Team:Chiba/jk/α

From 2008.igem.org

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'''入力班のぺーじ。'''
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<html><link rel="stylesheet" href="https://2008.igem.org/wiki/index.php?title=User:Maiko/chiba.css&action=raw&ctype=text/css" type="text/css" /></html>
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>[[Team:Chiba/Internal|ノートに戻る]]
 +
'''入力班のページ'''
-
== 2008.8.20==
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[[Team:Chiba/notebook/input/august|８月の実験ログ]]
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PCR:'''[[Team:Chiba/protocol/PCR|PCR]]'''
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/[[Team:Chiba/notebook/input/september|９月の実験ログ]]
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<BR>・BBa_J22136(2007)①
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-
<BR>・BBa_J22136(2008)②
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<BR>・BBa_J22141(2007)③
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-
<BR>・BBa_J22141(2008)④
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-
<BR>・BBa_E0612(2008)⑤
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<table width="250" border="4" 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><td>⑤</td>
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</tr>
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<tr>
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<td>DNA tamplate</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>FW primer</td>
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<td>5</td><td>5</td><td>5</td><td>5</td><td>5</td>
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</tr>
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<tr>
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<td>VR primer</td>
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<td>5</td><td>5</td><td>5</td><td>5</td><td>5</td>
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</tr>
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<tr>
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<td>dNTPmix</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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<tr>
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<td>thermo pol buffer</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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<tr>
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<td>DNA pol(VENT)</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>68</td><td>68</td><td>68</td><td>68</td><td>68</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>100</td><td>100</td><td>100</td>
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</tr>
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</table>
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-
<BR>→gel check:'''[[Team:Chiba/protocol/gel check|Gel Check]]'''
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-
<table width="200" border="4" 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><td>⑤</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>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>loading dye</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>dH<sub>2</sub>O</td>
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<td>4</td><td>4</td><td>4</td><td>4</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>6</td><td>6</td><td>6</td><td>6</td><td>6</td>
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</tr>
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-
</table>
 
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→どれもバンド出ず。
 
-
 
+
{| style="color:white;" cellpadding="3" cellspacing="3" border="0" width="100%" align="center" class="menu" |
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<BR>trnsformation:'''[[Team:Chiba/protocol/transformation|Transformation]]'''
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-
 
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<BR>・BBa_R0051(2007)
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<BR>・BBa_R0051(2006)
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<BR>・BBa_J06650(2007)
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<BR>・BBa_J06650(2006)
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<BR>・BBa_J22136(2007)
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<BR>・BBa_J22141(2007)
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-
 
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→BBa_J22141のみコロニーがほとんど生えていなかった。(１個のみ)
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-
 
+
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== 2008.8.21 ==
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-
8.20にtransformationしたもの
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<BR>・BBa_R0051(2007)
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<BR>・BBa_R0051(2006)
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<BR>・BBa_J06650(2007)
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<BR>・BBa_J06650(2006)
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<BR>・BBa_J22136(2007)
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<BR>・BBa_J22141(2007)
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-
を２ml培養、12h。ただしコロニーが１個だけだったBBa_J22141は2ml培養の11h後にIPTGを20μl加え、
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-
さらに1h培養した。
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-
 
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-
== 2008.8.22 ==
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-
mini prep:'''[[Team:Chiba/protocol/mini prep|Mini Prep]]'''
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<BR>2ml培養した
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<BR>・BBa_R0051(2007)
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<BR>・BBa_R0051(2006)
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<BR>・BBa_J06650(2007)
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<BR>・BBa_J06650(2006)
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<BR>・BBa_J22136(2007)
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<BR>・BBa_J22141(2007)
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<BR>をミニプレした。
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-
 
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<BR>insert check:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
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-
 
