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| − | <h2 class="section-heading">Cell measurements </h2>
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| − | <p class="lead">Two colonies from each transformation were picked, and grown in foil-covered 50 ml falcon tubes over night (18 hours). </p>
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| − | <p class="lead"><strong>Preparation</strong> OD was measured, and a dilution was calculated to achieve an OD of 0.02. Dilution calculations can be found in the table next to this. Here we used the calculated correction factor from our initial abs/OD calibration. From the absorption measurements taken at 0 hours, we see indications of pipetting errors, as the OD<sub>600</sub> (average) ranges from 0.015 to 0.6 (table 1). </p>
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| − | <button onclick="javascript:showhide('InterLab-table')">Show table 1</button>
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| − | <div id="InterLab-table" style="display:none;">
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| − | <table class="responsive-table">
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| − | <caption><b>Table 1</b> Starting OD.</caption>
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| − | <thead>
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| − | <tr>
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| − | <th scope="col"></th>
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| − | <th scope="col">Abs<sub>600</sub></th>
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| − | <th scope="col">OD<sub>600</sub> before</th>
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| − | <th scope="col">Volume (ml) added to 12 ml media</th>
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| − | <th scope="col">OD<sub>600</sub> after dilution (0 hours)</th>
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| − | </tr>
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| − | </thead>
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| − | <tfoot>
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| − | </tfoot>
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| − | <tbody>
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| − | <tr>
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| − | <th scope="row">Negative Control (Colony 1)</th>
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| − | <td data-title="Abs600">0,461</td>
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| − | <td data-title="OD600 before">1,435</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,167</td>
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| − | <td data-title="OD600 after dilution (0 hours)" data-type="currency">0,06483974</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Negative Control (Colony 2)</th>
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| − | <td data-title="Abs600">0,463</td>
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| − | <td data-title="OD600 before">1,439</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,167</td>
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| − | <td data-title="OD600 after dilution (0 hours)" data-type="currency">0,02584249</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Positive Control (Colony 1)</th>
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| − | <td data-title="Abs<sub>600</sub>">0,450</td>
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| − | <td data-title="OD<sub>600</sub> before">1,400</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,171</td>
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| − | <td data-title="OD<sub>600</sub> after dilution (0 hours)" data-type="currency">0,04467949</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Positive Control (Colony 2)</th>
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| − | <td data-title="Abs<sub>600</sub>">0,474</td>
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| − | <td data-title="OD<sub>600</sub> before">1,475</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,163</td>
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| − | <td data-title="OD<sub>600</sub> after dilution (0 hours)" data-type="currency">0,01689103</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 1: J23101+I13504 (Colony 1)</th>
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| − | <td data-title="Abs<sub>600</sub>">0,462</td>
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| − | <td data-title="OD<sub>600</sub> before">1,437</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,167</td>
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| − | <td data-title="OD<sub>600</sub> after dilution (0 hours)" data-type="currency">0,03043498</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 1: J23101+I13504 (Colony 2)</th>
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| − | <td data-title="Abs<sub>600</sub>">0,472</td>
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| − | <td data-title="OD<sub>600</sub> before">1,469</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,163</td>
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| − | <td data-title="OD<sub>600</sub> after dilution (0 hours)" data-type="currency">0,01977106</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 2: J23106+I13504 (Colony 1)</th>
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| − | <td data-title="Abs<sub>600</sub>">0,477</td>
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| − | <td data-title="OD<sub>600</sub> before">1,483</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,162</td>
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| − | <td data-title="OD<sub>600</sub> after dilution (0 hours)" data-type="currency">0,03175824</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 2: J23106+I13504 (Colony 2)</th>
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| − | <td data-title="Abs<sub>600</sub>">0,454</td>
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| − | <td data-title="OD<sub>600</sub> before">1,410</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,170</td>
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| − | <td data-title="OD<sub>600</sub> after dilution (0 hours)" data-type="currency">0,018837</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 3: J23117+I13504 (Colony 1)</th>
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| − | <td data-title="Abs<sub>600</sub>">0,507</td>
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| − | <td data-title="OD<sub>600</sub> before">1,577</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,152</td>
