2,455: | Line 3,082: | ||
<!--Restriction Digest--> | <!--Restriction Digest--> | ||
<strong>Restriction Digest</strong> | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,469: | Line 3,097: | ||
<td></td> | <td></td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
Line 2,476: | Line 3,104: | ||
<td></td> | <td></td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
Line 2,483: | Line 3,111: | ||
<td></td> | <td></td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
<td></td> | <td></td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
Line 2,496: | Line 3,124: | ||
<td></td> | <td></td> | ||
<td></td> | <td></td> | ||
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Ristriction Digest--> | <!--Ristriction Digest--> | ||
<br> | <br> | ||
- | <!--DNA | + | <!--DNA Refinement 2--> |
- | <strong>DNA | + | <strong>DNA Refinement 2</strong> |
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,517: | Line 3,147: | ||
<tr> | <tr> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
</table> | </table> | ||
- | <!--DNA | + | </div> |
+ | <!--DNA Refinement 2--> | ||
<br> | <br> | ||
Line 2,532: | Line 3,163: | ||
<!--Ligation--> | <!--Ligation--> | ||
<strong>Ligation</strong> | <strong>Ligation</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,545: | Line 3,177: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<br> | <br> | ||
DNA ligase: DNA Ligation Kit ‹Mighty Mix› (TAKARA BIO INC.) | DNA ligase: DNA Ligation Kit ‹Mighty Mix› (TAKARA BIO INC.) | ||
<br> | <br> | ||
- | <!--DNA | + | <!--DNA Refinement 2--> |
<br> | <br> | ||
- | <!--Transformation (<em>E.coli</em>)--> | + | <!--Transformation (<em>E. coli</em>)--> |
- | <strong>Transformation (<em>E.coli</em>)</strong> | + | <strong>Transformation (<em>E. coli</em>)</strong> |
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,580: | Line 3,214: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 2,587: | Line 3,221: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 2,594: | Line 3,228: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 2,601: | Line 3,235: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
</table> | </table> | ||
- | <!--Transformation (<em>E.coli</em>)--> | + | </div> |
+ | <!--Transformation (<em>E. coli</em>)--> | ||
</p> | </p> | ||
<!--本文終わり--> | <!--本文終わり--> | ||
Line 2,626: | Line 3,261: | ||
Isolate each colony from the plates which were inoculated on July 6. | Isolate each colony from the plates which were inoculated on July 6. | ||
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,641: | Line 3,277: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>1~16</td> | <td>1~16</td> | ||
</tr> | </tr> | ||
Line 2,648: | Line 3,284: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>ア~エ</td> | <td>ア~エ</td> | ||
</tr> | </tr> | ||
Line 2,655: | Line 3,291: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
<td>a~p</td> | <td>a~p</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Colony Isolation--> | <!--Colony Isolation--> | ||
<br> | <br> | ||
- | <!--Transformation (<em>E.coli</em>)--> | + | <!--Transformation (<em>E. coli</em>)--> |
- | <strong>Transformation (<em>E.coli</em>)</strong> | + | <strong>Transformation (<em>E. coli</em>)</strong> |
<br> | <br> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,681: | Line 3,319: | ||
<td>p427-TEF</td> | <td>p427-TEF</td> | ||
<td>CuMTSE2</td> | <td>CuMTSE2</td> | ||
- | <td> | + | <td>pGEX6P-2F2, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
</table> | </table> | ||
- | <!--Transformation (<em>E.coli</em>)--> | + | </div> |
+ | <!--Transformation (<em>E. coli</em>)--> | ||
<br> | <br> | ||
Line 2,701: | Line 3,340: | ||
<!--Colony Sweep--> | <!--Colony Sweep--> | ||
<strong>Colony Sweep</strong> | <strong>Colony Sweep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,796: | Line 3,436: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Colony Sweep--> | <!--Colony Sweep--> | ||
- | + | <img src="/wiki/images/e/e6/Kit_July8-1_c.jpg"> | |
<br> | <br> | ||
<!--Colony Sweep--> | <!--Colony Sweep--> | ||
<strong>Colony Sweep</strong> | <strong>Colony Sweep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,837: | Line 3,479: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Colony Sweep--> | <!--Colony Sweep--> | ||
- | + | <img src="/wiki/images/c/c1/Kit_July8-2_c.jpg"> | |
<br> | <br> | ||
<!--Colony Sweep--> | <!--Colony Sweep--> | ||
<strong>Colony Sweep</strong> | <strong>Colony Sweep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,938: | Line 3,582: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Colony Sweep--> | <!--Colony Sweep--> | ||
- | + | <img src="/wiki/images/c/c3/Kit_July8-3_c.jpg"> | |
<br> | <br> | ||
<br> | <br> | ||
Line 2,954: | Line 3,599: | ||
<!--Main Culture--> | <!--Main Culture--> | ||
<strong>Main Culture</strong> | <strong>Main Culture</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 2,962: | Line 3,608: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>d</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>h</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>l</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 2,980: | Line 3,626: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Main Culture--> | <!--Main Culture--> | ||
Line 2,986: | Line 3,633: | ||
<!--Mini Prep--> | <!--Mini Prep--> | ||
<strong>Mini Prep</strong> | <strong>Mini Prep</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 3,012: | Line 3,660: | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Mini Prep--> | <!--Mini Prep--> | ||
Line 3,018: | Line 3,667: | ||
<!--Restriction Digest--> | <!--Restriction Digest--> | ||
<strong>Restriction Digest</strong> | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 3,032: | Line 3,682: | ||
<td>p427-TEF(d)</td> | <td>p427-TEF(d)</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
Line 3,040: | Line 3,690: | ||
<td>p427-TEF(h)</td> | <td>p427-TEF(h)</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
Line 3,048: | Line 3,698: | ||
<td>p427-TEF(l)</td> | <td>p427-TEF(l)</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
Line 3,055: | Line 3,705: | ||
<td>p427-TEF(ア)</td> | <td>p427-TEF(ア)</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
Line 3,063: | Line 3,713: | ||
<td>p427-TEF(イ)</td> | <td>p427-TEF(イ)</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
Line 3,071: | Line 3,721: | ||
<td>p427-TEF(エ)</td> | <td>p427-TEF(エ)</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
<td>Fast digest buffer</td> | <td>Fast digest buffer</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
<!--Restriction Digest--> | <!--Restriction Digest--> | ||
Line 3,082: | Line 3,733: | ||
<!--AGE--> | <!--AGE--> | ||
<strong>AGE</strong> | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
Line 3,095: | Line 3,747: | ||
<tr> | <tr> | ||
<td>1</td> | <td>1</td> | ||
- | <td> | + | <td>p427-TEF(ア)</td> |
- | <td | + | <td>CuMTS3</td> |
+ | <td>pGEX6P-2F1, pGEX6P-2R</td> | ||
+ | <td>Non Treated</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 3,102: | Line 3,756: | ||
<td>p427-TEF(ア)</td> | <td>p427-TEF(ア)</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>3</td> | <td>3</td> | ||
- | <td>p427-TEF( | + | <td>p427-TEF(イ)</td> |
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td>Non Treated</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 3,116: | Line 3,770: | ||
<td>p427-TEF(イ)</td> | <td>p427-TEF(イ)</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>5</td> | <td>5</td> | ||
- | <td>p427-TEF( | + | <td>p427-TEF(エ)</td> |
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td>Non Treated</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 3,130: | Line 3,784: | ||
<td>p427-TEF(エ)</td> | <td>p427-TEF(エ)</td> | ||
<td>CuMTS3</td> | <td>CuMTS3</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>7</td> | <td>7</td> | ||
- | <td>p427-TEF( | + | <td>p427-TEF(d)</td> |
- | <td> | + | <td>CuMTS62</td> |
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td>Non Treated</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 3,144: | Line 3,798: | ||
<td>p427-TEF(d)</td> | <td>p427-TEF(d)</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>9</td> | <td>9</td> | ||
- | <td>p427-TEF( | + | <td>p427-TEF(h)</td> |
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td>Non Treated</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 3,158: | Line 3,812: | ||
<td>p427-TEF(h)</td> | <td>p427-TEF(h)</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>11</td> | <td>11</td> | ||
- | <td>p427-TEF( | + | <td>p427-TEF(l)</td> |
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td>Non Treated</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 3,172: | Line 3,826: | ||
<td>p427-TEF(l)</td> | <td>p427-TEF(l)</td> | ||
<td>CuMTS62</td> | <td>CuMTS62</td> | ||
- | <td> | + | <td>pGEX6P-2F1, pGEX6P-2R</td> |
- | <td> | + | <td><em>Spe</em>1, <em>Xho</em>1</td> |
- | </ | + | |
- | < | + | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | </div> | ||
Marker: 1kb Ladder | Marker: 1kb Ladder | ||
<br> | <br> | ||
+ | <img src="/wiki/images/5/59/Kit_July9_c.jpg"> | ||
<!--AGE--> | <!--AGE--> | ||
<br> | <br> | ||
- | </p> | + | </p> |
- | <h2>August</h2> | + | </div> |
+ | </div> | ||
+ | |||
+ | <div class="accordion"> | ||
+ | <h2><a href="javascript:void(0);">August</a></h2> | ||
<div class="scroll"></div> | <div class="scroll"></div> | ||
+ | <div class="ac"> | ||
+ | <h3>August 19, 2014</h3> | ||
<p class="sentence"> | <p class="sentence"> | ||
+ | KOBAYASHI | ||
<br> | <br> | ||
+ | Conduct mutagenesis to remove an illegal restriction site (<em>Eco</em>R1) | ||
<br> | <br> | ||
- | < | + | <strong>InversePCR</strong> |
- | < | + | <div class="m_table"> |
- | < | + | <table class="materials"> |
- | < | + | <tr> |
- | < | + | <th colspan="4">Sample</th> |
- | < | + | </tr> |
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | <th>DNA polymerase</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>Terpinene3-mutagenesis-FP, Terpinene3-mutagenesis-RP</td> | ||
+ | <td>KOD-Plus</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | (TOYOBO, KOD-Plus-Mutagenesis Kit) | ||
+ | Start from KOD -Plus- Mutagenesis Kit protocol No.2. | ||
<br> | <br> | ||
- | |||
- | |||
- | |||
- | |||
- | |||
<br> | <br> | ||
- | < | + | </p> |
+ | |||
+ | |||
+ | <h3>August 20, 2014</h3> | ||
+ | <p class="sentence"> | ||
+ | KOBAYASHI | ||
<br> | <br> | ||
- | + | Restart mutagenesis from KOD -Plus- Mutagenesis Kit protocol No.3 to obtain plasmid (muta-CuMTS3 in pGEX-6P-2) | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
<br> | <br> | ||
- | < | + | <strong>Transformation</strong> |