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<BR>BBa_J06650(2006)
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<BR>BBa_J06650(2007)
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<BR>BBa_J22136(2007)
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<BR>BBa_J22141(2007)
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<BR>BBa_R0051(2006)
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<BR>BBa_R0051(2007)
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<BR>BBa_R0051(2007)とBBa_R0051(2006)はsingleのみ、ほかはsingleとdoubleの両方。
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-
<BR>
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<BR>混ぜ表
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<table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<td width="257"></td>
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<td>double</td><td>simgle</td>
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<tr>
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<td>dH<sub>2</sub>O</td>
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<td>14</td><td>25</td>
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</tr>
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<tr>
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<td>10×BSA</td>
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<td>7</td><td>10</td>
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</tr>
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<tr>
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<td>10×NE</td>
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<td>7</td><td>10</td>
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-
</tr>
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<tr>
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<td>EcoRI</td>
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<td>3.5</td><td>5</td>
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-
</tr>
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<tr>
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<td>PstI</td>
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<td>3.5</td><td>-</td>
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</tr>
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-
</table>
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-
 
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== 2008.8.25 ==
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BBa_J22136(insert check済み)のグリセロールストックづくり
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<BR><BR>プレートに生えたコロニーをpickして、試験管で2ml培養(LB-Amp,37℃,12h)
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<BR>培養した試験管から180μl,LB-Ampを880μlとって、OD=0.495にした。
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<BR><table width="210" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<td width="257">OD=0.495培養液</td>
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<td>0.67μl</td>
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<tr>
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<td>60%グリセロール</td>
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<td>0.33μl</td>
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</tr>
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<tr>
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<td>20%グリセロールストック(計)</td>
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<td>1.00ml</td>
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</tr>
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-
</table>
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-
 
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== 2008.8.28 ==
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transformation:'''[[Team:Chiba/protocol/transformation|Transformation]]'''
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<BR>BBa_I7106(2007)
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生えてきたコロニーを10ml培養してmini prep(60mlで溶出):'''[[Team:Chiba/protocol/mini prep|mini Prep]]'''
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-
<BR>→Digestion:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
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<BR>混ぜ表
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<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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-
<td width="257"></td>
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<td>double</td>
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<tr>
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<td>dH<sub>2</sub>O</td>
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<td>2</td>
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</tr>
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<tr>
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<td>10×BSA</td>
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<td>1</td>
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-
</tr>
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<tr>
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<td>10×NE</td>
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<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>
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</tr>
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<tr>
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<td>PstI</td>
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<td>0.5</td>
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</tr>
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<tr>
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<td>DNA</td>
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<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>
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-
</tr>
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-
</table>
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-
Digestion 37℃、30分
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<BR>プラスミドの濃度は33.6ng/μl
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-
 
+
-
 
+
-
 
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<BR>PCR:'''[[Team:Chiba/protocol/PCR|PCR]]'''
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<BR>RBS＋CI
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<BR>混ぜ表
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-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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-
<tr>
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-
<td width="257">
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-
</tr>
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<tr>
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<td>DNA</td>
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<td>1</td>
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</tr>
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<tr>
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<td>FW primer</td>
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<td>2.5</td>
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-
</tr>
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-
<tr>
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<td>VR primer</td>
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<td>2.5</td>
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-
</tr>
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<tr>
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<td>dNTPmix</td>
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<td>5</td>
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-
</tr>
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<tr>
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<td>thermo pol buffer</td>
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<td>5</td>
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-
</tr>
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<tr>
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<td>DNA pol(VENT)</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>34</td>
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</tr>
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<tr>
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<td>TOTAL</td>
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<td>50</td>
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-
</tr>
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-
</table>
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-
 
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<BR>ゲルチェック:'''[[Team:Chiba/protocol/Gel Check|Gel Check]]'''
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-
 
+
-
 
+
-
 
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<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<td width="100">PCR産物</td>
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<td>1</td>
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<tr>
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<td>Dye</td>
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<td>1</td>
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-
</tr>
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-
<tr>
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<td>dH<SUB>2</SUB>O</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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-
 
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→濃度は33.6ng/μg
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-
 
