Team:KIT-Kyoto/kazukokekokko

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{{KIT-Kyoto}}
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{{KIT-Kyoto.Header}}
<html>
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<head>
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<meta charset="UTF-8">
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<script type="text/javascript" src="http://ajax.googleapis.com/ajax/libs/jquery/1.4.2/jquery.min.js"></script>
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<div id="my_body">
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<div id="mynavi">
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<ul>
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<li>Takaaki TAKEDA</li>
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<li>Shunji MATSUZAKI</li>
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</ul>
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<div id="my_contents">
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<div id="mymain">
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<div class="contents"><h2>PROFILE</h2>
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<p></p></div>
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<!--まだ終わってないやつ--!>
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<font size="6">idea note</font>
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スクラップみたいにしてみる？？？？
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<div class="box1">
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<p><img src="img/img1.jpg" title="TAKAAKI" width="150" height="150" class="css-hover" /></p>
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<p><strong><u>Takaaki Takeda</u></strong></p>
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<p>detail<br />
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<br />
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</p>
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</div>
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<div class="box2">
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<p><img src="img/img1.jpg" title="Shunji" width="150" height="150" class="css-hover" /></p>
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<p><strong><u>Shunji Matsuzaki</u></strong></p>
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<p>detail<br />
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</p>
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</div>
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<style type="text/css">
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/*ボックス1の設定*/
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width: 730px; /* ボックスの幅 */
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height: 220px;
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<br>
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<body>
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<div id="HIDARI">
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<ul class="acc">
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  <li><a href="1"><img src="" width="160" height="32" class="menu_head" />LAB NOTE</a>  
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    <ul class="fxmn">  
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      <li><div><a href="">First</a></div></li>  
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      <li><div><a href="">Week1 ()</a></div></li>  
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      <li><div><a href="">Week2 ()</a></div></li>  
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      <li><div><a href="">Week3 ()</a></div></li>  
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      <li><div><a href="">Week4 ()</a></div></li>
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-
      <li><div><a href="">Week5 ()</a></div></li>  
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-
      <li><div><a href="">Week6 ()</a></div></li>  
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      <li><div><a href="">Week7 ()</a></div></li>  
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-
      <li><div><a href="">Week8 ()</a></div></li>
+
-
      <li><div><a href="">Week9 ()</a></div></li>  
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-
      <li><div><a href="">Week10 ()</a></div></li>  
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-
    </ul>  
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-
  </li>  
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-
</ul>
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-
<a href=""><img src="" width="160" height="32" border="0" bordercolor="black">MEETING</a>
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-
<BR>
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<a href=""><img src="" width="160" height="32" border="0" bordercolor="black">PROTOCOL</a>
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-
<BR>
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<a href="1"><img src="" width="160" height="32" class="menu_head" />DESIGN NOTE</a>  
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</div>
</div>
-
</body>
 
