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| - | >[[Team:Chiba/jk/β|送受信班]]
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| | | | |
| - | ==Week 2==
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| - | ===24 August, 2008===
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| - | :(22/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| |
| - | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
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| - | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
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| - | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
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| - | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
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| - |
| |
| - |
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| - | :--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
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| - | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
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| - | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
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| - | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
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| - | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
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| - |
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| - |
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| - | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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| - | <tr>
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| - | <td width="257">Sample No</td>
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| - | <td>5</td><td>6</td><td>7</td><td>8</td>
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| - | </tr>
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| - | <tr>
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| - | <td>BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)</td>
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| - | <td>1</td><td>-</td><td>-</td><td>-</td>
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| - | </tr>
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| - | <tr>
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| - | <td>BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)</td>
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| - | <td>-</td><td>1</td><td>-</td><td>-</td>
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| - | </tr><tr>
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| - | <td>BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)</td>
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| - | <td>-</td><td>-</td><td>1</td><td>-</td>
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| - | </tr><tr>
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| - | <td>BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)</td>
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| - | <td>-</td><td>-</td><td>-</td><td>1</td>
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| - | </tr>
| |
| - | <tr>
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| - | <td>EcoRⅠ</td>
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| - | <td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Buffer 4</td>
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| - | <td>0.9</td><td>0.9</td><td>0.9</td><td>0.9</td>
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| - | </tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>8</td><td>8</td><td>8</td><td>8</td>
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| - | </tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>10</td><td>10</td><td>10</td><td>10</td>
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| - | </tr>
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| - | </table>
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| - |
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| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
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| - | {|align="justify"
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| - | |[[Image:Chiba-0824-1.JPG]]
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| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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| - | <tr>
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| - | <td width="257">Sample No</td>
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| - | <td>1~8</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Sample DNA</td>
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| - | <td>1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Loading Dye</td>
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| - | <td>1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>4</td>
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| - | </tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>6</td>
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| - | </tr>
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| - | </table>
| |
| - | :From left;
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| - | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> ?
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| - | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006) -> OK
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| - | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) -> OK
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| - | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006) -> OK
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| - | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> OK
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| - | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006) -> OK
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| - | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) -> OK
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| - | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006) -> None?
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| - | |}
| |
| - | {|align="justify"
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| - | |[[Image:Chiba-0824-2.JPG]]
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| - | |
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| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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| - | <tr>
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| - | <td width="257">Sample No</td>
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| - | <td>5~8</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Sample DNA</td>
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| - | <td>9</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Loading Dye</td>
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| - | <td>2</td></tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>12</td>
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| - | </tr>
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| - | </table>
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| - |
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| - | From the left;
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| - | （single digestuion）
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| - | ::5, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> OK
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| - | ::6, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006) -> OK
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| - | ::7, BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) -> 3400bp:Bad, 2200bp:OK
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| - | ::8, BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006) -> OK
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| - | |}
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| - |
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| - | ===25 August, 2008===
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| - | :(24/8)--->'''[[Team:Chiba/protocol/digestion|Digestion]]'''
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| - | ::#Double Digestion:BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
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| - | ::#Double Digesiton:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
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| - | ::#Single Digestion:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
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| - | ::#BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
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| - |
| |
| - | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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| - | <tr>
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| - | <td width="257">Sample No</td>
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| - | <td>1</td><td>2</td><td>3</td><td>4</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Sample DNA</td>
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| - | <td>5</td><td>5</td><td>1</td><td>1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>EcoRⅠ</td>
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| - | <td>0.5</td><td>o.5</td><td>-</td><td>-</td>
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| - | </tr>
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| - | <tr>
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| - | <td>SpeⅠ</td>
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| - | <td>0.5</td><td>0.5</td><td>-</td><td>-</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Buffer 1</td>
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| - | <td>1</td><td>1</td><td>-</td><td>-</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Buffer 3</td>
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| - | <td>-</td><td>-</td><td>1</td><td>-</td>
