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- | '''入力班のぺーじ。'''
| + | <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> |
| + | >[[Team:Chiba/Internal|ノートに戻る]] |
| | | |
| + | '''入力班のページ''' |
| | | |
- | == 2008.8.20==
| + | [[Team:Chiba/notebook/input/august|8月の実験ログ]] |
- | PCR:'''[[Team:Chiba/protocol/PCR|PCR]]'''
| + | /[[Team:Chiba/notebook/input/september|9月の実験ログ]] |
- | <BR>・BBa_J22136(2007)①
| + | |
- | <BR>・BBa_J22136(2008)②
| + | |
- | <BR>・BBa_J22141(2007)③
| + | |
- | <BR>・BBa_J22141(2008)④
| + | |
- | <BR>・BBa_E0612(2008)⑤
| + | |
- | <table width="250" border="4" 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>DNA tamplate</td>
| + | |
- | <td>1</td><td>1</td><td>1</td><td>1</td>><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>FW primer</td>
| + | |
- | <td>5</td><td>5</td><td>5</td><td>5</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>VR primer</td>
| + | |
- | <td>5</td><td>5</td><td>5</td><td>5</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dNTPmix</td>
| + | |
- | <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>thermo pol buffer</td>
| + | |
- | <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>DNA pol(VENT)</td>
| + | |
- | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>68</td><td>68</td><td>68</td><td>68</td><td>68</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>100</td><td>100</td><td>100</td><td>100</td><td>100</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | <BR>→gel check:'''[[Team:Chiba/protocol/gel check|Gel Check]]'''
| + | |
- | <table width="200" border="4" 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>Sample DNA</td>
| + | |
- | <td>1</td><td>1</td><td>1</td><td>1</td>><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>loading dye</td>
| + | |
- | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH<sub>2</sub>O</td>
| + | |
- | <td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Total</td>
| + | |
- | <td>6</td><td>6</td><td>6</td><td>6</td><td>6</td>
| + | |
- | </tr>
| + | |
| | | |
- | </table>
| |
- | →どれもバンド出ず。
| |
| | | |
| | | |
| | | |
- | | + | {| style="color:white;" cellpadding="3" cellspacing="3" border="0" width="100%" align="center" class="menu" | |
- | <BR>trnsformation:'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| + | |
- | | + | |
- | <BR>・BBa_R0051(2007)
| + | |
- | <BR>・BBa_R0051(2006)
| + | |
- | <BR>・BBa_J06650(2007)
| + | |
- | <BR>・BBa_J06650(2006)
| + | |
- | <BR>・BBa_J22136(2007)
| + | |
- | <BR>・BBa_J22141(2007)
| + | |
- | | + | |
- | →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)
| + | |
- | | + | |
- | == 2008.8.21 ==
| + | |
- | 8.20にtransformationしたもの
| + | |
- | <BR>・BBa_R0051(2007)
| + | |
- | <BR>・BBa_R0051(2006)
| + | |
- | <BR>・BBa_J06650(2007)
| + | |
- | <BR>・BBa_J06650(2006)
| + | |
- | <BR>・BBa_J22136(2007)
| + | |
- | <BR>・BBa_J22141(2007)
| + | |
- | 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、
| + | |
- | さらに1h培養した。
| + | |
- | | + | |
- | == 2008.8.22 ==
| + | |
- | mini prep:'''[[Team:Chiba/protocol/mini prep|Mini Prep]]'''
| + | |
- | <BR>2ml培養した
| + | |
- | <BR>・BBa_R0051(2007)
| + | |
- | <BR>・BBa_R0051(2006)
| + | |
- | <BR>・BBa_J06650(2007)
| + | |
- | <BR>・BBa_J06650(2006)
| + | |
- | <BR>・BBa_J22136(2007)
| + | |
- | <BR>・BBa_J22141(2007)
| + | |
- | <BR>をミニプレした。
| + | |
- | | + | |
- | <BR>insert check:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
| + | |
- | | + | |
- | <BR>BBa_J06650(2006)
| + | |
- | <BR>BBa_J06650(2007)
| + | |
- | <BR>BBa_J22136(2007)
| + | |
- | <BR>BBa_J22141(2007)
| + | |
- | <BR>BBa_R0051(2006)
| + | |
- | <BR>BBa_R0051(2007)
| + | |
- | <BR>BBa_R0051(2007)とBBa_R0051(2006)はsingleのみ、ほかはsingleとdoubleの両方。
| + | |
- | <BR>
| + | |
- | <BR>混ぜ表
| + | |
- | <table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <td width="257"></td>
| + | |
- | <td>double</td><td>simgle</td>
| + | |
- | <tr>
| + | |
- | <td>dH<sub>2</sub>O</td>
| + | |
- | <td>14</td><td>25</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>10×BSA</td>
| + | |
- | <td>7</td><td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>10×NE</td>
| + | |
- | <td>7</td><td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRI</td>
| + | |
