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| - | >[[Team:Chiba/jk/β|送受信班]]
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| | | | |
| - | ==Week 3==
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| - | ===31 August, 2008===
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| - | :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
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| - | :Competent Cells : XL10G
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| - | ::*Plac+RBS+LasI
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| - | ::*Plac+RBS+RhlI
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| - |
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| - | '''[[Team:Chiba/protocol/digestion|Digestion]]'''
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| - | ::#BBa_I9026[http://partsregistry.org/Part:BBa_I9026](2007)
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| - | ::#BBa_I9030[http://partsregistry.org/Part:BBa_I9030](2006)
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| - | ::#BBa_S03154[http://partsregistry.org/Part:BBa_S03154](2007)
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| - | ::#BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
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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>1~3</td><td>4</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Sample DNA</td><td>12</td><td>10</td>
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| - | </tr>
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| - | <tr>
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| - | <td>PstⅠ</td>
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| - | <td>0.2</td><td>0.2</td>
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| - | </tr>
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| - | <tr>
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| - | <td>XbaⅠ</td><td>0.2</td><td>-</td>
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| - | </tr>
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| - | <tr>
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| - | <td>SpeⅠ</td><td>-</td><td>0.4</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>1.5</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Buffer 3</td>
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| - | <td>2</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>2</td><td>1.5</td>
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| - | </tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>3.6</td><td>1.4</td></tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>20</td><td>15</td>
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| - | </tr>
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| - | </table>
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| - |
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| - |
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| - | :--->(1/9)'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
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| - |
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| - |
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| - | :(30/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
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| - |
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| - |
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| - | :--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
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| - | :*Plac+RBS+RhlI Sample No.2~5
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| - | :*BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
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| - |
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| - | <table width="600" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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| - | <tr>
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| - | <td width="257">Sample</td>
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| - | <td>Single Digesiton 2~5</td><td>Double Digestion 2~5</td><td>Single Digestion T9002①</td><td>Single Digesiton T9002②</td><td><Double Digestion T9002①②</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Sample DNA</td><td>1</td><td>3</td><td>3</td><td>1</td><td>3</td>
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| - | </tr>
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| - | <tr>
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| - | <td>XbaⅠ</td><td>0.1</td><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>SpeⅠ</td><td>-</td><td>0.1</td><td>-</td><td>-</td><td>0.1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Buffer 2</td>
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| - | <td>0.9</td><td>0.8</td><td>0.9</td><td>0.9</td><td>0.8</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>1</td><td>1</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>7</td><td>5</td><td>5</td><td>7</td><td>5</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><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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| - |
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| - | {|align="justify"
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| - | |[[Image:Chiba-0831-p87.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>Plac+RBS+RhlI Sample No.2~5, BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①②
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| - | </td><td>Single & Double Digestion</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><td>10</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><td>2</td></tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>4</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>6</td><td>12</td>
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| - | </tr>
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| - | </table>
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| - | :From Left
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| - | ::*Single Digestion Plac+RBS+RhlI 2~5
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| - | ::*Single Digestion BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
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| - | ::*Double Digestion Plac+RBS+RhlI 2~5
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| - | ::*Double Digestion BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
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| - | ::*BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
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| - | |}
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| - |
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| - | ===1 September, 2008===
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| - | :(31/8)--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
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| - | {|align="justify"
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| - | |[[Image:Chiba-0901.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>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>20</td><td>15</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Loading Dye</td>
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| - | <td>4</td><td>3</td></tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>24</td><td>18</td>
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| - | </tr>
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| - | </table>
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| - | :From left;
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| - | ::insert-1(I9026)
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| - | ::insert-2(I9030)
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| - | |}
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| - | {|align="justify"
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| - | |[[Image:Chiba-0901-2.JPG]]
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| - | |
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| - | ::insert-3(S03154)
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| - | |}
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| - | {|align="justify"
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| - | |[[Image:Chiba-0901-3.JPG]]
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| - | |
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| - | ::vector-4(R0010)
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| - | |}
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| - |
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| - | :--->zymo
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| - | ::vector-4(R0010)
