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| | {{:Team:Marburg/Template:Header}} | | {{:Team:Marburg/Template:Header}} |
| | {{:Team:Marburg/Template:ContentTitleNav}} | | {{:Team:Marburg/Template:ContentTitleNav}} |
| - | Notebook: August | + | Notebook: August <html><a href="https://2013.igem.org/Team:Marburg/Notebook:September"><img src="https://static.igem.org/mediawiki/2013/7/71/Mr-igem-next-arrow.png" style="float:right;margin-left:5px !important;" alt="Next"></a> <a href="https://2013.igem.org/Team:Marburg/Notebook:July"><img src="https://static.igem.org/mediawiki/2013/1/13/Mr-igem-previous-arrow.png" alt="Previous" style="float:right;"></a></html> |
| - | {{:Team:Marburg/Template:ContentStartNav}}
| + | {{:Team:Marburg/Template:ContentStartNav}}<html> |
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| - | <html>
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| - | <div class="notebooky-entry"> | + | |
| - | <h2 class="title">
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| - | <a name=" 01-08-2013"> 01. 08.2013 </ a> | + | |
| - | </h2>
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| - | | + | |
| - | <fieldset class="experiment sequencing">
| + | |
| - | <legend>Sequencing</le{{:Team:Marburg/Template:Header}}
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| - | {{:Team:Marburg/Template:ContentTitleNav}}
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| - | Notebook: August
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| - | {{:Team:Marburg/Template:ContentStartNav}} | + | |
| - | | + | |
| - | <html> | + | |
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| | <div class="notebooky-entry"> | | <div class="notebooky-entry"> |
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| | <li>0.3 µl <i>Pst</I></li> | | <li>0.3 µl <i>Pst</I></li> |
| | <li>1 µl CutSmart</li> | | <li>1 µl CutSmart</li> |
| - | <li>7.4 µl ddH2O</li> | + | <li>7.4 µl ddbidest. H2O</li> |
| | </ul> | | </ul> |
| - | <p>The samples were incubated for 1 h at 37° C.</p> | + | <p>The samples were incubated for 1 h at 37 °C.</p> |
| | <p>After that, a gel electrophoresis was made, but failed and has to be repeated the next day</p> | | <p>After that, a gel electrophoresis was made, but failed and has to be repeated the next day</p> |
| | </div> | | </div> |
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| | <div class="exp-content"> | | <div class="exp-content"> |
| | <p>The chemo competent <i>E. coli</i> DH5α cells were transformed with the plasmids pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315 and plated on LB-Amp-plates.<br /> | | <p>The chemo competent <i>E. coli</i> DH5α cells were transformed with the plasmids pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315 and plated on LB-Amp-plates.<br /> |
| - | The plates were incubated over night at 37° C.</p> | + | The plates were incubated over night at 37 °C.</p> |
| | </div> | | </div> |
| | </fieldset> | | </fieldset> |
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| | <li>0.3 µl <i>PstI</i></li> | | <li>0.3 µl <i>PstI</i></li> |
| | <li>1 µl CutSmart</li> | | <li>1 µl CutSmart</li> |
| - | <li>7.4 µl ddH2O</li> | + | <li>7.4 µl ddbidest. H2O</li> |
| | </ul> | | </ul> |
| - | <p>The samples were incubated for 1 h at 37° C.</p> | + | <p>The samples were incubated for 1 h at 37 °C.</p> |
| | <table class="gel digest"> | | <table class="gel digest"> |
| | <colgroup> | | <colgroup> |
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| | </ul> | | </ul> |
| | <p>All plasmids are brought to Marian who will transform them into <i>P. tricornutum</i> cells.</p> | | <p>All plasmids are brought to Marian who will transform them into <i>P. tricornutum</i> cells.</p> |
| - | </div>
| |
| - | </fieldset>
| |
| - | </div>
| |
| - |
| |
| - | </html>
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| - |
