Team:KIT-Kyoto/kazukokekokko

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align="left"><div id="HIDARI">
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KIT-Kyoto2012
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<br>
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TWITTER
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                <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-week1"><img src="https://static.igem.org/mediawiki/2012/1/1b/Side_week1kit.jpg" width="150" height="30"></a></li>
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                <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-week4"><img src="https://static.igem.org/mediawiki/2012/3/31/Side_week4kit.jpg" width="150" height="30"></a></li>
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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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シンプルさ優先!アブスト、スポンサー、実験の概要の画像????
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<div id="MIGI">
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<font color="#f5b1aa"><u>TNFAIP3</u></font>
<br>
<br>
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↓Notebookのナビにはりつける
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<font color="#f5b1aa"><u>PARTS</u></font>
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<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>
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      <li><div><a href="">Week10 ()</a></div></li>
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    </ul>
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<a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Protocol"><img src="" width="160" height="32" border="0" bordercolor="black">PROTOCOL</a>
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<BR>
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<ul class="acc1">
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  <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Meeting"><img src="" width="160" height="32" class="menu_head" />MEETING</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="">MAY</a></div></li>
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    <li><div><a href="">June </a></div></li>
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    <li><div><a href="">July </a></div></li>
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    <li><div><a href="">August</a></div></li>
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    <li><div><a href="">September</a></div></li>
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    </ul>
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  </li>
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</ul>
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<a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Design"><img src="" width="160" height="32" border="0" bordercolor="black">DESIGN NOTE</a>
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<BR>
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<br>
<br>
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<h2>BP reaction by Invitrogen gateway system</h2>
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<strong>Q.1 Would any of your project ideas raise safety issues in terms of:
+
<br>
<br>
-
1.researcher safety,
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1. PCR using primers containing the attB sequence.
<br>
<br>
-
2.public safety, or
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2. Purify PCR product.
<br>
<br>
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3.environmental safety?</strong>
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3. Add the following components to a 1.5 mL microcentrifuge tube at room temperature and mix:
<br><br>
<br><br>
-
No issues of researcher safety, public safety or environmental safety raised up during our KIT-Kyoto iGEM 2012 project. We used nucleic acid donor or host, which follow the guidelines of the safety standards required by biosafety level 1.
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<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>
<br>
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In our project, we plan to establish transgenic flies. They will be produced and kept in P1 room and fly room in which fly trap and double-entry doors are equipped. By these safety equipments, even if they escape from the culture bottle to the room, we can block them flying out of the room.
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4. Vortex BP ClonaseII enzyme mix briefly. Add 1 - 2 μL to the components above and mix well by vortexing and spin them down.
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<br><br>
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<strong>Q.2 Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes,
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<br>
<br>
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1.did you document these issues in the Registry?
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5. Incubate reaction at 25˚C for more than 1 hour.
<br>
<br>
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2.How did you manage to handle the safety issue?
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6. Add 1 μL of 2 μg/μL Proteinase K solution and vortex briefly.
<br>
<br>
-
3.How could other teams learn from your experience?</strong>
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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>
<br><br>
-
 
+
<Table Border Cellspacing="0">
-
<strong>Q.3 Is there a local biosafety group, committee, or review board at your institution?
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<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>
<br>
-
1.If yes, what does your local biosafety group think about your project?
+
5. Incubate at 37˚C for 10 minutes.
<br>
<br>
-
2.If no, which specific biosafety rules or guidelines do you have to consider in your country?</strong>
+
<br>
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<br><br>
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</div>
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A. Yes. We have bio safety group at KIT, and our project has been monitored by them. The leader of this biological safety group was Prof. Yamaguchi. Furthermore, there is an upper level committee of the recombinant DNA experiments at Kyoto Institute of Technology. Our project has also received permission from this committee. All material handled or distributed are non-hazardous and non-infectious. It meets with all safety standards requested by biosafety level 1. In addition, Prof. Yamaguchi and other supervisors and instructors fully support the work done by this iGEM team.
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</td>
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<br><br>
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</tr>
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<strong>Q.4 Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?</strong>
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<tr>
 +
<td ColSpan="2" width="965">
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<div id="NAKAMI">
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<br>
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<div align="center">
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<strong>Our Sponsors</strong>
<br><br>
<br><br>
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A. We have two suggestions.
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<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>
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Firstly, to understand how ethical a project is and how safe equipments are, we should make Bio-safety Quiz, and the iGEM teams cannot get description kit until they complete the quiz.
 
<br>
<br>
-
Secondly, we should supply check list template of Bio-safety in wiki, and each iGEM team should fill out the check list on their experiment notebooks as a template and check it at the beginning and ending of the diary experiments.
 
-
<br><br><br>
 
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<strong>Lecture class about experiments of genetic engineering</strong>
 
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<br><br>
 
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We took a lecture class about genetic engineering because we conduct experiments involving recombinant DNA technic for our project of iGEM. We learned its guidelines and laws
 
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received an instruction.
 
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<br>
 
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<img src="https://static.igem.org/mediawiki/2012/e/e8/P1040319.JPG" width="300" height="200" align="right">
 
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</td></tr>
 
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</body>
</body>
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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