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| - | ::-> 15μL
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| - |
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| - | :--->SAP
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| - | ::vectr-4(R0010)
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| - |
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| - | :--->Zymo
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| - | ::1 -> 7μL
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| - | ::2 -> 7
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| - | ::3 -> 7
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| - | ::4 -> 20
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| - |
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| - |
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| - |
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| - | '''[[Team:Chiba/protocol/transformation|Transformation]]'''
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| - | :Competent cells : BW ΔFliC 40μL
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| - | ::*Plac+RBS+LasI : Sample No.L1~L4
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| - | ::*Plac+RBS+RhlI : Sample No.R1~R4
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| - |
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| - | :--->(2/9)OD
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| - |
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| - | ===2 September, 2008===
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| - | :(31/8)--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
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| - |
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| - | {|align="justify"
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| - | |[[Image:Chiba-0902.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>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>3</td><td>3</td><td>3</td><td>3</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>2</td><td>2</td><td>2</td>
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| - | </tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>7</td><td>7</td><td>7</td><td>7</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>12</td><td>12</td><td>12</td>
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| - | </tr>
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| - | </table>
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| - | :From left;
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| - | ::I9026
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| - | ::I9030
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| - | ::S03154
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| - | |}
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| - |
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| - |
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| - |
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| - | :(1/9)--->'''[[Team:Chiba/protocol/PCR|Colony-PCR]]'''
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| - | ::Colony PCR of 8 colonies from ligation plates (1/9-(1),(2)) and one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
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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">DNA Template</td>
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| - | <td>1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>dNTP mix</td>
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| - | <td>5</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Foward Primer</td>
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| - | <td>0.3</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Reverse Primer</td>
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| - | <td>0.3</td>
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| - | </tr>
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| - | <tr>
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| - | <td>DNA polymerase TAQ</td>
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| - | <td>0.5</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Thermopol Buffer</td>
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| - | <td>3</td>
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| - | </tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>20.5</td>
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| - | </tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>30μL</td>
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| - | </tr>
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| - | </table>
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| - |
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| - |
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| - | :95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃
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| - |
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| - |
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| - | --->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
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| - |
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| - | {|align="justify"
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| - | |[[Image:Chiba-0902-2.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 DNA</td>
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| - | <td>1</td></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>
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| - | :From left;
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| - | :*Plac+RBS+RhlI+LVA
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| - | ::R1 -> OK
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| - | ::R2 -> Bad
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| - | ::R3~R7 -> OK
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| - | ::R8 -> Bad
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| - | |}
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| - |
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| - | {|align="justify"
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| - | |[[Image:Chiba-0902-3.JPG]]
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| - | |
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| - | :From left;
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| - | :*Plac+RBS+CinI+LVA
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| - | ::C1,C2 -> OK
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| - | ::C3 -> Bad
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| - | ::C4~C6 -> OK
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| - | |}
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| - |
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| - | {|align="justify"
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| - | |[[Image:Chiba-0902-4.JPG]]
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| - | |
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| - | :From left;
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| - | :*Plac+RBS+CinI+LVA
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| - | ::C7,C8 -> OK
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| - | :*BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007):Positive control -> OK
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| - | |}
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| - |
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| - |
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| - | :--->(3/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
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| - |
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| - |
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| - |
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| - |
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| - | :(1/9)--->OD測
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| - |
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| - |
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| - |
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| - | '''[[Team:Chiba/protocol/transformation|Transformation]]'''
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| - | :Competent cells : XL10G 30μL
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| - | ::*C0161(2007) [http://partsregistry.org/Part:BBa_C0161]
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| - | ::*C0161(2006) [http://partsregistry.org/Part:BBa_C0161]
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| - | ::*C0261(2007) [http://partsregistry.org/Part:BBa_C0261]