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="4">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td rowspan="2">DH5α</td> | ||
+ | <td rowspan="2">pGEX6P-2</td> | ||
+ | <td rowspan="2">muta-CuMTS3</td> | ||
+ | <td>Terpinene3-mutagenesis-FP and Terpinene3-mutagenesis-RP</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
<br> | <br> | ||
- | |||
- | |||
- | |||
- | |||
- | |||
<br> | <br> | ||
- | < | + | </p> |
+ | |||
+ | <h3>August 21, 2014</h3> | ||
+ | <p class="sentence"> | ||
+ | KOBAYASHI | ||
<br> | <br> | ||
- | <br> | + | <strong>Colony Isolation</strong><br> |
- | < | + | Isolate each colony from two plates which were inoculated overnight |
- | < | + | <div class="m_table"> |
- | < | + | <table class="materials"> |
- | <br> | + | <tr> |
+ | <th colspan="4">Sample</th> | ||
+ | <th rowspan="2">Section</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>Terpinene3-mutagenesis-FP,<br>Terpinene3-mutagenesis-RP</td> | ||
+ | <td>あ-み</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | Select 5 isolated colonies whose code names are あ,か,さ,た and な and miniprep them. | ||
<br> | <br> | ||
<br> | <br> | ||
+ | </p> | ||
+ | |||
+ | <h3>August 22, 2014</h3> | ||
+ | <p class="sentence"> | ||
+ | KOBAYASHI | ||
<br> | <br> | ||
- | <br></p> | + | <strong>Restriction Digest</strong> |
- | <h2>September</h2> | + | <div class="m_table"> |
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | <th rowspan="2">Buffer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α あ</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Sal</em>1</td> | ||
+ | <td>Fast Digest buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α か</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Sal</em>1</td> | ||
+ | <td>Fast Digest buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α さ</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Sal</em>1</td> | ||
+ | <td>Fast Digest buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α た</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Sal</em>1</td> | ||
+ | <td>Fast Digest buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="4">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Marker</th> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td></td> | ||
+ | <td>DH5α あ</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Sal</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td></td> | ||
+ | <td>DH5α か</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Sal</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td></td> | ||
+ | <td>DH5α さ</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Sal</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>5</td> | ||
+ | <td></td> | ||
+ | <td>DH5α た</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Sal</em>1</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | <th rowspan="2">Buffer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α か</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | <td>Fast Digest buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α か</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | <td>Fast Digest buffer</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="4">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Marker</th> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td></td> | ||
+ | <td>DH5α か</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td></td> | ||
+ | <td>DH5α か</td> | ||
+ | <td>pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <img src="/wiki/images/b/bc/Kit_August22_c.jpg"> | ||
+ | An illegal <em>Eco</em>R1 restriction site was removed.<br> | ||
+ | <br> | ||
+ | <strong>PCR</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">DNA polymerase</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>か pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix and terpinene3-suffix</td> | ||
+ | <td>KOD-FX-NEO-Plus</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="4">Sample</th> | ||
+ | <th rowspan="2">DNA polymerase</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Marker</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td></td> | ||
+ | <td>か pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix and terpinene3-suffix</td> | ||
+ | <td>KOD-FX-NEO-Plus</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <img src="/wiki/images/7/72/Kit_August22_1_c.jpg"> | ||
+ | <br> | ||
+ | <br> | ||
+ | </p> | ||
+ | |||
+ | <h3>August 30, 2014</h3> | ||
+ | KOBAYASHI | ||
+ | <br> | ||
+ | <strong>PCR</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">DNA polymerase</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>か pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix-<em>Xba</em>1 and terpinene3-suffix-<em>Spe</em>1</td> | ||
+ | <td><em>Taq</em></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="4">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Marker</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td></td> | ||
+ | <td>か pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix-<em>Xba</em>1 and terpinene3-suffix-<em>Spe</em>1</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <img src="/wiki/images/3/31/Kit_August30_c.jpg"> | ||
+ | Made total amount of 295 μL of PCR samples. | ||
+ | <br> | ||
+ | </tr> | ||
+ | <br> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="accordion"> | ||
+ | <h2><a href="javascript:void(0);">September</a></h2> | ||
<div class="scroll"></div> | <div class="scroll"></div> | ||
+ | <div class="ac"> | ||
<p class="sentence"> | <p class="sentence"> | ||
+ | <h3>September 18, 2014</h3> | ||
+ | KOBAYASHI | ||
<br> | <br> | ||
+ | <strong>Miniprep</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
<br> | <br> | ||
- | < | + | <strong>Restriction Digest</strong> |
- | < | + | <div class="m_table"> |
- | < | + | <table class="materials"> |
- | < | + | <tr> |
- | < | + | <th colspan="3">Sample</th> |
- | < | + | <th rowspan="2">Buffer</th> |
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td><em>Xba</em>1, <em>Spe</em>1</td> | ||
+ | <td>Fast Digest buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | </tr> | ||
<br> | <br> | ||
- | < | + | <strong>AGE</strong> |
- | < | + | <div class="m_table"> |
- | < | + | <table class="materials"> |
- | < | + | <tr> |
- | < | + | <th rowspan="2">Lane</th> |
+ | <th colspan="4">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Marker</th> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td></td> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td></td> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td><em>Xba</em>1, <em>Spe</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td></td> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <img src="/wiki/images/2/29/Kit_September18_c.jpg"> | ||
<br> | <br> | ||
- | < | + | <strong>PCR</strong> |
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">DNA polymerase</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>か pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix-<em>Xba</em>1 and terpinene3-suffix-<em>Spe</em>1</td> | ||
+ | <td><em>Taq</em></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
<br> | <br> | ||
- | < | + | <strong>DNA Refinement 1</strong> |
- | < | + | <div class="m_table"> |
- | < | + | <table class="materials"> |
- | < | + | <tr> |
- | < | + | <th colspan="2">Sample</th> |
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix-<em>Xba</em>1 and terpinene3-suffix-<em>Spe</em>1</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | </tr> | ||
<br> | <br> | ||
- | < | + | <strong>Restriction Digest</strong> |
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | <th rowspan="2">Buffer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1, <em>Spe</em>1</td> | ||
+ | <td>Fast Digest</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>か pGEX6P-2</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix-<em>Xba</em>1 and terpinene3-suffix-<em>Spe</em>1</td> | ||
+ | <td><em>Xba</em>1, <em>Spe</em>1</td> | ||
+ | <td>Fast Digest</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | </tr> | ||
<br> | <br> | ||
- | < | + | <strong>DNA Refinement 2</strong> |
- | < | + | <div class="m_table"> |
- | < | + | <table class="materials"> |
- | < | + | <tr> |
- | < | + | <th colspan="2">Sample</th> |
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix-<em>Xba</em>1 and terpinene3-suffix-<em>Spe</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | </tr> | ||
<br> | <br> | ||
- | < | + | <strong>Ligation</strong> |
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>DNA</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix-<em>Xba</em>1 and terpinene3-suffix-<em>Spe<em>1</td> | ||
+ | <td><em>Spe</em>1, <em>Xba</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | </tr> | ||
+ | DNA ligase: DNA Ligation Kit ‹Mighty Mix› (TAKARA BIO INC.) | ||
<br> | <br> | ||
- | |||
- | |||
- | |||
- | |||
- | |||
<br> | <br> | ||
+ | <strong>Transformation</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="4">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix-<em>Xba</em>1 and terpinene3-suffix-<em>Spe</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | </tr> | ||
<br> | <br> | ||
+ | </p> | ||
+ | |||
+ | |||
+ | <h3>September 19, 2014</h3> | ||
+ | KOBAYASHI | ||
+ | <br> | ||
+ | <strong>Colony Isolation</strong> | ||
+ | <br> | ||
+ | Isolate each colony from the plates which were inoculated overnight. | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="4">Sample</th> | ||
+ | <th rowspan="2">Section</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix-<em>Xba</em>1 and terpinene3-suffix-<em>Spe</em>1</td> | ||
+ | <td>A-P</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>terpinene3-prefix-<em>Xba</em>1 and terpinene3-suffix-<em>Spe</em>1</td> | ||
+ | <td>ア-タ</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Colony Sweep</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td>ア</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td>イ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td>ウ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>5</td> | ||
+ | <td>エ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>6</td> | ||
+ | <td>オ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>7</td> | ||
+ | <td>カ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>8</td> | ||
+ | <td>キ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>9</td> | ||
+ | <td>ク</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>10</td> | ||
+ | <td>ケ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>11</td> | ||
+ | <td>コ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>12</td> | ||
+ | <td>サ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>13</td> | ||
