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| | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Chassis and Safety</h3> | | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px; margin-top:0;">Chassis and Safety</h3> |
| | <h4><strong> What chassis did you use?</strong> </h4> | | <h4><strong> What chassis did you use?</strong> </h4> |
| − | <p>Escherichia coli DH5alpha</p> | + | <p>Escherichia coli DH5α</p> |
| | <h4><strong> What Biosafety Level is your chassis? </strong> </h4> | | <h4><strong> What Biosafety Level is your chassis? </strong> </h4> |
| | <p>BSL1</p> | | <p>BSL1</p> |
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| | <li class="dropdown" style="list-style-type:none;"> | | <li class="dropdown" style="list-style-type:none;"> |
| | <button href="#" class="dropdown-toggle" data-toggle="dropdown"> | | <button href="#" class="dropdown-toggle" data-toggle="dropdown"> |
| − | More details <b class="caret"></b>
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| − | </button>
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| − | <ul class="dropdown-menu" style="padding: 10px; background:#ccc;">
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| − |
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| − | <li>① Set your instrument to read Abs600 (as OD calibration setting)</li>
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| − | <li>② Measure Abs600 of the overnight cultures</li>
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| − | <li>③ Dilute the cultures to a target Abs600 of 0.02 (see the volume of preloading culture and media in Excel Dilution Calculation sheets) in 12 ml LB medium + Chloramphenicol in 50 mL falcon tube.</li>
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| − | <li>④ Remove 500uL of each culture for your zero time point and hold these samples on ice. (You should have 16 sample tubes from the time=0 time point)</li>
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| − | <li>⑤ Incubate the cultures at 37°C and 220 rpm.</li>
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| − | <li>⑥ Remove 500uL samples of each culture after 2, 4 and 6 hours of growth. Keep samples on ice while completing the additional time points. You should have 16 sample tubes for each time point.</li>
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| − | <li>⑦ Set up your four measurement plates: For colony #1 culture, pipet 100 uL samples into wells A, B, C and D of the row for that device. For colony #2, pipet 100uL samples into wells E, F, G, and H of the row for that device.</li>
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| − | <li>⑧ Read your plates, taking care to use the exact same settings used for your fluorescein measurement.</li>
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| − | </ul>
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| − | </li>
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| − | <h5><strong>The initial OD600 measurement of our overnight cultures.</strong> </h5>
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/1/1a/2017_HUST_China_interlab_img_table1.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
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| − | </div>
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| − | <div class="col-xs-12">
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| − | <h5><strong>What type of media did you use for this step? </strong> </h5>
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| − | <p>Luria Bertani </p>
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| − | <h5><strong>What type of vessel or container did you use to grow your cells? </strong> </h5>
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| − | <p>50 ml Falcon tube</p>
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| − | <h5><strong>What temperature setting did you use during the measurement?</strong> </h5>
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| − | <p>22℃</p>
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| − | <h5><strong>What type of 96-well plates did you use?</strong> </h5>
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| − | <p></p>
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| − | <h5><strong>Black plates with transparent/clear bottom (preferred)</strong> </h5>
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| − | <p>Flat</p>
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| − | <h5><strong>Measurement </strong> </h5>
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| − | <ul class="yuandian">
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| − | <li>Measure OD and fluorescence of all samples</li>
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| − | </ul>
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| − | <h5><strong>Suggested Plate Layout for 96-well Plate</strong> </h5>
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| − | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/>
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| − | <a href="https://static.igem.org/mediawiki/2017/8/87/2017_HUST_China_interlab_img_003.png" rel="lightbox-demo" title="my caption"><br />
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/8/87/2017_HUST_China_interlab_img_003.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
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| − | </a>
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| − | </div>
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| − | </div>
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| − | <div id="section7" style="border: solid 1px #666; margin:5px; padding:5px;border-radius:10px;overflow: hidden;">
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| − | <h3 style="background-color:#ce6b9a;color:#ffffff; padding:10px;letter-spacing:1px;margin-top:0;">Interlab Results</h3>
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| − | <h5 class="col-xs-12"><strong>OD600 Reference Point</strong> </h5>
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/5/52/2017_HUST_China_interlab_img_table2.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
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| − | <h5 class="col-xs-12"><strong>Fluorescence standard curve</strong> </h5>
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| − | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/>
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| − | <a href="https://static.igem.org/mediawiki/2017/a/a1/2017_HUST_China_interlab_img_table3.png" rel="lightbox-demo" title="my caption"><br />
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/a/a1/2017_HUST_China_interlab_img_table3.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
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| − | </a>
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| − | <p class="col-xs-12"><strong>NOTE: 50uM Sample exceeds the range of measurements</strong> </p>
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| − | <h5 class="col-xs-12"><strong>OD600 Reference Point</strong> </h5>
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/d/d1/2017_HUST_China_interlab_img_table4.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/d/df/2017_HUST_China_interlab_img_table5.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
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| − | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/>
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| − | <a href="https://static.igem.org/mediawiki/2017/b/b3/2017_HUST_China_interlab_img_table6.png" rel="lightbox-demo" title="my caption"><br />
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/b/b3/2017_HUST_China_interlab_img_table6.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
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| − | </a>
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| − | <p class="col-xs-12"><strong>Final scaling level determined from medium-high points likely to be less impacted by saturation or pipetting error.</strong> </p>
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| − | <p class="col-xs-12"><strong>If needed, you can shift which points are used, but it is likely better to correct instrument settings and protocol.</strong> </p>
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| − | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/>
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| − | <a href="https://static.igem.org/mediawiki/2017/b/b8/2017_HUST_China_interlab_img_table7.png" rel="lightbox-demo" title="my caption"><br />
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/b/b8/2017_HUST_China_interlab_img_table7.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
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| − | </a>
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| − | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/>
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| − | <a href="https://static.igem.org/mediawiki/2017/e/ea/2017_HUST_China_interlab_img_table8.png" title="my caption"><br />
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/e/ea/2017_HUST_China_interlab_img_table8.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
| |
| − | </a>
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| − | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/>
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| − | <a href="https://static.igem.org/mediawiki/2017/d/d2/2017_HUST_China_interlab_img_table9.png" rel="lightbox-demo" title="my caption"><br />
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/d/d2/2017_HUST_China_interlab_img_table9.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
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| − | </a>
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| − | <p><strong><span class="col-xs-12">Click on the picture and you can see a clearer picture.Then click on the return key of the browser and you can return.</span></strong></p><br/>
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| − | <a href="https://static.igem.org/mediawiki/2017/2/2b/2017_HUST_China_interlab_img_table10.png" rel="lightbox-demo" title="my caption" style="padding: 10px;"><br />
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| − | <img title="demo1" src="https://static.igem.org/mediawiki/2017/2/2b/2017_HUST_China_interlab_img_table10.png" alt="demo1" class="col-xs-8 col-xs-offset-2">
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| − | <h3 class="col-xs-12"><strong>Acknowledgments:</strong></h3>
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