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| - | ::*C0261(2006) [http://partsregistry.org/Part:BBa_C0261]
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| - |
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| - | :--->(4/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
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| - |
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| - | ===3 September, 2008===
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| - | :(2/9)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
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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-0903.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 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>
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| - | :From left;
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| - | ::*S03154 -> OK
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| - | ::*R1,R3~R7 (Plac+RBS+RhlI+LVA) -> OK
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| - | |}
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| - |
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| - | {|align="justify"
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| - | |[[Image:Chiba-0903-2.JPG]]
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| - | |
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| - | :From left;
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| - | ::*C1,C2,C4~C8 (Plac+RBS+CinI+LVA) -> OK
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| - | |}
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| - |
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| - | :--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
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| - | ::*R1,R3~R7 (Plac+RBS+RhlI+LVA)
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| - | ::*C1,C2,C4~C8 (Plac+RBS+CinI+LVA)
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| - |
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| - | <table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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| - | <tr>
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| - | <td width="257">Digestion</td>
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| - | <td>Single</td><td>Double</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><td>3</td>
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| - | </tr>
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| - | <tr>
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| - | <td>XbaⅠ</td>
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| - | <td>0.1</td><td>0.1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>PstⅠ</td>
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| - | <td>0.1</td><td>0.1</td>
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| - | </tr>
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| - | <tr>
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| - | <td>Buffer 2</td>
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| - | <td>0.9</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>-0.8</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>
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| - | </tr>
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| - | <tr>
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| - | <td>dH2O</td>
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| - | <td>7</td><td>5</td></tr>
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| - | <tr>
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| - | <td>TOTAL</td>
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| - | <td>10</td><td>10</td>
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| - | </tr>
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| - | </table>
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| - |
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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-0903-3.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 DNA</td>
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| - | <td>10</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><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>
| |
| - | :From left;
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| - | :*Single Digestion : R1,R3~R7 -> OK
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| - | :*Single Digestion : C1,C2,C4~C8 -> OK
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| - | |}
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| - |
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| - | {|align="justify"
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| - | |[[Image:Chiba-0903-4.JPG]]
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| - | |
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| - | :From left;
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| - | :*Double Digestion : R1,R3~R7 -> OK
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| - | :*Double Digestion : C1,C2,C4~C8 -> OK
| |
| - | |}
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| - |
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| - |
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| - | (2/9)--->'''Phenotype-test : mix'''
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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>
| |
| - | <td>1</td><td>2</td><td>3</td><td>4</td><td>5</td><td>6</td><td>7</td><td>8</td><td>9</td><td>10</td><td>11</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>L1</td>
| |
| - | <td>2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <tr><td>L2</td><td>-</td><td>2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>L3</td><td>-</td><td>-</td><td>2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>L4</td><td>-</td><td>-</td><td>-</td><td>2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>R1</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>R2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>R3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>R4</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - |
| |
| - | <tr><td>BBa_S03154[http://partsregistry.org/Part:BBa_S03154]</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>2</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>BBa_T9002[http://partsregistry.org/Part:BBa_T9002]</td><td>2</td><td>2</td><td>2</td><td>2</td><td>1</td><td>1</td><td>1</td><td>1</td><td>2</td><td>1</td><td>1</td></tr>
| |
| - | <tr><td>AHL</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
| |
| - | </table>
| |
| - | :::|
| |
| - | :::| 8h
| |
| - | :::V
| |
| - | ::Spindown
| |
| - | :::|
| |
| - | :::|
| |
| - | :::V
| |
| - | ::the intensity GFP
| |
| - |
| |
| - | ===4 September, 2008===
| |
| - |
| |
| - | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0904.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><td>④</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>3</td><td>3</td><td>3</td><td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>2</td><td>2</td><td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>7</td><td>7</td><td>7</td><td>7</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td><td>12</td><td>12</td><td>12</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | :From left;
| |
| - | :
| |
| - | :
| |
| - | |}
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0904-2.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><td>④</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>3</td><td>3</td><td>3</td><td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>2</td><td>2</td><td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>7</td><td>7</td><td>7</td><td>7</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td><td>12</td><td>12</td><td>12</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | :From left;
| |
| - | :
| |
| - | :
| |
| - | |}
| |
| - |
| |
| - | {|align="justify"
| |
| - | |[[Image:Chiba-0904-3.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><td>④</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Sample DNA</td>
| |
| - | <td>3</td><td>3</td><td>3</td><td>3</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>Loading Dye</td>
| |
| - | <td>2</td><td>2</td><td>2</td><td>2</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>dH2O</td>
| |
| - | <td>7</td><td>7</td><td>7</td><td>7</td>
| |
| - | </tr>
| |
| - | <tr>
| |
| - | <td>TOTAL</td>
| |
| - | <td>12</td><td>12</td><td>12</td><td>12</td>
| |
| - | </tr>
| |
| - | </table>
| |
| - | :From left;
| |
| - | :
| |
| - | :
| |
| - | |}
| |
| - |
| |
| - | ===5 September, 2008===
| |
| - |
| |
| - | ===6 September, 2008===
| |
| - |
| |
| - |
| |
| - | {| 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]]
| |
| - | |}
| |
"