+ | <td>シ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>14</td> | ||
+ | <td>ス</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>15</td> | ||
+ | <td>セ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>16</td> | ||
+ | <td>ソ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>17</td> | ||
+ | <td>タ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <img src="/wiki/images/4/40/Kit_September19_c.jpg"> | ||
+ | <br> | ||
+ | Targeted colonies were identified in セ and ソ. | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Colony Sweep</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td>A</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td>B</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td>C</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>5</td> | ||
+ | <td>D</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>6</td> | ||
+ | <td>E</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>7</td> | ||
+ | <td>F</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>8</td> | ||
+ | <td>G</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>9</td> | ||
+ | <td>H</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>10</td> | ||
+ | <td>I</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>11</td> | ||
+ | <td>J</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>12</td> | ||
+ | <td>K</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>13</td> | ||
+ | <td>L</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>14</td> | ||
+ | <td>M</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>15</td> | ||
+ | <td>N</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>16</td> | ||
+ | <td>O</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>17</td> | ||
+ | <td>P</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | </tr> | ||
+ | |||
+ | <img src="/wiki/images/e/e6/Kit_September19_1_c.jpg"> | ||
+ | <br> | ||
+ | Targeted colonies were identified in colony N and F. | ||
+ | <br> | ||
+ | <br> | ||
+ | Select セ, ソ, N, F | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Main Culture</strong> | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="4">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(N)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>----------</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>----------</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(セ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>----------</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(ソ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>----------</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | Cultivate these samples in 3mL of LB medium with ChlPhe at 30°C overnight. | ||
+ | <br> | ||
+ | </p> | ||
+ | |||
+ | |||
+ | <h3>September 20, 2014</h3> | ||
+ | KOBAYASHI | ||
+ | <br> | ||
+ | <strong>Miniprep</strong> | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(N)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(セ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(ソ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | Enzyme: E/P and <em>Xba</em>1 | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="4">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | <th rowspan="2">Buffer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Marker</th> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (セ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (ソ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (N)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (セ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (ソ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (N)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (セ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (ソ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>DH5α (N)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="4">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Marker</th> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (セ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (ソ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>5</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (N)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>6</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>7</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (セ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>8</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (ソ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>9</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>10</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (N)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>11</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>12</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>13</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>14</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (セ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>15</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (ソ)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>16</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>17</td> | ||
+ | <td></td> | ||
+ | <td>DH5α (N)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <img src="/wiki/images/e/e5/Kit_September20_c.jpg"> | ||
+ | <div class="clear"><hr /></div> | ||
+ | Sample F was selected as a submission plasmid. | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Main Culture</strong> | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="4">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>----------</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>----------</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td>----------</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | Cultivate these samples in 3mL of LB medium with ChlPhe at 30°C overnight. | ||
+ | <br> | ||
+ | </p> | ||
+ | |||
+ | |||
+ | <h3>September 21, 2014</h3> | ||
+ | KOBAYASHI | ||
+ | <br> | ||
+ | <strong>Miniprep</strong> | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="4">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α(F)</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="5">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | <th rowspan="2">Buffer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Marker</th> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1, <em>Xba</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>λDNA-HindⅢ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1, <em>Xba</em>1</td> | ||
+ | <td>FD buffer</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="5">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Marker</th> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>5</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>6</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>7</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1, <em>Xba</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>8</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>BBa_J04450</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>9</td> | ||
+ | <td>λDNA-HindⅢ</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>10</td> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>11</td> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>12</td> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>13</td> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>14</td> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>15</td> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td><em>Spe</em>1, <em>Xba</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>16</td> | ||
+ | <td></td> | ||
+ | <td>F</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>muta-CuMTS3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <img src="/wiki/images/d/d7/Kit_September21_c.jpg"> | ||
+ | |||
<br> | <br> | ||
<br></p> | <br></p> | ||
- | <h2>October</h2> | + | </div> |
+ | </div> | ||
+ | |||
+ | <div class="accordion"> | ||
+ | <h2><a href="javascript:void(0);">October</a></h2> | ||
<div class="scroll"></div> | <div class="scroll"></div> | ||
+ | <div class="ac"> | ||
<p class="sentence"> | <p class="sentence"> | ||
+ | |||
+ | <h3>October 1, 2014</h3> | ||
+ | KOBAYASHI | ||
<br> | <br> | ||
+ | <strong>Protein Extraction (<em>S. cerevisiae</em>)</strong> | ||
<br> | <br> | ||
- | < | + | <div class="m_table"> |
- | < | + | <table class="materials"> |
- | < | + | <tr> |
- | < | + | <th colspan="4">Sample</th> |
- | < | + | </tr> |
- | < | + | <tr> |
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE2</td> | ||
+ | <td>pGEX6P-2F1, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS62</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
<br> | <br> | ||
- | |||
- | |||
- | |||
- | |||
- | |||
<br> | <br> | ||
- | < | + | <strong>SDS-PAGE</strong> |
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="4">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE2</td> | ||
+ | <td>pGEX6P-2F1, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS62</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
<br> | <br> | ||
- | < | + | </tr> |
- | < | + | </p> |
- | < | + | |
- | < | + | |
- | + | <h3>October 2, 2014</h3> | |
+ | KOBAYASHI | ||
<br> | <br> | ||
- | < | + | <strong>Preculture</strong> |
<br> | <br> | ||
- | < | + | <div class="m_table"> |
- | < | + | <table class="materials"> |
- | < | + | <tr> |
- | < | + | <th colspan="4">Sample</th> |
- | < | + | </tr> |
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
<br> | <br> | ||
- | + | Cultivate these samples in 3mL of SD medium with G418 at 30°C overnight. | |
<br> | <br> | ||
- | |||
- | |||
- | |||
- | |||
- | |||
<br> | <br> | ||
+ | <strong>Western blot</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th colspan="4">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE2</td> | ||
+ | <td>pGEX6P-2F1, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS62</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
<br> | <br> | ||
+ | <img src="/wiki/images/e/e0/Kit_October2_c.jpg"> | ||
+ | </p> | ||
+ | |||
+ | <h3>October 3, 2014</h3> | ||
+ | KOBAYASHI | ||
<br> | <br> | ||
- | <br></p> | + | <strong>Main culture</strong> |
+ | <br> | ||
+ | Cultivate following samples in Big Scale with G418 at 28°C overnight in shaken culture. | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="4">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Host</th> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Restriction Digest</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | <th rowspan="2">Buffer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Spe</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1, <em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Linearized pSB1A3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | <td>Buffer H</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>AGE</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Lane</th> | ||
