Team:KIT-Kyoto/kazukokekokko

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{{KIT-Kyoto.Header}}
 
{{KIT-Kyoto}}
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{{KIT-Kyoto.Header}}
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<!--段組み--!>
 
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<div id="NAKAMI">
 
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<div id="HIDARI">
 
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KIT-Kyoto2012
 
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<br>
 
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TWITTER
 
-
<br>
 
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Counter
 
<br>
<br>
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などを配置する。
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<div>
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    <ul class="navi">
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        <li>
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            <div class="category"><img src="https://static.igem.org/mediawiki/2012/1/1f/Side_labnotekit.jpg" width="150" height="30"></div>
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                <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-week5"><img src="https://static.igem.org/mediawiki/2012/e/e7/Side_week5kit.jpg" width="150" height="30"></a></li>
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                <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-week6"><img src="https://static.igem.org/mediawiki/2012/e/e8/Side_week6kit.jpg" width="150" height="30"></a></li>
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                <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-week7"><img src="https://static.igem.org/mediawiki/2012/b/b2/Side_weekkit7.jpg" width="150" height="30"></a></li>
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            <div class="category"><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Protocol"><img src="https://static.igem.org/mediawiki/2012/e/ee/Side_protocolkit.jpg" width="150" height="30"></a></div>
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                <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Meeting-june"><img src="https://static.igem.org/mediawiki/2012/9/92/Side_junekit.jpg" width="150" height="30"></a></li>
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                <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Meeting-july"><img src="https://static.igem.org/mediawiki/2012/f/f2/Side_julykit.jpg" width="150" height="30"></a></li>
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                <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Meeting-august"><img src="https://static.igem.org/mediawiki/2012/f/fa/Side_augkit.jpg" width="150" height="30"></a></li><li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Meeting-september"><img src="https://static.igem.org/mediawiki/2012/8/8b/Side_sepkit.jpg" width="150" height="30"></a></li>
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<br>
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</td>
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<td width="800px" height="510px" valign="top">
<div id="MIGI">
<div id="MIGI">
-
シンプルさ優先！アブスト、スポンサー、実験の概要の画像？？？？
+
<font color="#f5b1aa"><u>TNFAIP3</u></font>
<br>
<br>
-
↓Notebookのナビにはりつける
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<font color="#f5b1aa"><u>PARTS</u></font>
-
<ul class="acc">  
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-
  <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook"><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>
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-
      <li><div><a href="">Week9 ()</a></div></li>
+
-
      <li><div><a href="">Week10 ()</a></div></li>
+
-
    </ul>
+
-
  </li>
+
-
</ul>
+
-
<a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Protocol"><img src="" width="160" height="32" border="0" bordercolor="black">PROTOCOL</a>
+
-
<BR>
+
-
<ul class="acc1">
+
-
  <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Meeting"><img src="" width="160" height="32" class="menu_head" />MEETING</a>
+
-
    <ul class="fxmn">
+
-
    <li><div><a href="">First</a></div></li>
+
-
    <li><div><a href="">MAY</a></div></li>
+
-
    <li><div><a href="">June </a></div></li>
+
-
    <li><div><a href="">July </a></div></li>
+
-
    <li><div><a href="">August</a></div></li>
+
-
    <li><div><a href="">September</a></div></li>
+
-
    </ul>
+
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  </li>
+
-
</ul>
+
-
<a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Design"><img src="" width="160" height="32" border="0" bordercolor="black">DESIGN NOTE</a>
+
-
<BR>
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<br>
<br>
-
pfofile ex.<A href="https://2012.igem.org/Team:KIT-Kyoto/Team-Kazuko" >いちいち押すやつ</A>
+
<h2>BP reaction by Invitrogen gateway system</h2>
<br>
<br>
-
見出し <a href="http://midashi-maker.v-colors.com/">go</a>
+
1. PCR using primers containing the attB sequence.
<br>
<br>
-
<a href="http://www.colordic.org/w/"> 日本の伝統色</a>
+
2. Purify PCR product.
<br>
<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>
 +
<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>
 +
 +
<tr>
 +
<td ColSpan="2" width="965">
 +
<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>
 +
</td>
 +
</tr>
</div>  
</div>  
 +
</table>
 +
</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



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