|
|
| Line 20: |
Line 20: |
| | <article> | | <article> |
| | <h1>Monday, August 29<sup>th</sup></h3> | | <h1>Monday, August 29<sup>th</sup></h3> |
| − | <h2>Tasks:</h2>
| + | |
| − | <div class="row">
| + | |
| − | <div class="col-sm-2">
| + | |
| − | <p>Tasfia</p>
| + | |
| − | </div>
| + | |
| − | <div class="col-sm-10">
| + | |
| − | <ul>
| + | |
| − | <li>Made 5 mL overnight cultures of "TetR-Cas9," "Assembled Cas9 from Cam Culture," GFP from iGEM kit</li>
| + | |
| − | <ul>
| + | |
| − | <li>Three different colonies</li>
| + | |
| − | <li>Used 1000X Cam</li>
| + | |
| − | <li>Working concentration was 35 ug/mL</li>
| + | |
| − | <li>Added 5.15 uL to each culture tube </li>
| + | |
| − | </ul>
| + | |
| − | <li>Made 5 mL overnight culture of gRNA Gibson product</li>
| + | |
| − | <ul>
| + | |
| − | <li>One colony, duplicate overnight cultures</li>
| + | |
| − | <li>Used 1000X Tet (10 mg/mL)</li>
| + | |
| − | <li>Working concentration was 10 ug/mL</li>
| + | |
| − | <li>Added 5 uL to each culture tube</li>
| + | |
| − | </ul>
| + | |
| − | <li>Gel extracted signal sequences (with homology added)
| + | |
| − | <div class="row">
| + | |
| − | <div class="col-xs-8 col-sm-12"><img src="https://static.igem.org/mediawiki/2016/5/50/T--Northwestern--08_28_1.png" width="1210" height="440" alt=""/></div>
| + | |
| − | </div>
| + | |
| − | </li>
| + | |
| − | <li>Used team-modified protocol:</li>
| + | |
| − | <ul>
| + | |
| − | <li>Sat product for ~4 minutes between washes</li>
| + | |
| − | <li>Evaporated EtOH for ~12 minutes</li>
| + | |
| − | <li>Eluted in nuclease-free water after standing for ~15 minutes</li>
| + | |
| − | </ul>
| + | |
| − | <li>Going forward: we can attempt a Cas9-SS Gibson after figuring out what’s wrong with the gRNA Gibson assembly</li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | <div class="row">
| + | |
| − | <div class="col-sm-2">
| + | |
| − | <p>Tyler</p>
| + | |
| − | </div>
| + | |
| − | <div class="col-sm-10">
| + | |
| − | <ul>
| + | |
| − | <li>Retransformed gRNA Gibson product from 08.26.16 </li>
| + | |
| − | <ul>
| + | |
| − | <li>3 µL of gRNA Gibson assembly product</li>
| + | |
| − | <li>2 µL positive Gibson kit control</li>
| + | |
| − | <li>2 µL (-) control 1 µL transformation control</li>
| + | |
| − | </ul>
| + | |
| − | <li>Retransformed 1 µL GFP from iGEM kit</li>
| + | |
| − | </ul>
| + | |
| − | </div>
| + | |
| − | </div>
| + | |
| − | </article>
| + | |
| | <footer id="nav"> | | <footer id="nav"> |
| | <div class="row"> | | <div class="row"> |