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<BR>ベクターのPCR:'''[[Team:Chiba/protocol/PCR|PCR]]'''
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<BR>BBa_I7106
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<BR>混ぜ表
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-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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-
<tr>
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<td width="257">
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-
</tr>
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<tr>
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<td>DNA</td>
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-
<td>1</td>
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-
</tr>
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<tr>
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<td>FW primer</td>
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-
<td>5</td>
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-
</tr>
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<tr>
+
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<td>VR primer</td>
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<td>5</td>
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-
</tr>
+
-
<tr>
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<td>dNTPmix</td>
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-
<td>10</td>
+
-
</tr>
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-
<tr>
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<td>thermo pol buffer</td>
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-
<td>10</td>
+
-
</tr>
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-
<tr>
+
-
<td>DNA pol(VENT)</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>68</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>
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-
</tr>
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-
</table>
+
-
 
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<BR>ゲルチェック:'''[[Team:Chiba/protocol/Gel Check|Gel Check]]'''
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-
 
+
-
 
+
-
 
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<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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-
<td width="100">PCR産物</td>
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-
<td>1</td>
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-
<tr>
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-
<td>Dye</td>
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-
<td>1</td>
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-
</tr>
+
-
<tr>
+
-
<td>dH<SUB>2</SUB>O</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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-
→バンド出ず。
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-
 
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-
== 2008.8.30 ==
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<BR>Digestion:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
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-
<BR>x-RBS-cI-s-p(780bp)
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<BR>混ぜ表
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-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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-
<td width="257"></td>
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-
<td>double</td>
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-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
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-
<td>6</td>
+
-
</tr>
+
-
<tr>
+
-
<td>10×BSA</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>10×NE</td>
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-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>PstI</td>
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-
<td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>XbaI</td>
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-
<td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>DNA</td>
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-
<td>33.6ng/μl×30μl(1μg)</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>60</td>
+
-
</tr>
+
-
</table>
+
-
→30μlゲル抽出
+
-
<BR>→ゲル抽出:'''[[Team:Chiba/protocol/gel extraction|Gel Extraction]]'''
+
-
<BR>→Zymo:'''[[Team:Chiba/protocol/zymo|Zymo]]'''
+
-
<BR>5μ抽出,うち1μlをゲルチェック→OK,4μlはLigation用
+
-
<BR>
+
-
<BR>Digestion:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
+
-
<BR>-e-x-Ptet-s-p-(PMB1,Amp)(Insert:54bp,Vector:2079bp)
+
-
<BR>混ぜ表
+
-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="257"></td>
+
-
<td>double</td>
+
-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
+
-
<td>4</td>
+
-
</tr>
+
-
<tr>
+
-
<td>10×BSA</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>10×NE</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>PstI</td>
+
-
<td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>SpeI</td>
+
-
<td>4</td>
+
-
</tr>
+
-
<tr>
+
-
<td>DNA</td>
+
-
<td>75ng/μl×30μl(2μg)</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>60</td>
+
-
</tr>
+
-
</table>
+
-
→30μlゲル抽出
+
-
<BR>ゲル抽出:'''[[Team:Chiba/protocol/gel extraction|Gel Extraction]]'''
+
-
<BR>→Zymo:'''[[Team:Chiba/protocol/zymo|Zymo]]'''
+
-
<BR>→SAP:'''[[Team:Chiba/protocol/SAP|SAP]]'''
+
-
<BR>混ぜ表
+
-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="57"></td>
+
-
<td></td>
+
-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
+
-
<td>9</td>
+
-
</tr>
+
-
<tr>
+
-
<td>DNA</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>SAP</td>
+
-
<td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>SAP Buffer</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>30</td>
+
-
</tr>
+
-
</table>
+
-
5μ抽出,うち1μlをゲルチェック→OK,4μlはLigation用
+
-
 
+
-
<BR>Ligation:'''[[Team:Chiba/protocol/Ligation|Ligation]]'''
+
-
<BR>混ぜ表
+
-
<table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="257"></td>
+
-
<td></td><td>b.g</td>
+
-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
+
-
<td>1.4</td><td>8</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Ligase</td>
+
-
<td>1</td><td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Ligase Buffer</td>
+
-
<td>2</td><td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Vector</td>
+
-
<td>1.8</td><td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Insert</td>
+
-
<td>3</td><td>-</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>10</td><td>10</td>
+
-
</tr>
+
-
</table>
+
-
→Transformation(XL10G):'''[[Team:Chiba/protocol/transformation|Transformation]]'''
+
-
<BR>→CFU40(b.gは22)
+
-
 