+ | <th rowspan="2">Marker</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>1kb Ladder</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Spe</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td></td> | ||
+ | <td>Linearized pSB1A3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>DNA Refinement 1</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Spe</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Xba</em>1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Linearized pSB1A3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1, <em>Dpn</em>1</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Ligation</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Vector</th> | ||
+ | <th colspan="2">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Insert1</th> | ||
+ | <th>Insert2</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1A3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Xba</em>1, <em>Spe</em>1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>pSB1A3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Xba</em>1, <em>Spe</em>1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | DNA ligase: DNA Ligation Kit ‹Mighty Mix› (TAKARA BIO INC.) | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Transformation</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert1</th> | ||
+ | <th>Insert2</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1A3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1A3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | </tr> | ||
+ | </p> | ||
+ | |||
+ | |||
+ | <h3>October 4, 2014</h3> | ||
+ | KOBAYASHI | ||
+ | <br> | ||
+ | <strong>Colony Isolation</strong> | ||
+ | <br> | ||
+ | Isolate each colony from the plates which were inoculated overnight. | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | <th rowspan="2">Section</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1A3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>1-16</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1A3</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | <td>a-p</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Monoterpene Extraction</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | Samples were stocked in -4℃. | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Preculture</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTSE2</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS62</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | Cultivate samples in 3mL of SD medium with G418 at 30°C overnight. | ||
+ | <br> | ||
+ | </tr> | ||
+ | </p> | ||
+ | |||
+ | |||
+ | <h3>October 5, 2014</h3> | ||
+ | KOBAYASHI | ||
+ | <br> | ||
+ | <strong>Miniprep</strong> | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="2">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1A3</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1A3</td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <br> | ||
+ | <strong>Restriction Digest and AGE</strong> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="2">Sample</th> | ||
+ | <th rowspan="2">Enzyme</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>DH5α</td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td></td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td></td> | ||
+ | <td>pSB1C3</td> | ||
+ | <td>K1448000</td> | ||
+ | <td><em>Eco</em>R1, <em>Pst</em>1</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | </tr> | ||
+ | </p> | ||
+ | |||
+ | <h3>October 8, 2014</h3> | ||
+ | KOBAYASHI | ||
+ | <br> | ||
+ | <strong>GC-MS analysis</strong> | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <strong>Preculture</strong> | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | |||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | </tr> | ||
+ | </p> | ||
+ | |||
+ | |||
+ | <h3>October 9, 2014</h3> | ||
+ | KOBAYASHI | ||
+ | <br> | ||
+ | <strong>Main Culture</strong> | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | </tr> | ||
+ | </p> | ||
+ | |||
+ | |||
+ | <h3>October 10, 2014</h3> | ||
+ | KOBAYASHI | ||
+ | <br> | ||
+ | <strong>GC Analysis</strong> | ||
+ | <br> | ||
+ | <div class="m_table"> | ||
+ | <table class="materials"> | ||
+ | <tr> | ||
+ | <th rowspan="2">Host</th> | ||
+ | <th colspan="3">Sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Vector</th> | ||
+ | <th>Insert</th> | ||
+ | <th>Primer</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td></td> | ||
+ | <td></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BY4247</td> | ||
+ | <td>p427-TEF</td> | ||
+ | <td>CuMTS3</td> | ||
+ | <td>pGEX6P-2F2, pGEX6P-2R</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
+ | <img src="/wiki/images/b/b1/Kit_October10_c.jpg"> | ||
+ | <div class="clear"><hr /></div> | ||
+ | For More Information: <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1448000">BBa_K1448000</a> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
<div class="clear"><hr /></div> | <div class="clear"><hr /></div> | ||
- | + | <div id='kitfooter-box' class='noprint'> | |
<div><a href="http://www.mediawiki.org/" style="float:left;margin:10px auto;"><img src="/wiki/skins/common/images/poweredby_mediawiki_88x31.png" height="31" width="88" alt="Powered by MediaWiki" /></a> | <div><a href="http://www.mediawiki.org/" style="float:left;margin:10px auto;"><img src="/wiki/skins/common/images/poweredby_mediawiki_88x31.png" height="31" width="88" alt="Powered by MediaWiki" /></a> | ||
- | </div> | + | </div> |
+ | <div><a href="http://creativecommons.org/licenses/by/3.0/" style="float:right;margin:10px auto;z-index:0;"><img src="http://i.creativecommons.org/l/by/3.0/88x31.png" alt="Attribution 3.0 Unported" width="88" height="31" /></a> | ||
<div id="kitfooter"> | <div id="kitfooter"> | ||
<ul id="fkit"> | <ul id="fkit"> | ||
Line 3,349: | Line 6,304: | ||
</ul> | </ul> | ||
</div> | </div> | ||
- | + | ||
</div> | </div> | ||
</div> <!-- close kitfooter-box --> | </div> <!-- close kitfooter-box --> | ||
Line 3,362: | Line 6,317: | ||
} | } | ||
- | th,td{ | + | th{ |
+ | border:1px solid rgba(300,300,300,300.5); | ||
+ | } | ||
+ | |||
+ | td{ | ||
border:1px solid rgba(300,300,300,300.5); | border:1px solid rgba(300,300,300,300.5); | ||
} | } | ||
Line 3,370: | Line 6,329: | ||
} | } | ||
+ | img{ | ||
+ | margin:10px 0px; | ||
+ | } | ||
</style> | </style> | ||
</html> | </html> | ||
{{Team:KIT-Kyoto/mobile}} | {{Team:KIT-Kyoto/mobile}} | ||
{{Team:KIT-Kyoto/footer}} | {{Team:KIT-Kyoto/footer}} |
Latest revision as of 01:13, 18 October 2014
LabNote
June
June 10, 2014
OKAMOTO
Miniprep
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3) | pGEX6P-2 | CuMTSE1 |
BL21(DE3) | pGEX6P-2 | CuMTSE2 |
BL21(DE3) | pGEX6P-2 | CuMTS3 |
BL21(DE3) | pGEX6P-2 | CuMTS62 |
Restriction Digest
Host | Sample | Enzyme | Buffer | |
---|---|---|---|---|
Vector | Insert | |||
BL21(DE3) | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTSE1 | Sal1, Not1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTSE2 | Sal1, Not1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTS3 | Sal1, Not1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTS62 | Sal1, Not1 | Buffer H |
June 11, 2014
KOBAYASHI
AGE
Lane | Sample | Enzyme | ||
---|---|---|---|---|
Marker | Vector | Insert | ||
1 | λDNA-HindⅢ | |||
2 | pGEX6P-2 | CuMTSE1 | Non Treated | |
3 | pGEX6P-2 | CuMTSE1 | EcoR1 | |
4 | pGEX6P-2 | CuMTSE1 | Sal1, Not1 | |
5 | pGEX6P-2 | CuMTSE2 | Non Treated | |
6 | pGEX6P-2 | CuMTSE2 | EcoR1 | |
7 | pGEX6P-2 | CuMTSE2 | Sal1, Not1 | |
8 | pGEX6P-2 | CuMTS3 | Non Treated | |
9 | pGEX6P-2 | CuMTS3 | EcoR1 | |
10 | pGEX6P-2 | CuMTS3 | Sal1, Not1 | |
11 | pGEX6P-2 | CuMTS62 | Non Treated | |
12 | pGEX6P-2 | CuMTS62 | EcoR1 | |
13 | pGEX6P-2 | CuMTS62 | Sal1, Not1 |
June 12, 2014
KOBAYASHI
Transformation
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 |
June 13, 2014
KOBAYASHI
Colony Isolation
Select 6 colonies per plate from 4 plates from June 12.
Host | Sample | Section | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | 1-a, 1-b |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | 2-a, 2-b |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 | 3-a, 3-b |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 | 6-a, 6-b |
June 14, 2014
ONISHI
Miniprep
Host | Sample | |
---|---|---|
Vector | Insert | |
1-a | pGEX6P-2 | CuMTSE1 |
1-b | pGEX6P-2 | CuMTSE1 |
2-a | pGEX6P-2 | CuMTSE2 |
2-b | pGEX6P-2 | CuMTSE2 |
3-a | pGEX6P-2 | CuMTS3 |
3-b | pGEX6P-2 | CuMTS3 |
6-a | pGEX6P-2 | CuMTS62 |
6-b | pGEX6P-2 | CuMTS62 |
Miniprep
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3) | pGEX6P-2 | CuMTSE1 |
BL21(DE3) | pGEX6P-2 | CuMTSE2 |
BL21(DE3) | pGEX6P-2 | CuMTS3 |
BL21(DE3) | pGEX6P-2 | CuMTS62 |
Restriction Digest
Host | Sample | Enzyme | Buffer | |
---|---|---|---|---|
Vector | Insert | |||
1-a | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
1-b | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
2-a | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
2-b | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
3-a | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
3-b | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
6-a | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
6-b | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
BL21(DE3) | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
AGE
Lane | Sample | Insert | Enzyme | Buffer | ||
---|---|---|---|---|---|---|
Marker | Host | Vector | ||||
1 | 1kb Ladder | |||||
2 | λDNA-HindⅢ | |||||
3 | 1-a | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H | |
4 | 1-b | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H | |
5 | 2-a | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H | |
6 | 2-b | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H | |
7 | 3-a | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H | |
8 | 3-b | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H | 9 | 6-a | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
10 | 6-b | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H | |
11 | BL21(DE3) | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H | |
12 | BL21(DE3) | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H | |
13 | BL21(DE3) | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H | |
14 | BL21(DE3) | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
Apply λDNA-HindⅢ to compare and contrast AGE of July 14 and June 11.
