Difference between revisions of "Team:Paris Bettencourt/Notebook/Riboswitch"
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Gel order : - - - B5 B6 B7 I4 I5 ( I3 was lost )<br> | Gel order : - - - B5 B6 B7 I4 I5 ( I3 was lost )<br> | ||
<img src="https://static.igem.org/mediawiki/2015/c/c9/7687.png" width="300px"> | <img src="https://static.igem.org/mediawiki/2015/c/c9/7687.png" width="300px"> | ||
+ | |||
+ | |||
+ | <h1 class="date three">September 2nd</h1> | ||
+ | |||
+ | <h2>Goal 1</h2> | ||
+ | Digestion/Ligation | ||
+ | |||
+ | <h2>Procedure</h2> | ||
+ | The PCR product was purified before. <br> | ||
+ | <style type="text/css"> | ||
+ | .tg {border-collapse:collapse;border-spacing:0;} | ||
+ | .tg td{font-family:Arial, sans-serif;font-size:14px;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;} | ||
+ | .tg th{font-family:Arial, sans-serif;font-size:14px;font-weight:normal;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;} | ||
+ | </style> | ||
+ | <table class="tg"> | ||
+ | <tr> | ||
+ | <th class="tg-031e"></th> | ||
+ | <th class="tg-031e">F11d</th> | ||
+ | <th class="tg-031e">pSB1C3d</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">DNA 1:3</td> | ||
+ | <td class="tg-031e">55ng/uL, 1.5 ug —> 30 uL</td> | ||
+ | <td class="tg-031e">96 ng/uL, 0.75 ug —> 8 uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">Fast AP</td> | ||
+ | <td class="tg-031e"></td> | ||
+ | <td class="tg-031e">2uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">EcoRI</td> | ||
+ | <td class="tg-031e">2uL</td> | ||
+ | <td class="tg-031e">2uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">SpeI</td> | ||
+ | <td class="tg-031e">2uL</td> | ||
+ | <td class="tg-031e">2uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">FD Buffer</td> | ||
+ | <td class="tg-031e">4uL</td> | ||
+ | <td class="tg-031e">6uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">Water</td> | ||
+ | <td class="tg-031e">2uL</td> | ||
+ | <td class="tg-031e">40uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">TOTAL</td> | ||
+ | <td class="tg-031e">40uL</td> | ||
+ | <td class="tg-031e">60uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">Nanodrop after purif</td> | ||
+ | <td class="tg-031e">8.6</td> | ||
+ | <td class="tg-031e">11 (after gel extract)</td> | ||
+ | </tr> | ||
+ | </table><br> | ||
+ | |||
+ | <img src="https://static.igem.org/mediawiki/2015/a/ab/9877.png" width="300px"><br> | ||
+ | I extracted the 3kb band from the gel | ||
+ | <br> | ||
+ | Ligation : <br> | ||
+ | <style type="text/css"> | ||
+ | .tg {border-collapse:collapse;border-spacing:0;} | ||
+ | .tg td{font-family:Arial, sans-serif;font-size:14px;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;} | ||
+ | .tg th{font-family:Arial, sans-serif;font-size:14px;font-weight:normal;padding:10px 5px;border-style:solid;border-width:1px;overflow:hidden;word-break:normal;} | ||
+ | </style> | ||
+ | <table class="tg"> | ||
+ | <tr> | ||
+ | <th class="tg-031e"></th> | ||
+ | <th class="tg-031e">La (F2d’-LP1d’)</th> | ||
+ | <th class="tg-031e">La c</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">F11d</td> | ||
+ | <td class="tg-031e">25 uL</td> | ||
+ | <td class="tg-031e"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">pSB1C3d</td> | ||
+ | <td class="tg-031e">10uL</td> | ||
+ | <td class="tg-031e">10uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">Ligase</td> | ||
+ | <td class="tg-031e">1uL</td> | ||
+ | <td class="tg-031e">1uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">L buffer</td> | ||
+ | <td class="tg-031e">4uL</td> | ||
+ | <td class="tg-031e">4uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">Water</td> | ||
+ | <td class="tg-031e"></td> | ||
+ | <td class="tg-031e">25uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">TOTAL</td> | ||
+ | <td class="tg-031e">40uL</td> | ||
+ | <td class="tg-031e">40uL</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | Then on the bench for 1 hour |
Revision as of 15:27, 16 September 2015