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| - | </tr>
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| - | <tr>
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| - | <td>BSA</td>
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| - | <td>1</td><td>1</td><td>-</td><td>-</td>
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| - | </tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>2</td><td>2</td><td>7.5</td><td>-</td>
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| - | </tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>10</td><td>10</td><td>10</td><td>1</td>
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| - | </tr>
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| - | </table>
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| - |
| |
| - | :--->[[Team:Chiba/protocol/gelcheck|Gel Check]]
| |
| - | {|align="justify"
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| - | |[[Image:Chiba-0825-1.JPG]]
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| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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| - | <tr>
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| - | <td width="257">Sample No</td>
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| - | <td>1~3</td><td>4</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Sample DNA</td>
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| - | <td>10</td><td>1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Loading Dye</td>
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| - | <td>2</td><td>1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>-</td><td>4</td>
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| - | </tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>12</td><td>6</td>
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| - | </tr>
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| - | </table>
| |
| - | :From right;
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| - | ::#Double Digestion:BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> OK
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| - | ::#Double Digesiton:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> ?(low)
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| - | ::#Single Digestion:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> ?(low)
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| - | ::#BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> OK(low)
| |
| - | |}
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| - |
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| - |
| |
| - | --->[[Team:Chiba/protocol/digestion|Digestion]]
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| - | (Double Digestion)
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| - | ::#BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
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| - | ::#BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
| |
| - | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
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| - |
| |
| - | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
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| - | <td width="257">Sample No</td>
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| - | <td>1</td><td>2</td><td>3</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Sample DNA</td>
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| - | <td>33ng/μl×60μl</td><td>33ng/μl×60μl</td><td>100ng/μl×20μl</td>
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| - | </tr>
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| - | <tr>
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| - | <td>PstⅠ</td>
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| - | <td>2</td><td>2</td><td>2</td>
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| - | </tr>
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| - | <tr>
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| - | <td>XbaⅠ</td>
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| - | <td>2</td><td>2</td><td>-</td>
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| - | </tr>]
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| - | <td>SpeⅠ</td>
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| - | <td>-</td><td>-</td><td>2</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Buffer 2</td>
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| - | <td>-</td><td>-</td><td>5</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Buffer 3</td>
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| - | <td>10</td><td>10</td><td>-</td>
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| - | </tr>
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| - | <tr>
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| - | <td>BSA</td>
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| - | <td>10</td><td>10</td><td>5</td>
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| - | </tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>16</td><td>16</td><td>16</td>
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| - | </tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>100</td><td>100</td><td>52</td>
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| - | </tr>
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| - | </table>
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| - |
| |
| - | ===26 August, 2008===
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| - | :(8/25)--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
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| - | {|align="justify"
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| - | |[[Image:Chiba-0825-2.JPG]]
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| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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| - | <tr>
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| - | <td width="257">Sample No</td>
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| - | <td>1~3</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Sample DNA</td>
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| - | <td>30</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Loading Dye</td>
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| - | <td>6</td>
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| - | </tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>36</td>
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| - | </tr>
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| - | </table>
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| - | ::1, BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
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| - | :::Insert:609bp -> OK
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| - | |}
| |
| - | {|align="justify"
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| - | |[[Image:Chiba-0825-3.JPG]]
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| - | |
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| - | ::2, BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
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| - | :::Insert:609bp -> OK
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| - | |}
| |
| - | {|align="justify"
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| - | |[[Image:Chiba-0825-4.JPG]]
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| - | |
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| - | ::3, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
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| - | ::: -> OK
| |
| - | |}
| |
| - |
| |
| - |
| |
| - | --->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
| |
| - | :::#5μl
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| - | :::#5μl
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| - | :::#27μl
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| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0825-5.JPG]]
| |
| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
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| - | <td width="257">Sample No</td>
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| - | <td>1~3</td>
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| - | </tr>
| |
| - | <tr>
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| - | <td>Sample DNA</td>
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| - | <td>2</td>
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| - | </tr>
| |
| - | <tr>
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| - | <td>Loading Dye</td>
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| - | <td>2</td></tr>
| |
| - | <tr>
| |
| - | <td>H2O</td>
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| - | <td>8</td></tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | :From left;
| |
| - | ::#insert:C0178 -> OK
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| - | ::#insert:C0170 -> OK?
| |
| - | ::#vector:J04500 -> OK
| |
| - | |}
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/ligation/dephosphorylation|SAP]]'''
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
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| - | <td width="257">Sample No</td>
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| - | <td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>26</td>
| |
| - | </tr>
| |
| - | <tr>
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| - | <td>SAP</td>
| |
| - | <td>1</td></tr>
| |
| - | <tr>
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| - | <td>SAP Buffer</td>
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| - | <td>3</td></tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>30μL</td></tr>
| |
| - | </table>
| |
| - |
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
| |
| - | :::Sample No.3->9μl
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0826.JPG]]
| |
| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No.3</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6</td></tr>
| |
| - | </table>
| |
| - | ::Sample No.3 -> OK ･･･30ng?