| |
| - | {{:Team:Marburg/Template:ContentEndNav}}
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| - | {{:Team:Marburg/Template:Footer}}
| |
| - | gend>
| |
| - | <div class="investigator">
| |
| - | <span class="inv">Investigator:</span>
| |
| - | <span class="inv-names">Dominik</span>
| |
| - | </div>
| |
| - | <div class="aim">
| |
| - | <span class="aim">Aim:</span>
| |
| - | <span class="aim-desc">Examination of the sequence of pSB1A3-iBB4+iBB10+iBB96315.</span>
| |
| - | </div>
| |
| - | <div class="exp-content">
| |
| - | <p>2 additional sequence samples were sent out for sequencing.</p>
| |
| - | </div>
| |
| - | </fieldset>
| |
| - |
| |
| - | <fieldset class="experiment digest">
| |
| - | <legend><a name="dig">Digest</a></legend>
| |
| - | <div class="investigator">
| |
| - | <span class="inv">Investigator:</span>
| |
| - | <span class="inv-names">Dominik</span>
| |
| - | </div>
| |
| - | <div class="aim">
| |
| - | <span class="aim">Aim:</span>
| |
| - | <span class="aim-desc">Test digest of pSB1A3-iBB4+iBB13+iBB96315 with <i>Eco</i>RI and <i>Pst</i>I</span>
| |
| - | </div>
| |
| - | <div class="exp-content">
| |
| - | <p><ul class="digest">
| |
| - | <li>1 µl Plasmid (pSB1C3-iBB39,pSB1A3-iBB49 and pSB1C3-iBB6315)</li>
| |
| - | <li>0.3 µl <i>Eco</i>RI</li>
| |
| - | <li>0.3 µl <i>Pst</I></li>
| |
| - | <li>1 µl CutSmart</li>
| |
| - | <li>7.4 µl ddH2O</li>
| |
| - | </ul>
| |
| - | <p>The samples were incubated for 1 h at 37° C.</p>
| |
| - | <p>After that, a gel electrophoresis was made, but failed and has to be repeated the next day</p>
| |
| - | </div>
| |
| - | </fieldset>
| |
| - |
| |
| - | <fieldset class="experiment inoculation">
| |
| - | <legend><a name="ino">Inoculation</a></legend>
| |
| - | <div class="investigator">
| |
| - | <span class="inv">Investigator:</span>
| |
| - | <span class="inv-names">Dominik</span>
| |
| - | </div>
| |
| - | <div class="aim">
| |
| - | <span class="aim">Aim:</span>
| |
| - | <span class="aim-desc">inoculation of colonies for plasmid preparation</span>
| |
| - | </div>
| |
| - | <div class="exp-content">
| |
| - | <p>4 colonies of pSB1A3-iBB496315 were inoculated in 5 ml LB medium containing ampicillin.</p>
| |
| - | </div>
| |
| - | </fieldset>
| |
| - | </div>
| |
| - |
| |
| - | <div class="notebooky-entry">
| |
| - | <h2 class="title">
| |
| - | <a name="02-08-2013">02.08.2013</a>
| |
| - | </h2>
| |
| - |
| |
| - | <fieldset class="experiment sequencing">
| |
| - | <legend>Sequencing</legend>
| |
| - | <div class="investigator">
| |
| - | <span class="inv">Investigator:</span>
| |
| - | <span class="inv-names">Dominik</span>
| |
| - | </div>
| |
| - | <div class="aim">
| |
| - | <span class="aim">Aim:</span>
| |
| - | <span class="aim-desc">Analysis of the sequence of pSB1A3-iBB496315.</span>
| |
| - | </div>
| |
| - | <div class="exp-content">
| |
| - | <p>Obviously the wrong plasmid was sent out for sequencing. Due to the lack of enough plasmids for an additional sequence analysis, we will redo a new miniprep.</p>
| |
| - | </div>
| |
| - | </fieldset>
| |
| - |
| |
| - | <fieldset class="experiment transformation">
| |
| - | <legend>Transformation</legend>
| |
| - | <div class="investigator">
| |
| - | <span class="inv">Investigator:</span>
| |
| - | <span class="inv-names">Dominik</span>
| |
| - | </div>
| |
| - | <div class="aim">
| |
| - | <span class="aim">Aim:</span>
| |
| - | <span class="aim-desc">Transformation of <i>E. coli</i> DH5α with the plasmid DNA pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.</span>
| |
| - | </div>
| |
| - | <div class="exp-content">
| |
| - | <p>The chemo competent <i>E. coli</i> DH5α cells were transformed with the plasmids pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315 and plated on LB-Amp-plates.<br />
| |
| - | The plates were incubated over night at 37° C.</p>