June 16, 2014
YAMAJI
Preculture
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 |
BL21(DE3)pLysS | pGEX6P-2 |
Main Culture
Host | Sample | IPTG | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | Induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | Non-induced |
June 18, 2014
YAMAJI
Preculture
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
Main Culture
Host | Sample | IPTG | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | Induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | Non-induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Non-induced |
Protein Extraction (E. coli)
Host | Sample | IPTG | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | Induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | Non-induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Non-induced |
Preparations for SDS-PAGE
Add SDS sample buffer to protein extractions in a flask, incubate at 37℃ for 15 minutes and keep it at 4℃.
June 19, 2014
KITOMI
SDS-PAGE
Host | Sample | IPTG | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | Induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | Non-induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Non-induced |
June 21, 2014
SOGA
Miniprep
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
PCR
Vector | Sample | DNA Polymerase | |
---|---|---|---|
Insert | Primer | ||
pGEX6P-2 | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R | KOD-FX-NEO |
AGE
Lane | Sample | Insert | Primer | ||
---|---|---|---|---|---|
Marker | Host | Vector | |||
1 | 1kb Ladder | ||||
2 | BY4247 | p427-TEF | |||
3 | pGEX6P-2 | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | ||
4 | pGEX6P-2 | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | ||
5 | pGEX6P-2 | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | ||
6 | pGEX6P-2 | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R |
June 24, 2014
SOGA
Preculture
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | ||
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
June 25, 2014
KOBAYASHI
Cultivate following samples in Big Scale at 30℃ overnight in shaken culture.
Host | Sample | |
---|---|---|
Vector | Insert | |
BL21(DE3)pLysS | ||
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
DNA Refinement 1
Sample | |
---|---|
DNA | Primer |
CuMTSE1 | pGEX6P-2F1, pGEX6P-2R |
CuMTSE1 | pGEX6P-2F2, pGEX6P-2R |
Restriction Digest
Vector | Sample | Enzyme | Buffer | |
---|---|---|---|---|
DNA | Primer | |||
CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Buffer H | |
CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Spe1, Xho1 | Buffer H | |
p427-TEF | Spe1, Xho1 | Buffer H |
DNA Refinement 2
Vector | Sample | Enzyme | |
---|---|---|---|
DNA | Primer | ||
CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | |
CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Spe1, Xho1 | |
p427-TEF | Spe1, Xho1 |
Main Culture
Host | Sample | IPTG | |
---|---|---|---|
Vector | Insert | ||
BL21(DE3)pLysS | pGEX6P-2 | Induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Induced |
BL21(DE3)pLysS | pGEX6P-2 | Non-induced | |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | Non-induced |
BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | Non-induced |
Ligation
Sample | ||
---|---|---|
Vector | DNA | Primer |
p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R |
p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R |
p427-TEF | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R |
p427-TEF | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R |
Transformation
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
DH5α | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R |
DH5α | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R |
DH5α | p427-TEF | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R |
DH5α | p427-TEF | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R |
June 26, 2014
SAWANO
Colony Isolation
Isolate each colony from the plates which were inoculated on June 25.
Host | Sample | Section | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
DH5α | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | 1~4 |
DH5α | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | a~h |
DH5α | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | ア~タ |
June 27, 2014
SHIMADA
Colony Sweep
Lane | Sample | ||
---|---|---|---|
Host | Vector | Insert | |
1 | BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
2 | 1 | ||
3 | 2 | ||
4 | 3 | ||
5 | 4 | ||
6 | a | ||
7 | b | ||
8 | c | ||
9 | d | ||
10 | e | ||
11 | f | ||
12 | g | ||
13 | h |
Colony Sweep
Lane | Sample | ||
---|---|---|---|
Host | Vector | Insert | |
1 | BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
2 | ア | ||
3 | イ | ||
4 | ウ | ||
5 | エ | ||
6 | オ | ||
7 | カ | ||
8 | キ | ||
9 | ク | ||
10 | ケ | ||
11 | コ | ||
12 | サ | ||
13 | シ | ||
14 | ス | ||
15 | セ | ||
16 | ソ | ||
17 | タ |
Identified targeted genes in 4, c, h, イ, カ, セ
Miniprep
Sample | |
---|---|
Host | |
4 | |
C | |
H | |
イ | |
カ | |
セ |
Restriction Digest
Sample | Enzyme | Buffer |
---|---|---|
Host | ||
4 | Spe1, Xho1 | Buffer H |
C | Spe1, Xho1 | Buffer H |
H | Spe1, Xho1 | Buffer H |
イ | Spe1, Xho1 | Buffer H |
カ | Spe1, Xho1 | Buffer H |
AGE
Lane | Sample | Primer | Enzyme | ||
---|---|---|---|---|---|
Marker | Host | Insert | |||
1 | 1kb Ladder | ||||
2 | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | |||
3 | 4 | Spe1, Xho1 | |||
4 | 4 | Spe1, Xho1 | |||
5 | 4 | Non Treated | |||
6 | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | |||
7 | c | Spe1, Xho1 | |||
8 | c | Spe1, Xho1 | |||
9 | c | Non Treated | |||
10 | h | Spe1, Xho1 | |||
11 | h | Spe1, Xho1 | |||
12 | h | Non Treated |
Transformation (electroportation)
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R |
BY4247 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R |
June 30, 2014
SOGA
Inoculation
Divide each of 2 plates into 8 sections and inoculate following samples on them.
Host | Sample | Section | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
BY4247 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | A~H |
BY4247 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | 一~ハ |
July
July 1, 2014
KOBAYASHI
Protein Extraction (S. cerevisiae)
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R |
BY4247 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R |
July 2, 2014
ONISHI
Miniprep
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | ||
BY4247 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R |
BY4247 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R |
Restriction Digest
Sample | Enzyme | Buffer | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
p427-TEF | CuMTSE1 | EcoR1 | Buffer H | |
p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | EcoR1 | Buffer H |
p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | EcoR1 | Buffer H |
p427-TEF | CuMTSE1 | Sac1, Kpn1 | Buffer L | |
p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Sac1, Kpn1 | Buffer L |
p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Sac1, Kpn1 | Buffer L |
AGE
Lane | Sample | Enzyme | |||
---|---|---|---|---|---|
Marker | Vector | Insert | Primer | ||
1 | p427-TEF | Non Treated | |||
2 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Non Treated | |
3 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Non Treated | |
4 | 1kb Ladder | ||||
5 | p427-TEF | EcoR1 | |||
6 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | EcoR1 | |
7 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | EcoR1 | |
8 | 1kb Ladder | ||||
9 | p427-TEF | Kpn1 | |||
10 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Kpn1 | |
11 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Kpn1 | |
12 | 1kb Ladder | ||||
13 | p427-TEF | Sac1 | |||
14 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Sac1 | |
15 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Sac1 | |
Restriction Digest
Sample | Enzyme | Buffer | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
p427-TEF | CuMTSE1 | EcoR1 | Buffer H | |
p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | EcoR1 | Buffer H |
p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | EcoR1 | Buffer H |
p427-TEF | CuMTSE1 | Sac1 | Buffer L | |
p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Sac1 | Buffer L |
p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Sac1 | Buffer L |
p427-TEF | CuMTSE1 | Kpn1 | Buffer L | |
p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Kpn1 | Buffer L |
p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Kpn1 | Buffer L |
p427-TEF | CuMTSE1 | Spe1, Xho1 | Buffer H | |
p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Buffer H |
p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Spe1, Xho1 | Buffer H |
AGE
Lane | Sample | Enzyme | |||
---|---|---|---|---|---|
Marker | Vector | Insert | Primer | ||
1 | 1kb Ladder | ||||
2 | p427-TEF | CuMTSE1 | EcoR1 | ||
3 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | EcoR1 | |
4 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | EcoR1 | |
5 | p427-TEF | CuMTSE1 | Kpn1 | ||
6 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Kpn1 | |
7 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Kpn1 | |
8 | p427-TEF | CuMTSE1 | Sac1 | ||
9 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Sac1 | |
10 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Sac1 | |
11 | p427-TEF | CuMTSE1 | Spe1, Xho1 | ||
12 | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | |
13 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R | Spe1, Xho1 |
July 3, 2014
OKAMOTO
PCR
Sample | DNA Polymerase | ||
---|---|---|---|
Vector | Insert | Primer | |
pGEX6P-2 | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTS3 | pGEX6P-2F2, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTS62 | pGEX6P-2F2, pGEX6P-2R | KOD-FX-NEO |
Protein Extraction (S. cerevisiae)
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | ||
A | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R |
六 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R |
July 4, 2014
SHIMADA
AGE
Lane | Sample/Marker | |||
---|---|---|---|---|
Marker | Vector | Insert | Primer | |
1 | 1kb Ladder | |||
2 | pGEX6P-2 | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | |
3 | pGEX6P-2 | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R | |
4 | pGEX6P-2 | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | |
5 | pGEX6P-2 | CuMTS3 | pGEX6P-2F2, pGEX6P-2R | |
6 | pGEX6P-2 | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | |
7 | pGEX6P-2 | CuMTS62 | pGEX6P-2F2, pGEX6P-2R |
PCR
Sample | DNA Polymerase | ||
---|---|---|---|
Vector | Insert | Primer | |
pGEX6P-2 | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTS3 | pGEX6P-2F2, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | KOD-FX-NEO |
pGEX6P-2 | CuMTS62 | pGEX6P-2F2, pGEX6P-2R | KOD-FX-NEO |
AGE
Lane | Sample | |||
---|---|---|---|---|
Marker | Vector | Insert | Primer | |
1 | 1kb Ladder | |||
2 | pGEX6P-2 | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | |
3 | pGEX6P-2 | 1kb Ladder+K83:N91 | pGEX6P-2F2, pGEX6P-2R | |
4 | pGEX6P-2 | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | |
5 | pGEX6P-2 | CuMTS3 | pGEX6P-2F2, pGEX6P-2R | |
6 | pGEX6P-2 | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | |
7 | pGEX6P-2 | CuMTS62 | pGEX6P-2F2, pGEX6P-2R |
July 5, 2014
KOBAYASHI
SDS-PAGE
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BL21(DE3)pLysS | pGEX6P-2 | ||
4 | pGEX6P-2 | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R |
C | pGEX6P-2 | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R |
BY4247 | p427-TEF | ||
A | p427-TEF | CuMTSE1 | pGEX6P-2F1, pGEX6P-2R |
六 | p427-TEF | CuMTSE1 | pGEX6P-2F2, pGEX6P-2R |
July 6, 2014
SOGA
DNA Refinement 1
Sample | |
---|---|
DNA | Primer |
CuMTSE2 | pGEX6P-2F1, pGEX6P-2R |
CuMTSE2 | pGEX6P-2F2, pGEX6P-2R |
CuMTS3 | pGEX6P-2F1, pGEX6P-2R |
CuMTS62 | pGEX6P-2F1, pGEX6P-2R |
Restriction Digest
Vector | Sample | Enzyme | Buffer | |
---|---|---|---|---|
DNA | Primer | |||
CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Fast digest buffer | |
CuMTSE2 | pGEX6P-2F2, pGEX6P-2R | Spe1, Xho1 | Fast digest buffer | |
CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Fast digest buffer | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Fast digest buffer | p427-TEF | Spe1, Xho1 | Fast digest buffer |
DNA Refinement 2
Sample | Enzyme | |
---|---|---|
DNA | Primer | |
CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
CuMTSE2 | pGEX6P-2F2, pGEX6P-2R | Spe1, Xho1 |
Ligation
Vector | Sample | Enzyme | |
---|---|---|---|
DNA | Primer | ||
p427-TEF | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
p427-TEF | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R | Spe1, Xho1 |
DNA ligase: DNA Ligation Kit ‹Mighty Mix› (TAKARA BIO INC.)
Transformation (E. coli)
Host | Sample | Enzyme | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
DH5α | p427-TEF | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
DH5α | p427-TEF | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R | Spe1, Xho1 |
DH5α | p427-TEF | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
DH5α | p427-TEF | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
July 7, 2014
KOBAYASHI
Colony Isolation
Isolate each colony from the plates which were inoculated on July 6.
Host | Sample | Section | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
DH5α | p427-TEF | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R | 1~16 |
DH5α | p427-TEF | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | ア~エ |
DH5α | p427-TEF | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | a~p |
Transformation (E. coli)
Host | Sample | Enzyme | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
DH5α | p427-TEF | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R | Spe1, Xho1 |
July 8, 2014
OKAMOTO
Colony Sweep
Lane | Sample | |
---|---|---|
Host | ||
1 | DH5α | p427-TEF |
2 | a | |
3 | b | |
4 | c | |
5 | d | |
6 | e | |
7 | f | |
8 | g | |
9 | h | |
10 | i | |
11 | j | |
12 | k | |
13 | l | |
14 | m | |
15 | n | |
16 | o | |
17 | p |
Colony Sweep
Lane | Sample | |
---|---|---|
Host | ||
1 | DH5α | p427-TEF |
2 | ア | |
3 | イ | |
4 | ウ | |
5 | エ |
Colony Sweep
Lane | Sample | |
---|---|---|
Host | ||
1 | DH5α | p427-TEF |
2 | 1 | |
3 | 2 | |
4 | 3 | |
5 | 4 | |
6 | 5 | |
7 | 6 | |
8 | 7 | |
9 | 8 | |
10 | 9 | |
11 | 10 | |
12 | 11 | |
13 | 12 | |
14 | 13 | |
15 | 14 | |
16 | 15 | |
17 | 16 |
July 9, 2014
OKAMOTO
Main Culture
Sample |
---|
Host |
d |
h |
l |
ア |
イ |
エ |
Mini Prep
Sample |
---|
Host |
D |
H |
L |
ア |
イ |
エ |
Restriction Digest
Vector | Sample | Enzyme | Buffer | |
---|---|---|---|---|
Insert | Primer | |||
p427-TEF(d) | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Fast digest buffer |
p427-TEF(h) | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Fast digest buffer |
p427-TEF(l) | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Fast digest buffer |
p427-TEF(ア) | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Fast digest buffer |
p427-TEF(イ) | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Fast digest buffer |
p427-TEF(エ) | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 | Fast digest buffer |
AGE
Lane | Sample | Enzyme | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
1 | p427-TEF(ア) | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Non Treated |
2 | p427-TEF(ア) | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
3 | p427-TEF(イ) | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Non Treated |
4 | p427-TEF(イ) | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
5 | p427-TEF(エ) | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Non Treated |
6 | p427-TEF(エ) | CuMTS3 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
7 | p427-TEF(d) | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Non Treated |
8 | p427-TEF(d) | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
9 | p427-TEF(h) | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Non Treated |
10 | p427-TEF(h) | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
11 | p427-TEF(l) | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Non Treated |
12 | p427-TEF(l) | CuMTS62 | pGEX6P-2F1, pGEX6P-2R | Spe1, Xho1 |
August
August 19, 2014
KOBAYASHI
Conduct mutagenesis to remove an illegal restriction site (EcoR1)
InversePCR
Sample | |||
---|---|---|---|
Vector | Insert | Primer | DNA polymerase |
pGEX6P-2 | CuMTS3 | Terpinene3-mutagenesis-FP, Terpinene3-mutagenesis-RP | KOD-Plus |
August 20, 2014
KOBAYASHI
Restart mutagenesis from KOD -Plus- Mutagenesis Kit protocol No.3 to obtain plasmid (muta-CuMTS3 in pGEX-6P-2)
Transformation
Sample | |||
---|---|---|---|
Host | Vector | Insert | Primer |
DH5α | pGEX6P-2 | muta-CuMTS3 | Terpinene3-mutagenesis-FP and Terpinene3-mutagenesis-RP |
August 21, 2014
KOBAYASHI
Colony Isolation
Isolate each colony from two plates which were inoculated overnight
Sample | Section | |||
---|---|---|---|---|
Host | Vector | Insert | Primer | |
DH5α | pSB1C3 | muta-CuMTS3 | Terpinene3-mutagenesis-FP, Terpinene3-mutagenesis-RP |
あ-み |
August 22, 2014
KOBAYASHI
Restriction Digest
Sample | Enzyme | Buffer | ||
---|---|---|---|---|
Host | Vector | Insert | ||
DH5α あ | pGEX6P-2 | muta-CuMTS3 | Sal1 | Fast Digest buffer |
DH5α か | pGEX6P-2 | muta-CuMTS3 | Sal1 | Fast Digest buffer |
DH5α さ | pGEX6P-2 | muta-CuMTS3 | Sal1 | Fast Digest buffer |
DH5α た | pGEX6P-2 | muta-CuMTS3 | Sal1 | Fast Digest buffer |
AGE
Lane | Sample | Enzyme | |||
---|---|---|---|---|---|
Marker | Host | Vector | Insert | ||
1 | 1kb Ladder | ||||
2 | DH5α あ | pGEX6P-2 | muta-CuMTS3 | Sal1 | |
3 | DH5α か | pGEX6P-2 | muta-CuMTS3 | Sal1 | |
4 | DH5α さ | pGEX6P-2 | muta-CuMTS3 | Sal1 | |
5 | DH5α た | pGEX6P-2 | muta-CuMTS3 | Sal1 |
Restriction Digest
Sample | Enzyme | Buffer | ||
---|---|---|---|---|
Host | Vector | Insert | ||
DH5α か | pGEX6P-2 | CuMTS3 | EcoR1 | Fast Digest buffer |
DH5α か | pGEX6P-2 | muta-CuMTS3 | EcoR1 | Fast Digest buffer |
AGE
Lane | Sample | Enzyme | |||
---|---|---|---|---|---|
Marker | Host | Vector | Insert | ||
1 | 1kb Ladder | ||||
2 | DH5α か | pGEX6P-2 | CuMTS3 | EcoR1 | |
3 | DH5α か | pGEX6P-2 | muta-CuMTS3 | EcoR1 |
PCR
Sample | DNA polymerase | ||
---|---|---|---|
Vector | Insert | Primer | |
か pGEX6P-2 | muta-CuMTS3 | terpinene3-prefix and terpinene3-suffix | KOD-FX-NEO-Plus |
AGE
Lane | Sample | DNA polymerase | |||
---|---|---|---|---|---|
Marker | Vector | Insert | Primer | ||
1 | 1kb Ladder | ||||
2 | か pGEX6P-2 | muta-CuMTS3 | terpinene3-prefix and terpinene3-suffix | KOD-FX-NEO-Plus |
August 30, 2014
KOBAYASHIPCR
Sample | DNA polymerase | ||
---|---|---|---|
Vector | Insert | Primer | |
か pGEX6P-2 | muta-CuMTS3 | terpinene3-prefix-Xba1 and terpinene3-suffix-Spe1 | Taq |
AGE
Lane | Sample | |||
---|---|---|---|---|
Marker | Vector | Insert | Primer | |
1 | 1kb Ladder | |||
2 | か pGEX6P-2 | muta-CuMTS3 | terpinene3-prefix-Xba1 and terpinene3-suffix-Spe1 |
Made total amount of 295 μL of PCR samples.