| |
| - | |}
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/ligation/ligation|Ligation]]'''
| |
| - | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>(1)</td><td>(2)</td><td>(3)</td><td>(4)</td><td>(5)</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>insert①C0178</td>
| |
| - | <td>1</td><td>-</td><td>-</td><td>1</td><td>-</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>insert②C0170</td>
| |
| - | <td>-</td><td>1</td><td>-</td><td>-</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>vector③J04500</td>
| |
| - | <td>-</td><td>-</td><td>2</td><td>2</td><td>2</td>
| |
| - | </tr>]
| |
| - | <td>ligase</td>
| |
| - | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer</td>
| |
| - | <td>2</td><td>2</td><td>2</td><td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>6</td><td>6</td><td>5</td><td>4</td><td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| |
| - | ::#insert①C0178
| |
| - | ::#insert②C0170
| |
| - | ::#vector③J04500
| |
| - | ::#insert①C0178+vector③J04500
| |
| - | ::#insert②C0170+vector③J04500
| |
| - |
| |
| - |
| |
| - |
| |
| - | ===27 August, 2008===
| |
| - | [[Team:Chiba/protocol/digestion|Digestion Test]]
| |
| - | ::Sample DNA : BBa_R0010[http://partsregistry.org/Part:BBa_R0010]
| |
| - |
| |
| - | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>1</td><td>2</td><td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>PstⅠ</td>
| |
| - | <td>-</td><td>-</td><td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ</td>
| |
| - | <td>-</td><td>0.5</td><td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer 3</td>
| |
| - | <td>-</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>-</td><td>-</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>-</td><td>7.5</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>1</td><td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0827.JPG]]
| |
| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>1</td><td>2</td><td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>1</td><td>5</td><td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>H2O</td>
| |
| - | <td>4</td><td>-</td><td>-</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6</td><td>6</td><td>6</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | :From left;<br>
| |
| - | :#BBa_R0010
| |
| - | :#Single Digestion
| |
| - | :#Double Digestion
| |
| - | |}
| |
| - |
| |
| - |
| |
| - |
| |
| - | :(26/8)--->'''[[Team:Chiba/protocol/PCR|Colony-PCR]]'''
| |
| - | ::Colony PCR of 5 colonies from ligation plates (26/8-(4),(5)) and one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
| |
| - |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">DNA Template</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dNTP mix</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Foward Primer</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Reverse Primer</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>DNA polymerase</td>
| |
| - | <td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Thermopol Buffer</td>
| |
| - | <td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>28.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>50μL</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - |
| |
| - | :95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃
| |
| - |
| |
| - |
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0827-2.JPG]]
| |
| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample DNA</td>
| |
| - | <td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td></tr>
| |
| - | </table>
| |
| - | :From left;<br>
| |
| - | :1~5 insert:C0170 + vector:J04500 -> OK
| |
| - | :6 F2620･･Positive Control -> OK
| |
| - | :7~11 insert:C0178 + vector:J04500 -> OK
| |
| - | |}
| |
| - |
| |
| - |
| |
| - | :--->(28/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Miniprep]]'''
| |
| - |
| |
| - |
| |
| - |
| |
| - | '''[[Team:Chiba/protocol/transformation|Transformation]]'''
| |
| - | :competent cells : XL10G
| |
| - | ::*BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
| |
| - | ::*BBa_C0062(2006) [http://partsregistry.org/Part:BBa_C0062] -> no colony on plates
| |
| - | ::*BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
| |
| - | ::*BBa_C0179(2006) [http://partsregistry.org/Part:BBa_C0179]
| |
| - | ::*BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
| |
| - | ::*BBa_S03156(2006) [http://partsregistry.org/Part:BBa_S03156]
| |
| - | ::*BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
| |
| - | ::*BBa_S03158(2006) [http://partsregistry.org/Part:BBa_S03158]
| |
| - | ::*BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
| |
| - | ::*BBa_S0316(2006) [http://partsregistry.org/Part:BBa_S03160]
| |
| - |
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| |
| - |
| |
| - |
| |
| - | :--->(29/8)'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - |
| |
| - |
| |
| - |
| |
| - | ===28 August, 2008===
| |
| - | :(27/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]
| |
| - | ::#insert:C0170 + vector:J04500
| |
| - | ::#insert:C0178 + vector:J04500
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/digestion|Digestion Test]]'''
| |
| - | ::#insert:C0170 + vector:J04500
| |
| - | ::#insert:C0178 + vector:J04500
| |
| - |
| |
| - | <table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>3</td><td>4</td><td>5</td><td>6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>insert:C0170 + vector:J04500 </td>
| |
| - | <td>1</td><td>-</td><td>1</td><td>-</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>insert:C0178 + vector:J04500 </td>
| |
| - | <td>-</td><td>1</td><td>-</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ</td>
| |
| - | <td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>SpeⅠ</td>
| |
| - | <td>-</td><td>-</td><td>0.1</td><td>0.1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer 3</td>
| |
| - | <td>1</td><td>1</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>-</td><td>-</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>7.8</td><td>7.8</td><td>6.8</td><td>6.8</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>10</td><td>10</td><td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - |
| |
| - |
| |
| - | :--->(29/8)'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0828.JPG]]
| |
| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>1,2</td><td>3~6</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>1</td><td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>1</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>H2O</td>
| |
| - | <td>4</td><td>-</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6</td><td>12</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | :From left;
| |
| - | ::# -> OK
| |
| - | ::# -> OK
| |
| - | ::# -> OK
| |
| - | ::# -> OK
| |
| - | ::# -> OK
| |
| - | ::# -> OK
| |
| - | |}
| |
| - |
| |
| - | ===29 August, 2008===
| |
| - | :--->'''[[Team:Chiba/protocol/digestion|Digestion Test]]'''
| |
| - | ::#BBa_R0010(2007) [http://partsregistry.org/Part:BBa_R0010]
| |
| - | ::#BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
| |
| - | ::#BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
| |
| - | ::#BBa_C0179(2006) [http://partsregistry.org/Part:BBa_C0179]
| |
| - | ::#BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
| |
| - | ::#BBa_S03156(2006) [http://partsregistry.org/Part:BBa_S03156]
| |
| - | ::#BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
| |
| - | ::#BBa_S03158(2006) [http://partsregistry.org/Part:BBa_S03158]
| |
| - | ::#BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
| |
| - | ::#BBa_S03160(2006) [http://partsregistry.org/Part:BBa_S03160]
| |
| - |
| |
| - |
| |
| - | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>S1~10</td><td>W1</td><td>W2~10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>1</td><td>4</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>PstⅠ</td>
| |
| - | <td>-</td><td>0.075</td><td>0.1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>EcoRⅠ</td>
| |
| - | <td>0.1</td><td>0.075</td><td>0.1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer 3</td>
| |
| - | <td>1</td><td>0.75</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>-</td><td>0.75</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>7.9</td><td>4.35</td><td>5.8</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>10</td><td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0829.JPG]]
| |
| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No.</td>
| |
| - | <td>1~10</td><td>S1~S10,W1~W10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>1</td><td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>1</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6</td><td>12</td></tr>
| |
| - | </table>
| |
| - | :From left;<br>
| |
| - | ::1~5,S1,S5,W1~W5
| |
| - | |}
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0829-2.JPG]]
| |
| - | |
| |
| - | :From left;<br>
| |
| - | ::6~10,S6~S10,W6~W10
| |
| - | |}
| |
| - |
| |
| - |
| |
| - |
| |
| - | :(27/8)--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0829-3.JPG]]
| |
| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample DNA</td>
| |
| - | <td>0.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>4.5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6μL</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | ::R0010 -> OK
| |
| - | |}
| |
| - |
| |
| - |
| |
| - | '''[[Team:Chiba/protocol/digestion|Digestion]]'''
| |
| - | ::#BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
| |
| - | ::#BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
| |
| - | ::#BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
| |
| - | ::#BBa_R0010(2006) [http://partsregistry.org/Part:BBa_R0010]
| |
| - |
| |
| - |
| |
| - | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>1~3</td><td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>100ng/μL×20</td><td>200ng/μL×30</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>PstⅠ</td>
| |
| - | <td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>XbaⅠ</td>
| |
| - | <td>2</td><td>-</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>SpeⅠ</td>
| |
| - | <td>-</td><td>2</td></tr>
| |
| - | <tr>
| |
| - | <td>Buffer 2</td>
| |
| - | <td>-</td><td>5</td></tr>
| |
| - | <tr>
| |
| - | <tr>
| |
| - | <td>Buffer 3</td>
| |
| - | <td>5</td><td>-</td></tr>
| |
| - | <tr>
| |
| - | <td>BSA</td>
| |
| - | <td>5</td><td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>16</td><td>6</td></tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>50</td><td>50</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0829-4.JPG]]
| |
| - | |
| |
| - | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No.</td>
| |
| - | <td>1~3</td><td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>10</td><td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>-</td><td>7</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td><td>12</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | :insert-I9026 : 685bp -> None<br>
| |
| - | |}
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0829-5.JPG]]
| |
| - | |
| |
| - | :insert-I9030 : 745bp -> None<br>
| |
| - | |}
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0829-6.JPG]]
| |
| - | |
| |
| - | :insert-S03154 : 685bp -> None<br>
| |
| - | |}
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0829-7.JPG]]
| |
| - | |
| |
| - | :vector-R0010 : 2079bp -> OK<br>
| |
| - | |}
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
| |
| - | :::->15μL
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/ligation/dephosphorylation|SAP]]'''
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>4</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>15</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>SAP</td>
| |
| - | <td>2</td></tr>
| |
| - | <tr>
| |
| - | <td>SAP Buffer</td>
| |
| - | <td>3</td></tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>10</td></tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>30μL</td></tr>
| |
| - | </table>
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
| |
| - |
| |
| - | ::--->Suspension! (We got information, by the team-output, to the effect that the density of the sample is too low.)