| |
| - | </div>
| |
| - | </fieldset>
| |
| - | </div>
| |
| - |
| |
| - | <div class="notebooky-entry">
| |
| - | <h2 class="title">
| |
| - | <a name="03-08-2013">03.08.2013</a>
| |
| - | </h2>
| |
| - |
| |
| - | <fieldset class="experiment miniprep">
| |
| - | <legend><a name="min">Miniprep</a></legend>
| |
| - | <div class="investigator">
| |
| - | <span class="inv">Investigator:</span>
| |
| - | <span class="inv-names">Dominik</span>
| |
| - | </div>
| |
| - | <div class="aim">
| |
| - | <span class="aim">Aim:</span>
| |
| - | <span class="aim-desc">preparation of the plasmid pSB1A3-iBB496315.</span>
| |
| - | </div>
| |
| - | <div class="exp-content">
| |
| - | <p>The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.</p>
| |
| - | </div>
| |
| - | </fieldset>
| |
| - |
| |
| - | <fieldset class="experiment digest">
| |
| - | <legend><a name="dig">Digest</a></legend>
| |
| - | <div class="investigator">
| |
| - | <span class="inv">Investigator:</span>
| |
| - | <span class="inv-names">Dominik</span>
| |
| - | </div>
| |
| - | <div class="aim">
| |
| - | <span class="aim">Aim:</span>
| |
| - | <span class="aim-desc">Test digest of pSB1A3-iBB496315 and iBB4+iBB13+iBB96315 with <i>Eco</i>RI and <i>Pst</i>I</span>
| |
| - | </div>
| |
| - | <div class="exp-content">
| |
| - | <p><ul class="digest">
| |
| - | <li>1 µl Plasmid (pSB1A3-iBB496315 and iBB4+iBB13+iBB96315)</li>
| |
| - | <li>0.3 µl EcoRI</li>
| |
| - | <li>0.3 µl PstI</li>
| |
| - | <li>1 µl CutSmart</li>
| |
| - | <li>7.4 µl ddH2O</li>
| |
| - | </ul>
| |
| - | <p>The samples were incubated for 1 h at 37° C.</p>
| |
| - | <table class="gel digest">
| |
| - | <colgroup>
| |
| - | <col width="50%" />
| |
| - | <col width="50%" />
| |
| - | </colgroup>
| |
| - | <thead>
| |
| - | <tr>
| |
| - | <th colspan="2" class="title">Gel electrophoresis</th>
| |
| - | </tr>
| |
| - | </thead>
| |
| - | <tbody>
| |
| - | <tr>
| |
| - | <td>
| |
| - | <img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
| |
| - | </td>
| |
| - | <td>
| |
| - | <p>
| |
| - | <span class="gel-elc">Gel substances</span>
| |
| - | <ul class="gel-sub">
| |
| - | <li>1% Agarose gel</li>
| |
| - | <li>10 µl RedSafe in 50 ml gel</li>
| |
| - | <li>x µl Hyper Ladder</li>
| |
| - | </ul>
| |
| - | </p>
| |
| - | <p>
| |
| - | <span class="exp">Expectations</span>
| |
| - | <ul class="exp">
| |
| - | <li>Lane 1: 5300 kbp</li>
| |
| - | <li>Lane 2: 3300 kbp</li>
| |
| - | <li>Lane 3: 1300 kbp</li>
| |
| - | </ul>
| |
| - | </p>
| |
| - | <p>Worked as expected.</p>
| |
| - | </td>
| |
| - | </tr>
| |
| - | </tbody>
| |
| - | </table>
| |
| - | </div>
| |
| - | </fieldset>
| |
| - | </div>
| |
| - |
| |
| - | <div class="notebooky-entry">
| |
| - | <h2 class="title">
| |
| - | <a name="14-08-2013">14.08.2013</a>
| |
| - | </h2>
| |
| - |
| |
| - | <fieldset class="experiment sequencing">
| |
| - | <legend><a name="seq">Sequencing</a></legend>
| |
| - | <div class="investigator">
| |
| - | <span class="inv">Investigator:</span>
| |
| - | <span class="inv-names">Dominik</span>
| |
| - | </div>
| |
| - | <div class="aim">
| |
| - | <span class="aim">Aim:</span>
| |
| - | <span class="aim-desc">Examination of sequence analysis of the plasmids pSB1A3-iBB496315, pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.</span>
| |
| - | </div>
| |
| - | <div class="exp-content">
| |
| - | <p><ul class="seq">
| |
| - | <li>pSB1A3-iBB496315: Okay</li>
| |
| - | <li>pSB1A3-iBB4+iBB10+iBB96315: mutation in signal peptide that leads to Trp → Leu. Will be ignored</li>
| |
| - | <li>pSB1A3-iBB4+iBB13+iBB96315: mutation in terminator, will be ignored.</li>
| |
| - | </ul>
| |
| - | <p>All plasmids are brought to Marian who will transform them in <i>P. tricornutum</i> cells.</p>
| |
| | </div> | | </div> |
| | </fieldset> | | </fieldset> |
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