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<div class="main"> | <div class="main"> | ||
− | <a href="#" | + | <a href="#" Onclick="show(0)"> |
− | + | March 15th- April 13th | |
</a> | </a> | ||
<div id="child1" style="display:none;"> | <div id="child1" style="display:none;"> | ||
<p>We noticed Igem competition and get acquainted with synthesis biology. </p> | <p>We noticed Igem competition and get acquainted with synthesis biology. </p> | ||
</div> | </div> | ||
− | <br> | + | <br> <a href="#" OnClick="show(1)"> |
April 14th-May 1st | April 14th-May 1st | ||
</a> | </a> | ||
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<p>Our team, NWU-CHINA, had been organized. Then we find our PI and confirmed our project. Our PI gave us a lot of advice about following experiment.</p> | <p>Our team, NWU-CHINA, had been organized. Then we find our PI and confirmed our project. Our PI gave us a lot of advice about following experiment.</p> | ||
</div> | </div> | ||
− | <br> <a href="#" | + | <br> <a href="#" OnClick="show(2)"> |
May 3rd-May 20th | May 3rd-May 20th | ||
</a> | </a> | ||
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<p>We accepted foundational lab training and learned necessary experiment operation. </p> | <p>We accepted foundational lab training and learned necessary experiment operation. </p> | ||
</div> | </div> | ||
− | <br> <a href="#" | + | <br> <a href="#" OnClick="show(3)"> |
June 1st-June 20th | June 1st-June 20th | ||
</a> | </a> | ||
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<p>We cleared Interlab Study requirement, RFC and 3A assembly protocol. Meanwhile, we found a lot of reference paper to find theory supporting our experiment. </p> | <p>We cleared Interlab Study requirement, RFC and 3A assembly protocol. Meanwhile, we found a lot of reference paper to find theory supporting our experiment. </p> | ||
</div> | </div> | ||
− | <br> <a href="#" | + | <br> <a href="#" OnClick="show(4)"> |
July 15th-July 26th | July 15th-July 26th | ||
</a> | </a> | ||
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<p>We finished our First Interlab Study</p> | <p>We finished our First Interlab Study</p> | ||
</div> | </div> | ||
− | <br> <a href="#" | + | <br> <a href="#" OnClick="show(5)"> |
August 1st-August 31st | August 1st-August 31st | ||
</a> | </a> | ||
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<li>gene by agarose gel electrophoresis and purified the gene by gel slices. </li> | <li>gene by agarose gel electrophoresis and purified the gene by gel slices. </li> | ||
<li>We used pAK1900 as our vector. Then we used BamH I and Sac I to digest pAK1900 and purified gene for 5 hours.</li> | <li>We used pAK1900 as our vector. Then we used BamH I and Sac I to digest pAK1900 and purified gene for 5 hours.</li> | ||
− | <li>Then we used T4 DNA Ligase to ligated our vector and RFP gene at 4 | + | <li>Then we used T4 DNA Ligase to ligated our vector and RFP gene at 4 ° for 24 hours</li> |
<li>We transferred our vector into E.coli DH5&alpha competent cell to get more vector clones. Our transferring protocol is as follows:</li> | <li>We transferred our vector into E.coli DH5&alpha competent cell to get more vector clones. Our transferring protocol is as follows:</li> | ||
<ol type="a"> | <ol type="a"> | ||
− | <li>Mix 4ul vector and 50ul competent cells in 1.5mL centrifuge tube at 0 | + | <li>Mix 4ul vector and 50ul competent cells in 1.5mL centrifuge tube at 0 ° for 30 min. </li> |
− | <li>Hot shock for 90 seconds at | + | <li>Hot shock for 90 seconds at 42° . </li> |
− | <li>Keep competent cell at | + | <li>Keep competent cell at 0° for 4-5 min, then add 950µL Soc media into the tube. </li> |
− | <li>Incubate at 37 | + | <li>Incubate at 37 ° for 2 hours shaking at 220rpm</li> |
− | <li>Centrifuge at 6000rpm for 1min. After centrifugation, 900µL supernatant is discarded, then resuspended cells, spread the culture on plate with antibiotic 100µg/µL TC , incubate at | + | <li>Centrifuge at 6000rpm for 1min. After centrifugation, 900µL supernatant is discarded, then resuspended cells, spread the culture on plate with antibiotic 100µg/µL TC , incubate at 37° for 16 hours. </li> |
− | <li>Select several colonies, purified them on another selective plate, incubate at | + | <li>Select several colonies, purified them on another selective plate, incubate at 37° for 12 hours. </li> |
</ol> | </ol> | ||
<li>Incubate purified cultures with 5mL LB media and 5ul 5mg/µL TC for 5 hours. </li> | <li>Incubate purified cultures with 5mL LB media and 5ul 5mg/µL TC for 5 hours. </li> | ||
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</p> | </p> | ||
</div> | </div> | ||
− | <br> <a href="#" | + | <br> <a href="#" OnClick="show(6)"> |
September 1st-10th | September 1st-10th | ||
</a> | </a> | ||
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<p>We had done our second Interlab Study. Meanwhile, we used PCR to clone GntR and AlkB2 gene. And ligated them on constructed RFP vector. Then we separated cloned gene by agarose gel electrophoresis and purified the gene by gel slices. </p> | <p>We had done our second Interlab Study. Meanwhile, we used PCR to clone GntR and AlkB2 gene. And ligated them on constructed RFP vector. Then we separated cloned gene by agarose gel electrophoresis and purified the gene by gel slices. </p> | ||
</div> | </div> | ||
− | <br> <a href="#" | + | <br> <a href="#" OnClick="show(7)"> |
September 11th-October 12th | September 11th-October 12th | ||
</a> | </a> | ||
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<p><ol> | <p><ol> | ||
<li>We used Hind I, Hind II, Sal I, Hind III and Xba I, Spe I and Sal I digest pAK1900, GntR, AlkB2 for 5 hours respectively. </li> | <li>We used Hind I, Hind II, Sal I, Hind III and Xba I, Spe I and Sal I digest pAK1900, GntR, AlkB2 for 5 hours respectively. </li> | ||
− | <li>Then we used T4 DNA Ligase to ligated them at | + | <li>Then we used T4 DNA Ligase to ligated them at 4° for 24 hours. </li> |
<li>Then we cloned our vector in DH5&alpha competent cells. </li> | <li>Then we cloned our vector in DH5&alpha competent cells. </li> | ||
<li>Sequence our gene to verify our vector. </li> | <li>Sequence our gene to verify our vector. </li> | ||
</ol></p> | </ol></p> | ||
</div> | </div> | ||
− | <br> <a href="#" | + | <br> <a href="#" OnClick="show(8)"> |
October 12th-17th | October 12th-17th | ||
</a> | </a> | ||
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</div> | </div> | ||
</body> | </body> | ||
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Revision as of 07:16, 16 October 2017