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<BR>
 
<br>
<br>
-
pfofile ex.<A href="https://2012.igem.org/Team:KIT-Kyoto/Team-Kazuko" >いちいち押すやつ</A>
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</td>
 +
<td width="800px" height="510px" valign="top">
 +
<div id="MIGI">
 +
<font color="#f5b1aa"><u>TNFAIP3</u></font>
<br>
<br>
-
見出し <a href="http://midashi-maker.v-colors.com/">go</a>
+
<font color="#f5b1aa"><u>PARTS</u></font>
<br>
<br>
-
<a href="http://www.colordic.org/w/"> 日本の伝統色</a>
+
<h2>BP reaction by Invitrogen gateway system</h2>
<br>
<br>
 +
1. PCR using primers containing the attB sequence.
 +
<br>
 +
2. Purify PCR product.
 +
<br>
 +
3. Add the following components to a 1.5 mL microcentrifuge tube at room temperature and mix:
 +
<br><br>
 +
<Table Border Cellspacing="0">
 +
<Tr><Td>attB PCR product</Td><Td>75 ng/reaction (1-7 μL)</Td></Tr>
 +
<Tr><Td>pDONR vector</Td><Td>150ng/reaction (1 μL)</Td></Tr>
 +
<Tr><Td>TE Buffer</Td><Td>to 8 μL</Td></Tr>
 +
</Table>
 +
<br>
 +
4. Vortex BP ClonaseII enzyme mix briefly. Add 1 - 2 μL to the components above and mix well by vortexing and spin them down.
 +
<br>
 +
5. Incubate reaction at 25˚C for more than 1 hour.
 +
<br>
 +
6. Add 1 μL of 2 μg/μL Proteinase K solution and vortex briefly.
 +
<br>
 +
7. Incubate at 37˚C for 10 minutes.
 +
<br>
 +
<br>
 +
<h2>LR reaction by Invitrogen gateway system</h2>
 +
<br>
 +
1. Add the following components to a 1.5 mL microcentrifuge tube at room temperature and mix:
 +
<br><br>
 +
<Table Border Cellspacing="0">
 +
<Tr><Td>Entry clones</Td><Td>50-150 ng/reaction (1-7 μL)</Td></Tr>
 +
<tr><td>Destination vector</td><td>150ng/reaction (1 μL)</td></tr>
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<Tr><Td>TE Buffer</Td><Td>to 8 − 9 μL</Td></Tr>
 +
</Table>
 +
<br>
 +
2. Vortex LR ClonaseII enzyme mix briefly. Add 1 – 2 μL, to the components above and mix well by vortexing and spin down.
 +
<br>
 +
3. Incubate reaction at 25˚C for 16 hours.
 +
<br>
 +
4. Add 1 μL of 2 μg/μL Proteinase K solution and vortex briefly.
 +
<br>
 +
<br>
 +
5. Incubate at 37˚C for 10 minutes.
 +
<br>
 +
<br>
 +
</div>
 +
</td>
 +
</tr>
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<tr>
 +
<td ColSpan="2" width="965">
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<div id="NAKAMI">
 +
<br>
 +
<div align="center">
 +
<strong>Our Sponsors</strong>
 +
<br><br>
 +
<a href="http://www.kit.ac.jp/"><img src="https://static.igem.org/mediawiki/2012/e/ea/KITkyoto.png" width="100" height="100"></a>
 +
<br>
 +
<br>
</div>
</div>
</div>
</div>
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</td>
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</div>
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</tr>
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</div>  
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</table>
</body>
</body>
</html>
</html>

Latest revision as of 12:49, 25 September 2012
TNFAIP3
PARTS

BP reaction by Invitrogen gateway system


1. PCR using primers containing the attB sequence.
2. Purify PCR product.
3. Add the following components to a 1.5 mL microcentrifuge tube at room temperature and mix:

attB PCR product75 ng/reaction (1-7 μL)
pDONR vector150ng/reaction (1 μL)
TE Bufferto 8 μL

4. Vortex BP ClonaseII enzyme mix briefly. Add 1 - 2 μL to the components above and mix well by vortexing and spin them down.
5. Incubate reaction at 25˚C for more than 1 hour.
6. Add 1 μL of 2 μg/μL Proteinase K solution and vortex briefly.
7. Incubate at 37˚C for 10 minutes.

LR reaction by Invitrogen gateway system


1. Add the following components to a 1.5 mL microcentrifuge tube at room temperature and mix:

Entry clones50-150 ng/reaction (1-7 μL)
Destination vector150ng/reaction (1 μL)
TE Bufferto 8 − 9 μL

2. Vortex LR ClonaseII enzyme mix briefly. Add 1 – 2 μL, to the components above and mix well by vortexing and spin down.
3. Incubate reaction at 25˚C for 16 hours.
4. Add 1 μL of 2 μg/μL Proteinase K solution and vortex briefly.

5. Incubate at 37˚C for 10 minutes.


Our Sponsors



Retrieved from "http://2012.igem.org/Team:KIT-Kyoto/kazukokekokko"