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| - | {{:Team:Marburg/Template:Header}}
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| - | {{:Team:Marburg/Template:ContentTitle}}
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| - | Notebook
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| - | {{:Team:Marburg/Template:ContentStart}}
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| | | | |
| - | <html>
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| - |
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| - | <!-- Beispiel Allgemein -->
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| - |
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| - | <div class="notebooky-entry">
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| - | <h2 class="title">
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| - | <a name="11-11-2012">11.11.2011</a>
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| - | </h2>
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| - | <fieldset class="experiment general">
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| - | <legend><a name="title">Überschrift</a></legend>
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| - | <div class="investigator">
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| - | <span class="inv">Investigator:</span>
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| - | <span class="inv-names">X,Y</span>
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| - | </div>
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| - | <div class="aim">
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| - | <span class="aim">Aim:</span>
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| - | <span class="aim-desc">We attempt to get results.</span>
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| - | </div>
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| - | <div class="exp-content">
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| - | <p>Wichtig ist, dass immer die verwendete Materialien (Plasmide, genomische DNA [aus welchen Stamm],
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| - | Primer) genau benannt und/oder ggf. irgendwo nummeriert werden. Es sollte alles nachvollziehbar sein,
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| - | wann ihr mit welchen Proben was gemacht habt.</p>
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| - | <p></p>
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| - | <p>Auf den folgenden Seiten sind Beispiele, wie wir uns das Notebook vom Aufbau und Detailgrad vorstellen.</p>
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| - | </div>
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| - | </fieldset>
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| - | </div>
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| - |
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| - |
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| - |
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| - | <!-- Beispiel Ligation, Verdau -->
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| - |
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| - | <div class="notebooky-entry">
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| - | <h2 class="title">
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| - | <a name="17-04-2013">17.04.2013</a>
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| - | </h2>
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| - | <fieldset class="experiment ligation">
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| - | <legend><a name="lig">Ligation</a></legend>
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| - | <div class="investigator">
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| - | <span class="inv">Investigator:</span>
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| - | <span class="inv-names">Franzi, Lucas</span>
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| - | </div>
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| - | <div class="aim">
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| - | <span class="aim">Aim:</span>
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| - | <span class="aim-desc">Assemble the Bricks 1, 3, 4, 5, 6, 7 and 8 into the vector pSB1C3.</span>
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| - | </div>
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| - | <div class="exp-content">
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| - | <ul class="lig">
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| - | <li>5 µl vector DNA (pSB1C3)</li>
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| - | <li>20 µl insert DNA (1, 3, 4, 5, 6, 7 or 8)</li>
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| - | <li>3 µl 10x T4 DNA ligase buffer</li>
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| - | <li>2 µl T4 DNA ligase</li>
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| - | </ul>
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| - | <p>The samples were incubated for 14 h at 18° C.</p>
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| - | </div>
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| - | </fieldset>
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| - | </div>
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| - | <!-- Beispiel Verdau -->
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| - | <fieldset class="experiment digest">
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| - | <legend><a name="dig">Digest</a></legend>
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| - | <div class="investigator">
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| - | <span class="inv">Investigator:</span>
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| - | <span class="inv-names">Christian, Patrick</span>
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| - | </div>
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| - | <div class="aim">
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| - | <span class="aim">Aim:</span>
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| - | <span class="aim-desc">Digest of pSB1C3-J04450 with <i>Mlu</i>I and <i>Hind</i>III.</span>
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| - | </div>
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| - | <div class="exp-content">
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| - | <ul class="digest">
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| - | <li>4 µl DNA (pSB1C3-J04450)</li>
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| - | <li>1 µl MluI</li>
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| - | <li>1 µl HindIII</li>
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| - | <li>1,5 µl 10x red buffer</li>
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| - | <li>1,5 µl 10x orange g loading buffer</li>
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| - | <li>6 µl H2O</li>
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| - | </ul>
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| - | <p>The samples were incubated for 4 h at 37° C.</p>
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| - | <table class="gel digest">
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| - | <colgroup>
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| - | <col width="50%" />
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| - | <col width="50%" />
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| - | </colgroup>
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| - | <thead>
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| - | <tr>
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| - | <th colspan="2" class="title">Gel electrophoresis</th>
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| - | </tr>
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| - | </thead>
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| - | <tbody>
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| - | <tr>
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| - | <td>
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| - | <img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
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| - | </td>
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| - | <td>
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| - | <p>
