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Team:Chiba/jk/α
From 2008.igem.org
(→2008.8.22) |
(→2008.8.22) |
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master mix混ぜ表 | master mix混ぜ表 | ||
| + | <table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
| + | <tr> | ||
| + | <td width="257"></td> | ||
| + | <td>double</td><td>simgle</td> | ||
| + | </tr> | ||
| + | <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> | ||
| + | <td>PstI</td> | ||
| + | <td>3.5</td><td>-</td> | ||
| + | </tr> | ||
| + | |||
| + | </table> | ||
Revision as of 12:49, 4 September 2008
入力班のぺーじ。
2008.8.20
PCR:PCR
・BBa_J22136(2007)①
・BBa_J22136(2008)②
・BBa_J22141(2007)③
・BBa_J22141(2008)④
・BBa_E0612(2008)⑤
| Sample No | ① | ② | ③ | ④ | ⑤ |
| DNA tamplate | 1 | 1 | 1 | 1 | >1 |
| FW primer | 5 | 5 | 5 | 5 | 5 |
| VR primer | 5 | 5 | 5 | 5 | 5 |
| dNTPmix | 10 | 10 | 10 | 10 | 10 |
| thermo pol buffer | 10 | 10 | 10 | 10 | 10 |
| DNA pol(VENT) | 1 | 1 | 1 | 1 | 1 |
| dH2O | 68 | 68 | 68 | 68 | 68 |
| TOTAL | 100 | 100 | 100 | 100 | 100 |
→gel check:Gel Check
| Sample No | ① | ② | ③ | ④ | ⑤ |
| Sample DNA | 1 | 1 | 1 | 1 | >1 |
| loading dye | 1 | 1 | 1 | 1 | 1 |
| dH2O | 4 | 4 | 4 | 4 | 4 |
| Total | 6 | 6 | 6 | 6 | 6 |
→どれもバンド出ず。
trnsformation:Transformation
・BBa_R0051(2007)
・BBa_R0051(2006)
・BBa_J06650(2007)
・BBa_J06650(2006)
・BBa_J22136(2007)
・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
・BBa_R0051(2007)
・BBa_R0051(2006)
・BBa_J06650(2007)
・BBa_J06650(2006)
・BBa_J22136(2007)
・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:Mini Prep
2ml培養した
・BBa_R0051(2007)
・BBa_R0051(2006)
・BBa_J06650(2007)
・BBa_J06650(2006)
・BBa_J22136(2007)
・BBa_J22141(2007)
をミニプレした。
insert check:Digestion
BBa_J06650(2006)
BBa_J06650(2007)
BBa_J22136(2007)
BBa_J22141(2007)
BBa_R0051(2006)
BBa_R0051(2007)
BBa_R0051(2007)とBBa_R0051(2006)はsingleのみ、ほかはsingleとdoubleの両方。
master mix混ぜ表
| double | simgle | |
| dH2O | 14 | 25 |
| 10×BSA | 7 | 10 |
| 10×NE | 7 | 10 |
| EcoRI | 3.5 | 5 | PstI | 3.5 | - |
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