| |
| - |
| |
| - |
| |
| - | ===30 August, 2008===
| |
| - | :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| |
| - | :competent cells : XL10G
| |
| - | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
| |
| - | ::*BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
| |
| - | ::*BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
| |
| - | ::*BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
| |
| - | ::*BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
| |
| - | ::*BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
| |
| - | ::*BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
| |
| - | ::*BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
| |
| - |
| |
| - | :--->(31/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| |
| - |
| |
| - |
| |
| - |
| |
| - | :(29/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| |
| - | ::#insert:C0170 + vector:J04500
| |
| - | ::#insert:C0178 + vector:J04500
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/digestion|Digestion Test]]'''
| |
| - | ::*insert:C0170 + vector:J04500 -> Sample No.1~4
| |
| - | ::*insert:C0178 + vector:J04500 -> Sample No.5~8
| |
| - | ::*BBa_T9002(2007)[http://partsregistry.org/Part:BBa_T9002] -> Sample No.9
| |
| - | ::*Single Digestion of Sample No.1~9 -> Sample No.10~18
| |
| - | ::*Double Digestion of Sample No.1~9 -> Sample No.19~27
| |
| - |
| |
| - | :<table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="270">Sample No.</td><td>Single : 10~18</td><td>Double : 19~27</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td><td>1</td><td>5</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>XbaⅠ</td><td>0.1</td><td>0.1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>SpeⅠ</td><td>-</td><td>0.1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Buffer 2</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>BSA</td><td>1</td><td>1</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td><td>6.9</td><td>2.8</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td><td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - |
| |
| - |
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0830.JPG]]
| |
| - | |
| |
| - | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| - | <tr>
| |
| - | <td width="257">Sample No</td>
| |
| - | <td>1~9</td><td>10~18</td><td>19~27</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>1</td><td>10</td><td>10</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>1</td><td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>4</td><td>-</td><td>-</td></tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>6</td><td>12</td><td>12</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | :From left;<br>
| |
| - | ::Sanple No.1~13
| |
| - | ::-> OK
| |
| - | |}
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0830-2.JPG]]
| |
| - | |
| |
| - | :From left;<br>
| |
| - | ::Sample No.14~17,24~16
| |
| - | ::14 -> OK
| |
| - | ::15,16 -> Bad
| |
| - | ::17 -> None
| |
| - | ::24~16 -> Bad
| |
| - | |}
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0830-3.JPG]]
| |
| - | |
| |
| - | :From left;<br>
| |
| - | ::Sample No.18,27,19~23
| |
| - | ::18,27 -> Bad
| |
| - | ::19~22 -> OK
| |
| - | ::23 -> OK??
| |
| - | |}
| |
| - |
| |
| - | ===[[Team:Chiba/jk/β/weak 3|>next week]]===
| |
| - |
| |
| - | {| style="color:white;background-color:Maroon" cellpadding="3" cellspacing="3" border="1" bordercolor="white" width="100%" align="center"
| |
| - | !align="center"|[[Team:Chiba/j|ホーム]]
| |
| - | !align="center"|[[Team:Chiba/Team/j|メンバー紹介]]
| |
| - | !align="center"|[[Team:Chiba/Project/j|プロジェクト紹介]]
| |
| - | !align="center"|[[Team:Chiba/Parts/j|Parts Submitted to the Registry]]
| |
| - | !align="center"|[[Team:Chiba/Modeling/j|モデリング]]
| |
| - | !align="center"|[[Team:Chiba/Internal|ノート]]
| |
| - | |}
| |
"