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| - | <span class="gel-elc">Gel substances</span>
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| - | <ul class="gel-sub">
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| - | <li>1% Agarose gel</li>
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| - | <li>10 µl RedSafe in 50 ml gel</li>
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| - | <li>x µl Hyper Ladder</li>
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| - | </ul>
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| - | </p>
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| - | <p>
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| - | <span class="exp">Expactations</span>
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| - | <ul class="exp">
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| - | <li>Lane 1: 5300 kbp</li>
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| - | <li>Lane 2: 3300 kbp</li>
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| - | <li>Lane 3: 1300 kbp</li>
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| - | </ul>
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| - | </p>
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| - | <p>We don’t receive all expected fragments. The expected fragment in lane 1 is missing.</p>
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| - | </td>
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| - | </tr>
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| - | <tr>
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| - | <td colspan="2" class="gel-fazit">
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| - | <p>We don’t receive all expected fragments. The expected fragment in lane 1 is missing.</p>
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| - | </td>
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| - | </tr>
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| - | </tbody>
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| - | </table>
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| - | </div>
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| - | </fieldset>
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| - |
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| - | <!-- Beispiel Transformation, PCR -->
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| - |
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| - | <div class="notebooky-entry">
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| - | <h2 class="title">
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| - | <a name="18-04-2012">18.04.2012</a>
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| - | </h2>
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| - | <fieldset class="experiment transformation">
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| - | <legend>Transformation</legend>
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| - | <div class="investigator">
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| - | <span class="inv">Investigator:</span>
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| - | <span class="inv-names">Franzi, Christian</span>
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| - | </div>
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| - | <div class="aim">
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| - | <span class="aim">Aim:</span>
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| - | <span class="aim-desc">Transformation of the plasmid DNA pB201-40JO in <i>E. coli</i> for amplification.</span>
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| - | </div>
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| - | <div class="exp-content">
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| - | <p>The chemo competent <i>E. coli</i> DH5a cells were transformed with pB201-40JO DNA and plated on dYT-Amp-plates. <br />The plates were incubated over night at 37° C .</p>
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| - | </div>
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| - | </fieldset>
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| - | <!-- PCR -->
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| - | <fieldset class="experiment pcr">
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| - | <legend><a name="pcr">PCR</a></legend>
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| - | <div class="investigator">
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| - | <span class="inv">Investigator:</span>
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| - | <span class="inv-names">Patrick, Lucas</span>
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| - | </div>
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| - | <div class="aim">
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| - | <span class="aim">Aim:</span>
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| - | <span class="aim-desc">Receive <i>Pvu</i>II point-mutation on colonies 3 and 5 of E. coli pB201-40JO.</span>
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| - | </div>
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| - | <div class="exp-content">
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| - | <table class="pcr">
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| - | <colgroup>
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| - | <col width="15%" />
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| - | <col width="30%" />
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| - | <col width="5%" />
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| - | <col width="20%" />
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| - | <col width="20%" />
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| - | <col width="10%" />
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| - | </colgroup>
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| - | <thead>
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| - | <th>Volume</th>
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| - | <th>Reagent</th>
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| - | <th> </th>
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| - | <th>Temp (°C)</th>
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| - | <th colspan="2">Time</th>
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| - | </thead>
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| - | <tr>
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| - | <td>10 µl</td>
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| - | <td>5x Buffer</td>
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| - | <td> </td>
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| - | <td>95</td>
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| - | <td colspan="2">3 min</td>
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| - | </tr>
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| - | <tr>
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| - | <td>1,5 µl</td>
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| - | <td>Primer fwd 13</td>
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| - | <td> </td>
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| - | <td>95</td>
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| - | <td colspan="2">3 sec</td>
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| - | </tr>
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| - | <tr>
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| - | <td>1,5 µl</td>
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| - | <td>Primer rev 14</td>
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| - | <td> </td>
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| - | <td>58</td>
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| - | <td>30 sec</td>
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| - | <td rowspan="3">x17</td>
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| - | </tr>
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| - | <tr>
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| - | <td>1 µl</td>
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| - | <td>Template</td>
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| - | <td> </td>
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| - | <td>72</td>
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| - | <td>1 min</td>