- | <td>3.5</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>PstI</td>
| + | |
- | <td>3.5</td><td>-</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | == 2008.8.25 ==
| + | |
- | BBa_J22136(insert check済み)のグリセロールストックづくり
| + | |
- | <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)
| + | |
- | <BR>培養した試験管から180μl,LB-Ampを880μlとって、OD=0.495にした。
| + | |
- | <BR><table width="210" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <td width="257">OD=0.495培養液</td>
| + | |
- | <td>0.67μl</td>
| + | |
- | <tr>
| + | |
- | <td>60%グリセロール</td>
| + | |
- | <td>0.33μl</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>20%グリセロールストック(計)</td>
| + | |
- | <td>1.00ml</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | == 2008.8.28 ==
| + | |
- | transformation:'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| + | |
- | <BR>BBa_I7106(2007)
| + | |
- | 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]]'''
| + | |
- | <BR>→Digestion:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
| + | |
- | <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>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>10×BSA</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>10×NE</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRI</td>
| + | |
- | <td>0.5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>PstI</td>
| + | |
- | <td>0.5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>DNA</td>
| + | |
- | <td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | Digestion 37℃、30分
| + | |
- | <BR>プラスミドの濃度は33.6ng/μl
| + | |
- | | + | |
- | | + | |
- | | + | |
- | <BR>PCR:'''[[Team:Chiba/protocol/PCR|PCR]]'''
| + | |
- | <BR>RBS+CI
| + | |
- | <BR>混ぜ表
| + | |
- | <table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>DNA</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>FW primer</td>
| + | |
- | <td>2.5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>VR primer</td>
| + | |
- | <td>2.5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dNTPmix</td>
| + | |
- | <td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>thermo pol buffer</td>
| + | |
- | <td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>DNA pol(VENT)</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>34</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>50</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | <BR>ゲルチェック:'''[[Team:Chiba/protocol/Gel Check|Gel Check]]'''
| + | |
- | | + | |
- | | + | |
- | | + | |
- | <table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <td width="100">PCR産物</td>
| + | |
- | <td>1</td>
| + | |
- | <tr>
| + | |
- | <td>Dye</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH<SUB>2</SUB>O</td>
| + | |
- | <td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | →濃度は33.6ng/μg
| + | |
- | | + | |
- | <BR>ベクターのPCR:'''[[Team:Chiba/protocol/PCR|PCR]]'''
| + | |
- | <BR>BBa_I7106
| + | |
- | <BR>混ぜ表
| + | |
- | <table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>DNA</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>FW primer</td>
| + | |
- | <td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>VR primer</td>
| + | |
- | <td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dNTPmix</td>
| + | |
- | <td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>thermo pol buffer</td>
| + | |
- | <td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>DNA pol(VENT)</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>68</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>100</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | <BR>ゲルチェック:'''[[Team:Chiba/protocol/Gel Check|Gel Check]]'''
| + | |
- | | + | |
- | | + | |
- | | + | |
- | <table width="150" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <td width="100">PCR産物</td>
| + | |
- | <td>1</td>
| + | |
- | <tr>
| + | |
- | <td>Dye</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH<SUB>2</SUB>O</td>
| + | |
- | <td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | →バンド出ず。
| + | |
- | | + | |
- | == 2008.8.30 ==
| + | |
- | <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>
| + | |
- | →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)
| + | |
- | | + | |
- | {| 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]] |
| + | |} |