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| − | <td data-title="OD<sub>600</sub> after dilution (0 hours)" data-type="currency">0,03261447</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 3: J23117+I13504 (Colony 2)</th>
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| − | <td data-title="Abs<sub>600</sub>">0,492</td>
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| − | <td data-title="OD<sub>600</sub> before">1,529</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,157</td>
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| − | <td data-title="OD<sub>600</sub> after dilution (0 hours)" data-type="currency">0,01471154</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 4: J23101.BCD2.E0040.B0015 (Colony 1)</th>
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| − | <td data-title="Abs<sub>600</sub>">0,420</td>
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| − | <td data-title="OD<sub>600</sub> before">1,307</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,184</td>
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| − | <td data-title="OD<sub>600</sub> after dilution (0 hours)" data-type="currency">0,03759615</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 4: J23101.BCD2.E0040.B0015 (Colony 2)</th>
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| − | <td data-title="Abs600">0,472</td>
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| − | <td data-title="OD600 before">1,468</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,163</td>
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| − | <td data-title="OD600 after dilution (0 hours)" data-type="currency">0,01860348</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 5: J23106.BCD2.E0040.B0015 (Colony 1)</th>
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| − | <td data-title="Abs600">0,513</td>
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| − | <td data-title="OD600 before">1,595</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,150</td>
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| − | <td data-title="OD600 after dilution (0 hours)" data-type="currency">0,04234432</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 5: J23106.BCD2.E0040.B0015 (Colony 2)</th>
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| − | <td data-title="Abs600">0,467</td>
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| − | <td data-title="OD600 before">1,451</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,165</td>
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| − | <td data-title="OD600 after dilution (0 hours)" data-type="currency">0,01494505</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 6: J23117.BCD2.E0040.B0015 (Colony 1)</th>
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| − | <td data-title="Abs600">0,482</td>
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| − | <td data-title="OD600 before">1,498</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,160</td>
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| − | <td data-title="OD600 after dilution (0 hours)" data-type="currency">0,04452381</td>
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| − | </tr>
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| − | <tr>
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| − | <th scope="row">Test Device 6: J23117.BCD2.E0040.B0015 (Colony 2)</th>
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| − | <td data-title="Abs600">0,455</td>
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| − | <td data-title="OD600 before">1,416</td>
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| − | <td data-title="Volume (ml) added to 12 ml media" data-type="currency">0,170</td>
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| − | <td data-title="OD600 after dilution (0 hours)" data-type="currency">0,02008242</td>
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| − | </tr>
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| − | </table>
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| − | <h2 class="section-heading">OD<sub>600</sub </h2>
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| − | <div class="clearfix"></div>
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| − | <br>
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| − | <p class="lead">Cell growth stagnated between 4 and 6 hours. Cells transformed with Test Device 1 and 4 grew slower than the 6 other transformations, and even decreased in OD between 4 and 6 hours. Click on figures for enlarged images:</p>
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| − | <img class="fblogo" border="0" alt="Graph2" src="https://static.igem.org/mediawiki/2017/6/6e/Graph2_InterLab.png"/></a>
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| − | <h2 class="section-heading">Fluorescence</h2>
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| − | <br>
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| − | <p class="lead">Some transformed cells continue to increase fluorescence despite a decrease in OD in the same samples. Devices 3 and 6 are very close to the negative control in fluorescence. Click on figures for enlarged images:</p>
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| − | <img class="fblogo" border="0" alt="Graph3" src="https://static.igem.org/mediawiki/2017/e/e5/Graph3_InterLab.png"/></a>
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| − | <img class="fblogo" border="0" alt="Graph4" src="https://static.igem.org/mediawiki/2017/e/e9/Graph4_InterLab.png"/></a>
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| − | <h2 class="section-heading">Conclusion </h2>
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| − | <p class="lead">Our data indicate which plasmid elements induce the highest production of GFP.
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| − | Plasmids containing J23117 (Test devices 3 and 6) does not express fluorescence to a higher degree than the negative control. Plasmids with J23101 (Device 1 and 4) induced the highest fluorescence, and plasmids containing J23106 (Test devices 2 and 5) were somewhere in between. Combining the J23101 or J23106 with I13504 (Test devices 1-3) gave a higher fluorescence than adding BCD2.E0040.B0015 (Test devices 4-6).
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| − | <br><br>
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| − | These results are not reliable on their own, but will be more robust and reliable when combined with data from the other teams participating in the interlab study.
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