Team:SCU-China/Transmitter
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- | + | <li>The Notebook of Transmitter Group</p><p>Since our team is sorted into 4 groups, our team is focusing on the construction of the Transmitter Cell. The whole work that we should do is to build 2 gene lines which lie in the transmitter cells.</p><p>In detail, we use 9 biobricks supplied by IGEM Competition totally which are RBS (BBa_B0030), DT (BBa_B0015), pLux (BBa_R0062), LuxR (BBa_C0062), LasI (BBa_K081016 and BBa_K081009), CinR (BBa_K0077), RhlR (BBa_C0171), and RhlI (BBa_C0051).</p><p>Week 1 7.20-7.26</p><p>1. We transformed the plasmids (BBa_B0030, BBa_B0015, BBa_R0062, BBa_K081016, BBa_K081009, BBa_K0077, BBa_C0171, BBa_C0051) into DH5α E.Coli to amplify these plasmid</p><p>We extracted the plasmids (mentioned above) and used electrophoresis to test the concentration and quality of our plasmids.</p><p>Results:</p><table class="table table-striped"><thead><tr><td><p>Name</li> | |
- | <li>Notebook of | + | |
- | + | ||
- | + | ||
</td><td><p>Concentration (ng/3 μl)</p> | </td><td><p>Concentration (ng/3 μl)</p> | ||
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr></thead><tbody><tr><td><p>BBa_R0062</p> |
- | </td><td><p> | + | </td><td><p>53.3/ 43.5</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_K081016</p> |
- | </td><td><p> | + | </td><td><p>107.9/ 176.7</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_B0030</p> |
- | </td><td><p> | + | </td><td><p>28.2,/8.96</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_C0171</p> |
- | </td><td><p> | + | </td><td><p>322.6/ 387.5</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_K0077</p> |
- | </td><td><p> | + | </td><td><p>193.4,/143.9</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_K081009</p> |
- | </td><td><p> | + | </td><td><p>64.4,/63.1</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_C0077</p> |
- | </td><td><p> | + | </td><td><p>152.7/ 156</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_C0062</p> |
- | </td><td><p> | + | </td><td><p>53.29/128.7</p> |
</td> | </td> | ||
</tr><tr><td><p>BBa_B0015</p> | </tr><tr><td><p>BBa_B0015</p> | ||
- | </td><td><p> | + | </td><td><p>116.2/19.5</p> |
</td> | </td> | ||
- | </tr> | + | </tr></tbody> |
- | </ | + | </table> |
- | < | + | <li>Note: The different concentrations are from different tubes.</p><p>Week 2 7.27-8.03</p><p>1. We transformed the BBa_C0062 and BBa_C0077 plasmids.</p><p>2. We extracted the BBa_J37019, BBa_I9026, BBa_K081005, BBa_I714075, BBa_K93600 and BBa_R0077 plasmids and determined the concentration of them by electrophoresis.</p><p>3. We cleaved the BBa_R0062, BBa_C0051, BBa_K0077 plasmids with EcoR I and Spe I enzyme and BBa_B0015 with Xba I and Pst I. Also, we prepared the pSB1A3 and pSB1K3 backbones with EcoR I and Pst I.</p><p>4. We cleaved the BBa_J37019 (Named E11) and BBa_R0077 (E21) with EcoR I and Spe I, BBa_I9026 (E12), BBa_K081016 (E22) and BBa_C0077 (E24) with Xba I and Pst I.</p><p>5. We linked the E11 and E12 with pSB1A3 (L11), the E21 and E22 with pSB1A3 (L21).</p><p>6. We transformed all the linked parts into E.coli.</p><p>7. We cleaved BBa_B0015 with Xba I and Pst I.</p><p>Results</p><table class="table table-striped"><caption>The concentration of plasmids</caption><thead><tr><td><p>Name</li> |
- | <li>Note: | + | |
- | + | </td><td><p>Concentration (ng/3μl)</p> | |
- | + | ||
- | </td><td><p>Concentration (ng/3 μl)</p> | + | |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr></thead><tbody><tr><td><p>BBa_J37019</p> |
- | </td><td><p> | + | </td><td><p>400/403</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_I9026</p> |
- | </td><td><p> | + | </td><td><p>247/140.6</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_R0077</p> |
- | </td><td><p> | + | </td><td><p>134/161.2</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_K081005</p> |
- | </td><td><p> | + | </td><td><p>51.2/27.6</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_I714075</p> |
- | </td><td><p> | + | </td><td><p>95.7/37.2</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_K93600</p> |
- | </td><td><p> | + | </td><td><p>41.6</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>L1</p> |
- | </td><td><p> | + | </td><td><p>92.3/54.2</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>L2</p> |
- | </td><td><p> | + | </td><td><p>88.4/60.4</p> |
</td> | </td> | ||
- | </tr>< | + | </tr></tbody> |
- | </ | + | </table> |