+
-
<BR>コロニーPCR(16コつつく):'''[[Team:Chiba/protocol/colonyPCR|ColonyPCR]]'''
+
-
<BR>混ぜ表
+
-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<tr>
+
-
<td width="257">
+
-
</tr>
+
-
<tr>
+
-
<td>VF</td>
+
-
<td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>VFR</td>
+
-
<td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>dNTP</td>
+
-
<td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>thermo pol buffer</td>
+
-
<td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Tag</td>
+
-
<td>0.3</td>
+
-
</tr>
+
-
<tr>
+
-
<td>dH2O</td>
+
-
<td>11.7</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>20</td>
+
-
</tr>
+
-
</table>
+
-
→Insertチェックしたら1つだけ正しい位置にバンドが出現。そのコロニーを2ml培養し、mini prepした。
+
-
<BR>→mini prep:'''[[Team:Chiba/protocol/mini prep|Mini Prep]]'''
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
<BR>Digestion:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
+
-
<BR>x-RBS-cI-s-p(780bp)
+
-
<BR>混ぜ表
+
-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="257"></td>
+
-
<td>double</td>
+
-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
+
-
<td>6</td>
+
-
</tr>
+
-
<tr>
+
-
<td>10×BSA</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>10×NE</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>PstI</td>
+
-
<td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>XbaI</td>
+
-
<td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>DNA</td>
+
-
<td>33.6ng/μl×30μl(1μg)</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>60</td>
+
-
</tr>
+
-
</table>
+
-
<BR>ゲル抽出:'''[[Team:Chiba/protocol/gel extraction|Gel Extraction]]'''
+
-
<BR>混ぜ表
+
-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="57"></td>
+
-
<td></td>
+
-
<tr>
+
-
<td>Digestion産物</td>
+
-
<td>30</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Dye</td>
+
-
<td>6</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>36</td>
+
-
</tr>
+
-
</table>
+
-
<BR>→Zymo:'''[[Team:Chiba/protocol/zymo|Zymo]]'''
+
-
<BR>→NFW5μlで溶出
+
-
<BR>ゲルチェック:'''[[Team:Chiba/protocol/gel check|Gel Check]]'''
+
-
<BR>混ぜ表
+
-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="57"></td>
+
-
<td></td>
+
-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
+
-
<td>4</td>
+
-
</tr>
+
-
<tr>
+
-
<td>DNA</td>
+
-
<td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Dye</td>
+
-
<td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>6</td>
+
-
</tr>
+
-
</table>
+
-
→ゲルチェックOK
+
-
<BR>
+
-
<BR>Digestion:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
+
-
<BR>-e-x-Ptet-s-p-(PMB1,Amp)(Insert:54bp,Vector:2079bp)
+
-
<BR>混ぜ表
+
-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="257"></td>
+
-
<td>double</td>
+
-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
+
-
<td>4</td>
+
-
</tr>
+
-
<tr>
+
-
<td>10×BSA</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>10×NE</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>PstI</td>
+
-
<td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>SpeI</td>
+
-
<td>4</td>
+
-
</tr>
+
-
<tr>
+
-
<td>DNA</td>
+
-
<td>75ng/μl×30μl(2.25μg)</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>60</td>
+
-
</tr>
+
-
</table>
+
-
 
+
-
<BR>ゲル抽出:'''[[Team:Chiba/protocol/gel extraction|Gel Extraction]]'''
+
-
<BR>混ぜ表
+
-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="57"></td>
+
-
<td></td>
+
-
<tr>
+
-
<td>Digestion産物</td>
+
-
<td>30</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Dye</td>
+
-
<td>6</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>36</td>
+
-
</tr>
+
-
</table>
+
-
<BR>→Zymo:'''[[Team:Chiba/protocol/zymo|Zymo]]'''
+
-
<BR>→NFW10μlで溶出
+
-
 