September
September 18, 2014
KOBAYASHIMiniprep
Sample | ||
---|---|---|
Host | Vector | Insert |
DH5α | pSB1C3 | BBa_J04450 |
DH5α | pSB1C3 | BBa_J04450 |
DH5α | pSB1C3 | BBa_J04450 |
Restriction Digest
Sample | Buffer | ||
---|---|---|---|
Vector | Insert | Enzyme | |
pSB1C3 | BBa_J04450 | EcoR1, Pst1 | Buffer H |
pSB1C3 | BBa_J04450 | Xba1, Spe1 | Fast Digest buffer |
pSB1C3 | BBa_J04450 |
AGE
Lane | Sample | Enzyme | |||
---|---|---|---|---|---|
Marker | Host | Vector | Insert | ||
1 | 1kb Ladder | ||||
2 | DH5α | pSB1C3 | BBa_J04450 | EcoR1, Pst1 | |
3 | DH5α | pSB1C3 | BBa_J04450 | Xba1, Spe1 | |
4 | DH5α | pSB1C3 | BBa_J04450 |
PCR
Sample | DNA polymerase | ||
---|---|---|---|
Vector | Insert | Primer | |
か pGEX6P-2 | muta-CuMTS3 | terpinene3-prefix-Xba1 and terpinene3-suffix-Spe1 | Taq |
DNA Refinement 1
Sample | |
---|---|
Insert | Primer |
muta-CuMTS3 | terpinene3-prefix-Xba1 and terpinene3-suffix-Spe1 |
Restriction Digest
Sample | Enzyme | Buffer | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
pSB1C3 | Xba1, Spe1 | Fast Digest | ||
か pGEX6P-2 | muta-CuMTS3 | terpinene3-prefix-Xba1 and terpinene3-suffix-Spe1 | Xba1, Spe1 | Fast Digest |
DNA Refinement 2
Sample | |
---|---|
Insert | Primer |
muta-CuMTS3 | terpinene3-prefix-Xba1 and terpinene3-suffix-Spe1 |
Ligation
Sample | Enzyme | ||
---|---|---|---|
Vector | DNA | Primer | |
pSB1C3 | muta-CuMTS3 | terpinene3-prefix-Xba1 and terpinene3-suffix-Spe1 | Spe1, Xba1 |
Transformation
Sample | |||
---|---|---|---|
Host | Vector | Insert | Primer |
DH5α | pSB1C3 | muta-CuMTS3 | terpinene3-prefix-Xba1 and terpinene3-suffix-Spe1 |
September 19, 2014
KOBAYASHIColony Isolation
Isolate each colony from the plates which were inoculated overnight.
Sample | Section | |||
---|---|---|---|---|
Host | Vector | Insert | Primer | |
DH5α | pSB1C3 | muta-CuMTS3 | terpinene3-prefix-Xba1 and terpinene3-suffix-Spe1 | A-P |
DH5α | pSB1C3 | muta-CuMTS3 | terpinene3-prefix-Xba1 and terpinene3-suffix-Spe1 | ア-タ |
Colony Sweep
Lane | Sample | ||
---|---|---|---|
Host | Vector | Insert | |
1 | DH5α | pSB1C3 | muta-CuMTS3 |
2 | ア | ||
3 | イ | ||
4 | ウ | ||
5 | エ | ||
6 | オ | ||
7 | カ | ||
8 | キ | ||
9 | ク | ||
10 | ケ | ||
11 | コ | ||
12 | サ | ||
13 | シ | ||
14 | ス | ||
15 | セ | ||
16 | ソ | ||
17 | タ |
Targeted colonies were identified in セ and ソ.
Colony Sweep
Lane | Sample | ||
---|---|---|---|
Host | Vector | Insert | |
1 | DH5α | pSB1C3 | muta-CuMTS3 |
2 | A | ||
3 | B | ||
4 | C | ||
5 | D | ||
6 | E | ||
7 | F | ||
8 | G | ||
9 | H | ||
10 | I | ||
11 | J | ||
12 | K | ||
13 | L | ||
14 | M | ||
15 | N | ||
16 | O | ||
17 | P |
Targeted colonies were identified in colony N and F.
Select セ, ソ, N, F
Main Culture
Sample | |||
---|---|---|---|
Host | Vector | Insert | Primer |
DH5α(N) | pSB1C3 | muta-CuMTS3 | ---------- |
DH5α(F) | pSB1C3 | muta-CuMTS3 | ---------- |
DH5α(セ) | pSB1C3 | muta-CuMTS3 | ---------- |
DH5α(ソ) | pSB1C3 | muta-CuMTS3 | ---------- |
Cultivate these samples in 3mL of LB medium with ChlPhe at 30°C overnight.
September 20, 2014
KOBAYASHIMiniprep
Sample | ||
---|---|---|
Host | Vector | Primer |
DH5α(N) | pSB1C3 | muta-CuMTS3 |
DH5α(F) | pSB1C3 | muta-CuMTS3 |
DH5α(セ) | pSB1C3 | muta-CuMTS3 |
DH5α(ソ) | pSB1C3 | muta-CuMTS3 |
Enzyme: E/P and Xba1
Restriction Digest
Sample | Enzyme | Buffer | |||
---|---|---|---|---|---|
Marker | Host | Vector | Insert | ||
pSB1C3 | |||||
DH5α (セ) | pSB1C3 | muta-CuMTS3 | |||
DH5α (ソ) | pSB1C3 | muta-CuMTS3 | |||
DH5α (F) | pSB1C3 | muta-CuMTS3 | |||
DH5α (N) | pSB1C3 | muta-CuMTS3 | |||
DH5α (セ) | pSB1C3 | EcoR1, Pst1 | Buffer H | ||
DH5α (ソ) | pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | Buffer H | |
DH5α (F) | pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | Buffer H | |
DH5α (N) | pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | Buffer H | |
pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | Buffer H | ||
pSB1C3 | Xba1 | FD buffer | |||
DH5α (セ) | pSB1C3 | muta-CuMTS3 | Xba1 | FD buffer | |
DH5α (ソ) | pSB1C3 | muta-CuMTS3 | Xba1 | FD buffer | |
DH5α (F) | pSB1C3 | muta-CuMTS3 | Xba1 | FD buffer | |
DH5α (N) | pSB1C3 | muta-CuMTS3 | Xba1 | FD buffer |
AGE
Lane | Sample | Enzyme | |||
---|---|---|---|---|---|
Marker | Host | Vector | Insert | ||
1 | pSB1C3 | ||||
2 | DH5α (セ) | pSB1C3 | muta-CuMTS3 | ||
3 | DH5α (ソ) | pSB1C3 | muta-CuMTS3 | ||
4 | DH5α (F) | pSB1C3 | muta-CuMTS3 | ||
5 | DH5α (N) | pSB1C3 | muta-CuMTS3 | ||
6 | 1kb Ladder | ||||
7 | DH5α (セ) | pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | |
8 | DH5α (ソ) | pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | |
9 | DH5α (F) | pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | |
10 | DH5α (N) | pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | |
11 | pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | ||
12 | 1kb Ladder | ||||
13 | pSB1C3 | Xba1 | |||
14 | DH5α (セ) | pSB1C3 | muta-CuMTS3 | Xba1 | |
15 | DH5α (ソ) | pSB1C3 | muta-CuMTS3 | Xba1 | |
16 | DH5α (F) | pSB1C3 | muta-CuMTS3 | Xba1 | |
17 | DH5α (N) | pSB1C3 | muta-CuMTS3 | Xba1 |
Main Culture
Sample | |||
---|---|---|---|
Host | Vector | Insert | Primer |
DH5α(F) | pSB1C3 | muta-CuMTS3 | ---------- |
DH5α(F) | pSB1C3 | muta-CuMTS3 | ---------- |
DH5α(F) | pSB1C3 | muta-CuMTS3 | ---------- |
Cultivate these samples in 3mL of LB medium with ChlPhe at 30°C overnight.
