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| − | <h2>Interlab Study</h2> | + | <h2>Our Team</h2> |
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| − | <h3>Quantifying different sources of variation in gene expression<br> using
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| − | fluorescent transformed E. coli bacteria</h3>
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| − | <h5>
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| − | </h5>
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| − | <h4>Summary:</h4>
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| − | <h5></h5>
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| − | <h5>As part of the 2016 iGEM Interlab Study, we tested 3 different
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| − | fluorescent protein expression plasmids, J23101, J23106, and J23117 with
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| − | positive and negative controls. We transformed the parts with iGEM's
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| − | transformation protocol. Our bacteria were cultured on a LB agar plate and
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| − | cultured in LB broth. To measure the differences in fluorescence expressed
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| − | by different strength plasmids, we used a plate reader with an OD600
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| − | reference point and FITC standard curve.</h5>
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| − | <br><br>
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| − | <h4>Project Description:</h4>
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| − | <h5></h5>
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| − | <h5>The 2016 iGEM Interlab Study aims to compare results obtained by various
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| − | teams in order to quantify the expression of 5 different constructs using
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| − | fluorescence, which provides a useful insight into expression levels that
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| − | can be monitored without disrupting cells. Each device used in the study is
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| − | in the pSB1C3 plasmid backbone and are fluorescent protein expression
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| − | plasmids of different strength promoters. The devices we tested were
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| − | J23101, J23106, J23117 (Test Devices 1, 2, and 3). We used positive control
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| − | I20270 and negative control R0040. J23101, with a strong promoter, showed
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| − | the highest amount of fluorescence. J23117, with a weak promoter, showed
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| − | the lowest amount of fluorescence, and J23106, with the medium strength
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| − | promoter, showed fluorescence amounts between the other two. Our test
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| − | devices were inserted into DH5alpha E. coli, which were transformed
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| − | according to the iGEM transformation protocol. Our transformed cells were
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| − | plated on LB plates with chloramphenicol and incubated overnight at 37° C.
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| − | Two colonies were picked from each plate (with the exception of Test Device
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| − | 3, with which there was only one) and were inoculated in 15mL test tubes.
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| − | These devices were diluted, and then measured in a plate reader with an
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| − | excitation wavelength of 35nm, according to iGEM’s measurement
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| − | protocol.</h5>
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| − | <h4>Protocol:</h4>
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| − | <h5></h5>
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| − | <h5><a href=
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| − | "https://static.igem.org/mediawiki/2016/c/c5/InterLab_iGEM2016_Plate_Reader_Protocol_Updated_July.pdf">
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| − | Plate Reader Protocol</a>
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| − | </h5>
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| − | <h5><a href=
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| − | "https://static.igem.org/mediawiki/parts/6/67/IGEM_Registry_-_Transformation_Protocol.pdf">
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| − | Transformation Protocol</a>
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| − | </h5>
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| − | <h5>
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| − | </h5>
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| − | <br><br>
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| − | <h4>Results:</h4>
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| − | <h5></h5>
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| − | <center>
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| − | <center>
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| − | <table class="image">
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| − | <tr>
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| − | <td><img alt="Placeholder Image" src=
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| − | "http://i.imgur.com/FcVrzL0.png" style="width:800px;">
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| − | </td>
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| − | </tr>
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| − | <td class="caption">Graph of Absorbance (600) over a 6 hour
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| − | time period</td>
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| − | </table>
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| − | </center>
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| − | <table class="image">
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| − | <td><img alt="Placeholder Image" src=
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| − | "http://i.imgur.com/eKgjQDe.png" style="width:800px;">
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| − | </td>
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| − | <td class="caption">Graph of Fluorescence in AU over a 6
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| − | hour time period</td>
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| − | </tr>
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| − | </table>
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| − | </center>
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| − | <table class="image">
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| − | <tr>
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| − | <td><img alt="Placeholder Image" src=
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| − | "http://i.imgur.com/oyXnJRE.png" style="width:800px;">
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| − | </td>
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| − | </tr>
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| − | <td class="caption">Graph of Fluorescence/Absorbance in AU
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| − | over a 6 hour time period</td>
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| − | </tr>
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| − | </table>
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| − | </center>
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| − | <table class="image">
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| − | <tr>
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| − | <td><img alt="Placeholder Image" src=
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| − | "http://i.imgur.com/lWwA42t.png" style="width:800px;">
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| − | </td>
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| − | </tr>
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| − | <td class="caption">FITC Fluorescence standard curve. Row A had
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| − | pipetting errors and so is different from the other
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| − | curves.</td>
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| − | </tr>
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| − | </table>
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| | <script> | | <script> |
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