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| - | </tr>
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| - | <tr>
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| - | <td>36 µl</td>
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| - | <td>H2O</td>
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| - | <td> </td>
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| - | <td>7 min</td>
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| - | <td>3 min</td>
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| - | </tr>
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| - | <tr>
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| - | <td>1 µl Phusion</td>
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| - | <td>Phusion Polymerase</td>
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| - | <td> </td>
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| - | <td>4</td>
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| - | <td colspan="2"><span class="hold">Hold</span></td>
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| - | </tr>
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| - | </table>
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| - | <br />
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| - | <!-- Das Gel-Bild fast gleich wie beim Verdau -->
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| - | <table class="gel pcr">
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| - | <colgroup>
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| - | <col width="50%" />
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| - | <col width="50%" />
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| - | </colgroup>
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| - | <thead>
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| - | <tr>
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| - | <th colspan="2" class="title">Gel electrophoresis</th>
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| - | </tr>
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| - | </thead>
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| - | <tbody>
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| - | <tr>
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| - | <td>
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| - | <img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
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| - | </td>
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| - | <td>
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| - | <p>
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| - | <span class="gel-elc">Gel substances</span>
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| - | <ul class="gel-sub">
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| - | <li>1% Agarose gel</li>
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| - | <li>10 µl RedSafe in 50 ml gel</li>
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| - | <li>x µl Hyper Ladder</li>
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| - | </ul>
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| - | </p>
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| - | <p>
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| - | <span class="exp">Expactations</span>
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| - | <ul class="exp">
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| - | <li>Lane 1: 5300 kbp</li>
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| - | <li>Lane 2: 3300 kbp</li>
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| - | <li>Lane 3: 1300 kbp</li>
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| - | </ul>
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| - | </p>
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| - | <p>We don’t receive all expected fragments. The expected fragment in lane 1 is missing.</p>
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| - | </td>
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| - | </tr>
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| - | </tbody>
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| - | </table>
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| - | </div>
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| - | </fieldset>
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| - |
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| - |
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| - | <!-- Prep Digest -->
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| - | <fieldset class="experiment digest">
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| - | <legend><a name="dig">Digest</a></legend>
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| - | <div class="investigator">
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| - | <span class="inv">Investigator:</span>
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| - | <span class="inv-names">Dominik</span>
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| - | </div>
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| - | <div class="aim">
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| - | <span class="aim">Aim:</span>
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| - | <span class="aim-desc">Digest of pSB1C3-iBB4+iBB11+iBB9, pSB1C3-iBB4+iBB12+iBB9, pSB1C3-iBB6315 and pSB1A3.</span>
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| - | </div>
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| - | <div class="exp-content">
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| - | <p><ul class="digest">
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| - | <li>1000 ng Plasmid (pSB1C3-iBB4+iBB11+iBB9, pSB1C3-iBB4+iBB12+iBB9)</li>
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| - | <li>0.5 µl EcoRI</li>
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| - | <li>0.5 µl SpeI</li>
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| - | <li>2 µl CutSmart</li>
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| - | <li>ad 20 µl ddH2O</li>
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| - | </ul>
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| - |
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| - | <p><ul class="digest">
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| - | <li>1000 ng Plasmid (pSB1C3-iBB6315)</li>
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| - | <li>0.5 µl XbaI</li>
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| - | <li>0.5 µl PstI</li>
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| - | <li>2 µl CutSmart</li>
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| - | <li>ad 20 µl ddH2O</li>
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| - | </ul>
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| - |
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| - | <p><ul class="digest">
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| - | <li>1000 ng Plasmid (pSB1A3)</li>
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| - | <li>0.5 µl EcoRI</li>
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| - | <li>0.5 µl PstI</li>
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| - | <li>2 µl CutSmart</li>
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| - | <li>ad 20 µl ddH2O</li>
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| - | </ul>
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| - |
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| - | <p>The samples were incubated for 1 h at 37° C.</p>
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| - | <table class="gel digest">
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| - | <colgroup>
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| - | <col width="50%" />
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| - | <col width="50%" />
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| - | </colgroup>
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| - | <thead>
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| - | <tr>
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| - | <th colspan="2" class="title">Gel electrophoresis</th>
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| - | </tr>
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| - | </thead>
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| - | <tbody>
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| - | <tr>