+ | <li>Note: The different concentrations are from different tubes.</p><p>2. The electrophoresis results are shown in Figure 1.</p><p>Figure 1. The cleaved plasmids</p><p>Week 3 8.04-8.10</p><p>1. We linked the L1 and BBa_B0015 with pSB1T3 (named F1), L2 and BBa_I9026 with pSB1T3 (F2). Then we transformed them into E.coli to amplify them and we extracted the plasmids to test their quality.</p><p>2. We linked E11 and E12 with pSB1T3 (named LA1), E21 and E22 with pSB1A3 (LA2).</p><p>3. We cleaved the BBa_J37019 (Named E11) and BBa_R0077 (E21) with EcoR I and Spe I, BBa_I9026 (E12), BBa_K081016 (E22) and BBa_C0077 (E24) with Xba I and Pst I again and tested their qualities by electrophoresis and then we did the gel purification.</p><p>4. We cleaved the BBa_R0077 with Spe I and Pst I (named EP21).</p><p>5. We linked EP21 and E22 with 3 different backbones (pSB1T3, pSB1A3, and pSB1C3) which are tested by electrophoresis.</p><p>Results:</p><table class="table table-striped"><caption>The concentration of plasmids and linkage product</caption><thead><tr><td><p>Name</li> | ||
+ | |||
+ | </td><td><p>Concentration (ng/3μl)</p> | ||
</td> | </td> | ||
- | </tr> | + | </tr></thead><tbody><tr><td><p>F11	</p> |
- | </ | + | </td><td><p>Failed</p> |
- | < | + | </td> |
- | <li>Note: | + | </tr><tr><td><p>F21</p> |
- | + | </td><td><p>failed</p> | |
- | + | </td> | |
- | </td><td><p>Concentration (ng/3 μl)</p> | + | </tr><tr><td><p>E11</p> |
+ | </td><td><p>8.53</p> | ||
+ | </td> | ||
+ | </tr><tr><td><p>E12</p> | ||
+ | </td><td><p>Failed</p> | ||
+ | </td> | ||
+ | </tr><tr><td><p>E21</p> | ||
+ | </td><td><p>24.9/7.5</p> | ||
+ | </td> | ||
+ | </tr><tr><td><p>E22</p> | ||
+ | </td><td><p>14/5.3</p> | ||
+ | </td> | ||
+ | </tr><tr><td><p>E24</p> | ||
+ | </td><td><p>18.1/17.7</p> | ||
+ | </td> | ||
+ | </tr></tbody> | ||
+ | </table> | ||
+ | <li>Note: The different concentrations are from different tubes.</p><p>2. <img width="60" height="29" src="Notebook of Transmitter(1).files/Notebook of Transmitter(1)3160.png"><img width="79" height="29" src="Notebook of Transmitter(1).files/Notebook of Transmitter(1)3161.png">The electrophoresis results are shown in Figure 2.</p><p>Figure 2. A, C, and D the gel purification results and cleaved plasmids. B. the cleaved EP21.</p><p>Week 4 8.11-8.17</p><p>1. We did the liquid culture of BBa_R0077 and BBa_C0077.</p><p>2. We extracted the BBa_J37019, BBa_I9026 plasmids and then we cleaved both of them (BBa_J37019 with Spe I and Pst I; BBa_I9026 with Xba I and Pst I) and linked them with pSB1C3. Finally we tested the quality by transforming into E.coli.</p><p>3. We linked BBa_J37019 and BBa_I9026(LI1), BBa_R0077 and BBa_C0077 with pSB1C3.</p><p>4. We extracted BBa_I9026, BBa_J37019, BBa_C0077, and BBa_R0077.</p><p>5. We cleaved BBa_J37019 with E, S (D1, D2) or S, P (D3, D4) and BBa_I9026 plasmids with E, S, (D5, D6) and S, P (D7, D8) enzymes. Then we did the gel purification.</p><p>6. We linked the BBa_J37019 with BBa_I9026 with different types (named LI3).</p><p>7. We transformed LI3 into E.coli and extracted the plasmids to test the quality.</p><p>Results</p><table class="table table-striped"><caption>The concentration of plasmids</caption><thead><tr><td><p>Name</li> | ||
+ | |||
+ | </td><td><p>Concentration (ng/3μl)</p> | ||
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr></thead><tbody><tr><td><p>BBa_I9026</p> |
- | </td><td><p> | + | </td><td><p>289.2/112.6</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_J37019</p> |
- | </td><td><p> | + | </td><td><p>193.2/40.7</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_C0077</p> |
- | </td><td><p> | + | </td><td><p>27.7/22.5/33.3</p> |
</td> | </td> | ||
- | </tr><tr><td><p> | + | </tr><tr><td><p>BBa_R0077</p> |
- | </td><td><p> | + | </td><td><p>177.8/179.8/200.3</p> |
</td> | </td> | ||
- | </tr> | + | </tr></tbody> |
- | </ | + | </table> |
- | < | + | <p>Note: The different concentrations are from different tubes.</p><p>2. The electrophoresis results are shown in Figure 3.</p><p>Figure A. the cleaved BBa_J37019 and BBa_I9026 plasmids. B. the cleaved linkage products. C and D is the gel purification of BBa_J37019 and BBa_I9026. E. the cleaved BBa_R0077 and BBa_C0077.</p><p>F. the linkage products.</p><p>Week 5 8.18-8.24</p><p>1. We linked the E11 and E12 with pSB1T3 by 3A assembly and E11 and E12 by traditional assembly.</p><p>2. We Amplified the LII3 which was tested by cleavage and electrophoresis.</p><p>3. We transformed the BBa_J37019, BBa_I9026, BBa_C0077, and L1T1.</p><p>4. We cultured the LIIF in liquid and transformed it.</p><p>5. We did the gel purification of BBa_J37019 and BBa_K747092.</p><p>Results</p><p>The electrophoresis results are shown in Figure 4.</p><p>Figure 4. A the gel purification results; B. the LII3 results.</li> |
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Revision as of 03:34, 18 October 2014