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Team:KIT-Kyoto/kazukokekokko
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| + | {{KIT-Kyoto}} | ||
| + | {{KIT-Kyoto.Header}} | ||
<html> | <html> | ||
| - | < | + | <head> |
| - | < | + | <meta charset="UTF-8"> |
| - | + | <script type="text/javascript" src="http://ajax.googleapis.com/ajax/libs/jquery/1.4.2/jquery.min.js"></script> | |
| - | + | <script type="text/javascript"> | |
| - | < | + | $(function(){ |
| - | < | + | $("ul.menu").hide(); |
| - | + | $("div.category").click(function(){ | |
| - | + | $("ul.menu").slideUp(); | |
| + | if($("+ul",this).css("display")=="none"){ | ||
| + | $("+ul",this).slideDown(); | ||
| + | } | ||
| + | }); | ||
| + | }); | ||
| + | </script> | ||
| + | <style type="text/css"> | ||
| + | body { | ||
| + | font-family: Myriad, Helvetica, Arial, "Meiryo", "メイリオ", sans-serif; | ||
| + | _font-family: 'ＭＳ Ｐゴシック', sans-serif; | ||
| + | } | ||
| + | div.category { | ||
| + | margin:-top:5px; | ||
| + | height:40px; | ||
| + | line-height:40px; | ||
| + | cursor:pointer; | ||
| + | } | ||
| + | ul.navi{ | ||
| + | width: 150px; | ||
| + | margin: 0px; | ||
| + | } | ||
| + | ul.navi, ul.menu { | ||
| + | padding:0; | ||
| + | margin:0; | ||
| + | list-style: none; | ||
| + | } | ||
| + | ul.menu li{ | ||
| + | padding-left:15px; | ||
| + | margin:0; | ||
| + | list-style: none; | ||
| + | } | ||
| + | div.category { | ||
| + | margin-top: 5px; | ||
| + | padding-left: 15px; | ||
| + | height: 32px; | ||
| + | line-height: 40px; | ||
| + | } | ||
| + | ul.menu a { | ||
| + | display: block; | ||
| + | height: 32px; | ||
| + | line-height: 10px; | ||
| + | } | ||
| + | </style> | ||
| + | </head> | ||
| + | <body> | ||
| + | <table border="0" width="965px" align="center"><tr><td width="165px" valign="top" | ||
| + | |||
| + | align="left"><div id="HIDARI"> | ||
| + | |||
<br> | <br> | ||
| - | <img src="https://static.igem.org/mediawiki/2012/ | + | <div> |
| - | <a href="https://2012.igem.org/Team:KIT-Kyoto/ | + | <ul class="navi"> |
| - | <img src="https://static.igem.org/mediawiki/2012/ | + | <li> |
| - | <a href="https://2012.igem.org/Team:KIT-Kyoto/Team- | + | <div class="category"><img src="https://static.igem.org/mediawiki/2012/1/1f/Side_labnotekit.jpg" width="150" height="30"></div> |
| - | <img src="https://static.igem.org/mediawiki/2012/ | + | <ul class="menu"> |
| - | <a href="https://2012.igem.org/Team:KIT-Kyoto/ | + | <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> |
| + | <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-week2"><img src="https://static.igem.org/mediawiki/2012/9/94/Side_week2kit.jpg" width="150" height="30"></a></li> | ||
| + | <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-week3"><img src="https://static.igem.org/mediawiki/2012/e/e0/Side_week3kit.jpg" width="150" height="30"></a></li> | ||
| + | <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> | ||
| + | <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> | ||
| + | <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> | ||
| + | <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> | ||
| + | </ul> | ||
| + | </li> | ||
| + | <li> | ||
| + | <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> | ||
| + | </li> | ||
| + | <li> | ||
| + | <div class="category"><img src="https://static.igem.org/mediawiki/2012/6/61/Side_meetingkit.jpg" width="150" height="30"></div> | ||
| + | <ul class="menu"> | ||
| + | <li><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Meeting-may"><img src="https://static.igem.org/mediawiki/2012/5/5d/Side_maykit.jpg" width="150" height="30"></a></li> | ||
| + | <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> | ||
| + | <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> | ||
| + | <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> | ||
| + | </ul> | ||
| + | </li> | ||
| + | <li> | ||
| + | <div class="category"><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-Design"><img src="https://static.igem.org/mediawiki/2012/8/8f/Side_designnotekit.jpg" width="150" height="30"></a></div> | ||
| + | </li> | ||
| + | </ul> | ||
| + | </div> | ||
<br> | <br> | ||
| - | < | + | </td> |
| - | < | + | <td width="800px" height="510px" valign="top"> |
| - | + | <div id="MIGI"> | |
| - | < | + | <font color="#f5b1aa"><u>TNFAIP3</u></font> |
| - | < | + | |
| - | < | + | |
<br> | <br> | ||
| - | < | + | <font color="#f5b1aa"><u>PARTS</u></font> |
| - | < | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
<br> | <br> | ||
| - | < | + | <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> | ||
| + | <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> | ||
| - | |||
| - | |||
| - | |||
| - | |||
<br> | <br> | ||
| - | + | </div> | |
| + | </td> | ||
| + | </tr> | ||
| + | |||
| + | <tr> | ||
| + | <td ColSpan="2" width="965"> | ||
| + | <div id="NAKAMI"> | ||
<br> | <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> | ||
| - | |||
<br> | <br> | ||
| - | + | ||
| + | </div> | ||
| + | </div> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </div> | ||
| + | </table> | ||
</body> | </body> | ||
</html> | </html> | ||
Latest revision as of 12:49, 25 September 2012
|
TNFAIP3
PARTS BP reaction by Invitrogen gateway system1. 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:
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 system1. Add the following components to a 1.5 mL microcentrifuge tube at room temperature and mix:
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. |
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