+
-
<BR>→SAP:'''[[Team:Chiba/protocol/SAP|SAP]]'''
+
-
<BR>混ぜ表
+
-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="57"></td>
+
-
<td></td>
+
-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
+
-
<td>9</td>
+
-
</tr>
+
-
<tr>
+
-
<td>DNA</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>SAP</td>
+
-
<td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>SAP Buffer(×10)</td>
+
-
<td>10</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>30</td>
+
-
</tr>
+
-
</table>
+
-
→37℃で1h、乾燥機(65℃)で15min,
+
-
<BR>→Binding Bufferを90μl加え、VORTEX
+
-
<BR>→Zymo:'''[[Team:Chiba/protocol/zymo|Zymo]]'''
+
-
→NFW5μlで抽出
+
-
<BR>ゲルチェック:'''[[Team:Chiba/protocol/gel check|Gel Check]]'''
+
-
<BR>混ぜ表
+
-
<table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="57"></td>
+
-
<td></td>
+
-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
+
-
<td>4</td>
+
-
</tr>
+
-
<tr>
+
-
<td>DNA</td>
+
-
<td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Dye</td>
+
-
<td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>6</td>
+
-
</tr>
+
-
</table>
+
-
→ゲルチェックOK
+
-
 
+
-
<BR>Ligation:'''[[Team:Chiba/protocol/Ligation|Ligation]]'''
+
-
<BR>混ぜ表
+
-
<table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="257"></td>
+
-
<td></td><td>b.g</td>
+
-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
+
-
<td>1.4</td><td>8</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Ligase</td>
+
-
<td>1</td><td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Ligase Buffer</td>
+
-
<td>2</td><td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Vector</td>
+
-
<td>1.8</td><td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Insert</td>
+
-
<td>3</td><td>-</td>
+
-
</tr>
+
-
<tr>
+
-
<td>TOTAL</td>
+
-
<td>10</td><td>10</td>
+
-
</tr>
+
-
</table>
+
-
→25℃で2h放置
+
-
→Transformation(XL10G):'''[[Team:Chiba/protocol/transformation|Transformation]]'''
+
-
<BR>→CFU40(b.gは22)
+
-
 
+
-
== 2008.9.1 ==
+
-
・Ptet＋RBS＋cIの、Mini Prep産物の濃度チェック。
+
-
<BR>-Ptet-cI-pMB1-Amp-の機能チェック。
+
-
<BR>-Ptet-cI-pMB1-Amp-,-PcI-GFP-p15A-Cm-をDouble Transformation(BWΔflic)
+
-
<BR>結果→GFPが発現していた。
+
-
<BR>→-PcI-GFP-p15A-Cm-が機能していないのでは？
+
-
<BR>→ターミネーターをつける、pSB1A3にのせかえる。
+
-
 
+
-
<BR>-Ptet-cI-pMB1-Amp-のDigestion Check
+
-
 
+
-
<BR>混ぜ表
+
-
<table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
+
-
<td width="257"></td>
+
-
<td>Double</td><td>Single</td>
+
-
<tr>
+
-
<td>dH<sub>2</sub>O</td>
+
-
<td>1</td><td>2</td>
+
-
</tr>
+
-
<tr>
+
-
<td>XbaI</td>
+
-
<td>1</td><td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>SpeI</td>
+
-
<td>1</td><td>-</td>
+
-
</tr>
+
-
<tr>
+
-
<td>BSA(×10)</td>
+
-
<td>1</td><td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<td>NEB(×10)</td>
+
-
<td>1</td><td>1</td>
+
-
</tr>
+
-
<tr>
+
-
<tr>
+
-
<td>DNA</td>
+
-
<td>5</td><td>5</td>
+
-
</tr>
+
-
<td>TOTAL</td>
+
-
<td>10</td><td>10</td>
+
-
</tr>
+
-
</table>
+
-
 
+
-
 
+
-
 
+
-
{| style="color:green;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/j|ホーム]]
!align="center"|[[Team:Chiba/Team/j|メンバー紹介]]
!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/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/Parts/j|作成したDNAパーツ]]
-
!align="center"|[[Team:Chiba/Modeling/j|モデリング]]
+
!align="center"|[[Team:Chiba/Internal|ノート]]
!align="center"|[[Team:Chiba/Internal|ノート]]
 +
|}

Latest revision as of 11:26, 14 December 2008

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