September 21, 2014
KOBAYASHIMiniprep
Sample | |||
---|---|---|---|
Host | Vector | Insert | Primer |
DH5α(F) | pSB1C3 | muta-CuMTS3 | |
DH5α(F) | pSB1C3 | muta-CuMTS3 | |
DH5α(F) | pSB1C3 | muta-CuMTS3 |
Restriction Digest
Sample | Enzyme | Buffer | ||||
---|---|---|---|---|---|---|
Marker | Host | Vector | Insert | Primer | ||
1kb Ladder | ||||||
pSB1C3 | BBa_J04450 | EcoR1 | Buffer H | |||
pSB1C3 | BBa_J04450 | Pst1 | Buffer H | |||
pSB1C3 | BBa_J04450 | EcoR1, Pst1 | Buffer H | |||
pSB1C3 | BBa_J04450 | Spe1 | FD buffer | |||
pSB1C3 | BBa_J04450 | Xba1 | FD buffer | |||
pSB1C3 | BBa_J04450 | Spe1, Xba1 | FD buffer | |||
pSB1C3 | BBa_J04450 | |||||
λDNA-HindⅢ | ||||||
F | pSB1C3 | muta-CuMTS3 | EcoR1 | Buffer H | ||
F | pSB1C3 | muta-CuMTS3 | Pst1 | Buffer H | ||
F | pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | Buffer H | ||
F | pSB1C3 | muta-CuMTS3 | Spe1 | FD buffer | ||
F | pSB1C3 | muta-CuMTS3 | Xba1 | FD buffer | ||
F | pSB1C3 | muta-CuMTS3 | Spe1, Xba1 | FD buffer | ||
F | pSB1C3 | muta-CuMTS3 |
AGE
Lane | Sample | Enzyme | ||||
---|---|---|---|---|---|---|
Marker | Host | Vector | Insert | Primer | ||
1 | 1kb Ladder | |||||
2 | pSB1C3 | BBa_J04450 | EcoR1 | |||
3 | pSB1C3 | BBa_J04450 | Pst1 | |||
4 | pSB1C3 | BBa_J04450 | EcoR1, Pst1 | |||
5 | pSB1C3 | BBa_J04450 | Spe1 | |||
6 | pSB1C3 | BBa_J04450 | Xba1 | |||
7 | pSB1C3 | BBa_J04450 | Spe1, Xba1 | |||
8 | pSB1C3 | BBa_J04450 | ||||
9 | λDNA-HindⅢ | |||||
10 | F | pSB1C3 | muta-CuMTS3 | EcoR1 | ||
11 | F | pSB1C3 | muta-CuMTS3 | Pst1 | ||
12 | F | pSB1C3 | muta-CuMTS3 | EcoR1, Pst1 | ||
13 | F | pSB1C3 | muta-CuMTS3 | Spe1 | ||
14 | F | pSB1C3 | muta-CuMTS3 | Xba1 | ||
15 | F | pSB1C3 | muta-CuMTS3 | Spe1, Xba1 | ||
16 | F | pSB1C3 | muta-CuMTS3 |
October
October 1, 2014
KOBAYASHIProtein Extraction (S. cerevisiae)
Sample | |||
---|---|---|---|
Host | Vector | Insert | Primer |
BY4247 | p427-TEF | ||
BY4247 | p427-TEF | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R |
BY4247 | p427-TEF | CuMTS3 | pGEX6P-2F2, pGEX6P-2R |
BY4247 | p427-TEF | CuMTS62 | pGEX6P-2F2, pGEX6P-2R |
SDS-PAGE
Lane | Sample | |||
---|---|---|---|---|
Host | Vector | Insert | Primer | |
1 | BY4247 | p427-TEF | ||
2 | BY4247 | p427-TEF | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R |
3 | BY4247 | p427-TEF | CuMTS3 | pGEX6P-2F2, pGEX6P-2R |
4 | BY4247 | p427-TEF | CuMTS62 | pGEX6P-2F2, pGEX6P-2R |
October 2, 2014
KOBAYASHIPreculture
Sample | |||
---|---|---|---|
Host | Vector | Insert | Primer |
BY4247 | p427-TEF | ||
BY4247 | p427-TEF | CuMTS3 | pGEX6P-2F2, pGEX6P-2R |
Cultivate these samples in 3mL of SD medium with G418 at 30°C overnight.
Western blot
Lane | Sample | |||
---|---|---|---|---|
Host | Vector | Insert | Primer | |
1 | BY4247 | p427-TEF | ||
2 | BY4247 | p427-TEF | CuMTSE2 | pGEX6P-2F1, pGEX6P-2R |
3 | BY4247 | p427-TEF | CuMTS3 | pGEX6P-2F2, pGEX6P-2R |
4 | BY4247 | p427-TEF | CuMTS62 | pGEX6P-2F2, pGEX6P-2R |
October 3, 2014
KOBAYASHIMain culture
Cultivate following samples in Big Scale with G418 at 28°C overnight in shaken culture.
Sample | |||
---|---|---|---|
Host | Vector | Insert | Primer |
BY4247 | p427-TEF | ||
BY4247 | p427-TEF | CuMTS3 | pGEX6P-2F2, pGEX6P-2R |
Restriction Digest
Sample | Enzyme | Buffer | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
pSB1C3 | EcoR1, Spe1 | Buffer H | ||
pSB1C3 | Xba1, Pst1 | Buffer H | ||
Linearized pSB1A3 | EcoR1, Pst1 | Buffer H |
AGE
Lane | Marker | Sample | Enzyme | ||
---|---|---|---|---|---|
Vector | Insert | Primer | |||
1 | 1kb Ladder | ||||
2 | pSB1C3 | EcoR1, Spe1 | |||
3 | pSB1C3 | Xba1, Pst1 | |||
4 | Linearized pSB1A3 | EcoR1, Pst1 |
DNA Refinement 1
Sample | Enzyme | ||
---|---|---|---|
Vector | Insert | Primer | |
pSB1C3 | EcoR1, Spe1 | ||
pSB1C3 | Xba1, Pst1 | ||
pSB1C3 | Xba1, Pst1 | ||
Linearized pSB1A3 | EcoR1, Pst1, Dpn1 |
Ligation
Vector | Sample | Enzyme | |
---|---|---|---|
Insert1 | Insert2 | ||
pSB1A3 | EcoR1, Xba1, Spe1, Pst1 | ||
pSB1A3 | EcoR1, Xba1, Spe1, Pst1 |
DNA ligase: DNA Ligation Kit ‹Mighty Mix› (TAKARA BIO INC.)
Transformation
Host | Sample | ||
---|---|---|---|
Vector | Insert1 | Insert2 | |
DH5α | pSB1A3 | ||
DH5α | pSB1A3 |
October 4, 2014
KOBAYASHIColony Isolation
Isolate each colony from the plates which were inoculated overnight.
Host | Sample | Section | ||
---|---|---|---|---|
Vector | Insert | Primer | ||
DH5α | pSB1A3 | 1-16 | ||
DH5α | pSB1A3 | a-p |
Monoterpene Extraction
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | ||
BY4247 | p427-TEF | CuMTS3 | pGEX6P-2F2, pGEX6P-2R |
Samples were stocked in -4℃.
Preculture
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | CuMTSE2 | pGEX6P-2F2, pGEX6P-2R |
BY4247 | p427-TEF | CuMTS62 | pGEX6P-2F2, pGEX6P-2R |
Cultivate samples in 3mL of SD medium with G418 at 30°C overnight.
October 5, 2014
KOBAYASHIMiniprep
Host | Sample | |
---|---|---|
Vector | Insert | |
DH5α | pSB1A3 | |
DH5α | pSB1A3 |
Restriction Digest and AGE
Host | Sample | Enzyme | |
---|---|---|---|
Vector | Insert | ||
DH5α | pSB1C3 | EcoR1, Pst1 | |
DH5α | pSB1C3 | EcoR1, Pst1 | |
pSB1C3 | K1448000 | EcoR1, Pst1 |
October 8, 2014
KOBAYASHIGC-MS analysis
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | ||
BY4247 | p427-TEF | CuMTS3 | pGEX6P-2F2, pGEX6P-2R |
Preculture
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | ||
BY4247 | p427-TEF | CuMTS3 | pGEX6P-2F2, pGEX6P-2R |
October 9, 2014
KOBAYASHIMain Culture
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | ||
BY4247 | p427-TEF | CuMTS3 | pGEX6P-2F2, pGEX6P-2R |
October 10, 2014
KOBAYASHIGC Analysis
Host | Sample | ||
---|---|---|---|
Vector | Insert | Primer | |
BY4247 | p427-TEF | ||
BY4247 | p427-TEF | CuMTS3 | pGEX6P-2F2, pGEX6P-2R |