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| - | <td>
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| - | <img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
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| - | </td>
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| - | <td>
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| - | <p>
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| - | <span class="gel-elc">Gel substances</span>
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| - | <ul class="gel-sub">
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| - | <li>1% Agarose gel</li>
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| - | <li>10 µl RedSafe in 50 ml gel</li>
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| - | <li>x µl Hyper Ladder</li>
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| - | </ul>
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| - | </p>
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| - | <p>
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| - | <span class="exp">Expactations</span>
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| - | <ul class="exp">
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| - | <li>Lane 1: 5300 kbp</li>
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| - | <li>Lane 2: 3300 kbp</li>
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| - | <li>Lane 3: 1300 kbp</li>
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| - | </ul>
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| - | </p>
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| - | <p>All expected fragments were present. The relevant fragments were cut from the gel and then purified via “Qiagen Gel Extraction Kit” (Qiagen, Düsseldorf).</p>
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| - | </td>
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| - | </tr>
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| - | </tbody>
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| - | </table>
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| - | </div>
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| - | </fieldset>
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| - |
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| - | <!-- Inoculation -->
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| - | <fieldset class="experiment inoculation">
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| - | <legend><a name="ino">Inoculation</a></legend>
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| - | <div class="investigator">
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| - | <span class="inv">Investigator:</span>
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| - | <span class="inv-names">Dominik</span>
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| - | </div>
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| - | <div class="aim">
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| - | <span class="aim">Aim:</span>
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| - | <span class="aim-desc">inoculation of colonies for plasmid preparation</span>
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| - | </div>
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| - | <div class="exp-content">
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| - | <p>4 colonies of pSB1A3-iBB395146 and 4 colonies of pSB1A3-iBB496315 were inoculated in 5 ml LB medium containing ampicillin.</p>
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| - | </div>
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| - | </fieldset>
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| - |
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| - | <!-- Sequencing -->
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| - | <fieldset class="experiment sequencing">
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| - | <legend>Sequencing</legend>
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| - | <div class="investigator">
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| - | <span class="inv">Investigator:</span>
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| - | <span class="inv-names">Dominik</span>
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| - | </div>
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| - | <div class="aim">
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| - | <span class="aim">Aim:</span>
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| - | <span class="aim-desc">Complete sequencing of pSB1A3-iBB496315.</span>
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| - | </div>
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| - | <div class="exp-content">
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| - | <p>8 new sequencing samples were sent out.</p>
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| - | </div>
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| - | </fieldset>
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| - |
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| - | <!-- Compis -->
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| - | <fieldset class="experiment competent cells">
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| - | <legend>Competent cells</legend>
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| - | <div class="investigator">
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| - | <span class="inv">Investigator:</span>
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| - | <span class="inv-names">Dominik</span>
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| - | </div>
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| - | <div class="aim">
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| - | <span class="aim">Aim:</span>
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| - | <span class="aim-desc">Overnight culture for making new aliquots of competent cells.</span>
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| - | </div>
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| - | <div class="exp-content">
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| - | <p>For making new aliquots of <i>E. coli</i> DH5α cells 50 ml LB medium were inoculated with 500 µl of an <i>E. coli</i> DH5α culture.</p>
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| - | </div>
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| - | </fieldset>
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| - |
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| - |
| |
| - | <!-- Miniprep -->
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| - | <fieldset class="experiment miniprep">
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| - | <legend><a name="min">Miniprep</a></legend>
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| - | <div class="investigator">
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| - | <span class="inv">Investigator:</span>
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| - | <span class="inv-names">Dominik</span>
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| - | </div>
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| - | <div class="aim">
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| - | <span class="aim">Aim:</span>
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| - | <span class="aim-desc">preparation of the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9.</span>
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| - | </div>
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| - | <div class="exp-content">
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| - | <p>The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.</p>
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| - | </div>
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| - | </fieldset>
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| - |
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| - |
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| - | </div>
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| - |
| |
| - | </html>
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| - |
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| - | {{:Team:Marburg/Template:ContentEnd}}
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| - | {{:Team:Marburg/Template:Footer}}
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