Difference between revisions of "Team:Paris Bettencourt/Notebook/Riboswitch"
Line 669: | Line 669: | ||
<img src="https://static.igem.org/mediawiki/2015/7/7e/555666.png" width="300px"> | <img src="https://static.igem.org/mediawiki/2015/7/7e/555666.png" width="300px"> | ||
− | <br | + | <br> |
Then I cuted the 2100 fragment and did the gel extraction protocol | Then I cuted the 2100 fragment and did the gel extraction protocol | ||
<h2>Goal 3 </h2> | <h2>Goal 3 </h2> | ||
Digestion of pSB1C3 linearized (LP1,2,3 *) and F1,2,3 * | Digestion of pSB1C3 linearized (LP1,2,3 *) and F1,2,3 * | ||
+ | |||
+ | <h2>Procedure</h2> | ||
+ | <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">F2d</th> | ||
+ | <th class="tg-031e">LP2d</th> | ||
+ | <th class="tg-031e">LP3d</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">ADN</td> | ||
+ | <td class="tg-031e">10 uL</td> | ||
+ | <td class="tg-031e">10 uL</td> | ||
+ | <td class="tg-031e">10 uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">EcorRI</td> | ||
+ | <td class="tg-031e">2 uL</td> | ||
+ | <td class="tg-031e">2 uL</td> | ||
+ | <td class="tg-031e">2 uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">PstI</td> | ||
+ | <td class="tg-031e">2 uL</td> | ||
+ | <td class="tg-031e">2 uL</td> | ||
+ | <td class="tg-031e">2 uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">Buffer</td> | ||
+ | <td class="tg-031e">4 uL</td> | ||
+ | <td class="tg-031e">6 uL</td> | ||
+ | <td class="tg-031e">6 uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">Water</td> | ||
+ | <td class="tg-031e">22 uL</td> | ||
+ | <td class="tg-031e">38 uL</td> | ||
+ | <td class="tg-031e">38 uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">FastAP</td> | ||
+ | <td class="tg-031e">_</td> | ||
+ | <td class="tg-031e">2uL</td> | ||
+ | <td class="tg-031e">2uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">TOTAL</td> | ||
+ | <td class="tg-031e">40</td> | ||
+ | <td class="tg-031e">60</td> | ||
+ | <td class="tg-031e">60</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | Then 30min at 37°C | ||
+ | |||
+ | <h2>Goal 4 </h2> | ||
+ | Ligations and transformations | ||
+ | |||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
+ | <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">L1</th> | ||
+ | <th class="tg-031e">L2</th> | ||
+ | <th class="tg-031e">L3</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">Vector 100ng</td> | ||
+ | <td class="tg-031e">LP2d (40.1ng/uL),2uL</td> | ||
+ | <td class="tg-031e">LP3d (21.1ng/uL),4 uL</td> | ||
+ | <td class="tg-031e">Backbone from digestion + electrophoresis extraction (9.1ng/uL), 10 uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">Insert 300 ng (8.1ng/uL) (F2d)</td> | ||
+ | <td class="tg-031e">30 uL</td> | ||
+ | <td class="tg-031e">20 uL</td> | ||
+ | <td class="tg-031e">10 uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">T4 ligase buffer</td> | ||
+ | <td class="tg-031e">4 uL</td> | ||
+ | <td class="tg-031e">4 uL</td> | ||
+ | <td class="tg-031e">2 uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">T4 ligase</td> | ||
+ | <td class="tg-031e">2 uL</td> | ||
+ | <td class="tg-031e">2 uL</td> | ||
+ | <td class="tg-031e">1 uL</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">Water</td> | ||
+ | <td class="tg-031e">0</td> | ||
+ | <td class="tg-031e">10 uL</td> | ||
+ | <td class="tg-031e">0</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-031e">TOTAL</td> | ||
+ | <td class="tg-031e">40 uL</td> | ||
+ | <td class="tg-031e">40 uL</td> | ||
+ | <td class="tg-031e">20 uL</td> | ||
+ | </tr> | ||
+ | </table> |
Revision as of 14:10, 10 September 2015