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Team:SCU-China/Sensor
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| - | <p>Since our team are sorted into 4 groups our target is constructing the Sensor cell in our system. According to our design, we need to construct two gene pathways. Consequently, we use the following parts: pLac (BBa_R0080), Promoter C and arabinose C operon (BBa_I13458), RBS (BBa_B0030), LuxI (BBa_C0061), double terminator (BBa_B0015), Constitutive promoter (BBa_J23100), CinI (BBa_C0076), pBAD (BBa_K206000), and LacI (BBa_C0012).</p><h1 id="1">Week 1 7.12-7.19</h1><p>1. We transformed the BBa_B0015, BBa_C0061, and BBa_B0030 plasmids into E.coli.</p><p>2. We extracted the plasmids and tested them by electrophoresis.</p><p>Results</p><table class="table table-striped"><caption>The concentration of plasmids</caption><thead><tr><td><p>Name</li> | + | <p>Since our team are sorted into 4 groups our target is constructing the Sensor cell in our system. According to our design, we need to construct two gene pathways. Consequently, we use the following parts: pLac (BBa_R0080), Promoter C and arabinose C operon (BBa_I13458), RBS (BBa_B0030), LuxI (BBa_C0061), double terminator (BBa_B0015), Constitutive promoter (BBa_J23100), CinI (BBa_C0076), pBAD (BBa_K206000), and LacI (BBa_C0012).</p><h1 id="1" style="padding-top: 80px; |
| + | margin-top: -45px;">Week 1 7.12-7.19</h1><p>1. We transformed the BBa_B0015, BBa_C0061, and BBa_B0030 plasmids into E.coli.</p><p>2. We extracted the plasmids and tested them by electrophoresis.</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><p>Concentration (ng/3μl)</p> | ||
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</table> | </table> | ||
<p>Note: The different concentrations are from different tubes.</p> | <p>Note: The different concentrations are from different tubes.</p> | ||
| - | <h1 id="2">Week 2 7.20-7.26-</h1> | + | <h1 id="2"style="padding-top: 80px; |
| + | margin-top: -45px;">Week 2 7.20-7.26-</h1> | ||
<p>1. We transformed the BBa_K084012, BBa_I761014, BBa_K081005, BBa_J23100, BBa_K206000, BBa_R0080, BBa_C0076, BBa_C1002, and BBa_I13458 which are tested by cleavage and electrophoresis.</p><p>Results</p><table class="table table-striped"><caption>The concentration of plasmids</caption><thead><tr><td><p>Name</li> | <p>1. We transformed the BBa_K084012, BBa_I761014, BBa_K081005, BBa_J23100, BBa_K206000, BBa_R0080, BBa_C0076, BBa_C1002, and BBa_I13458 which are tested by cleavage and electrophoresis.</p><p>Results</p><table class="table table-striped"><caption>The concentration of plasmids</caption><thead><tr><td><p>Name</li> | ||
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<img src="https://static.igem.org/mediawiki/2014/d/d8/Notebook_of_Sensor%281%291363.png"> | <img src="https://static.igem.org/mediawiki/2014/d/d8/Notebook_of_Sensor%281%291363.png"> | ||
<p>Figure 1. The plasmids that has been cleaved by enzyme.</p> | <p>Figure 1. The plasmids that has been cleaved by enzyme.</p> | ||
| - | <h1 id="3">Week 3 7.27-8.03</h1><p>1. We cleaved the BBa_K084012, BBa_C0012, BBa_I13458 with EcoR I and Spe I, BBa_I761014, and BBa_K206000 with Xba I and Pst I.</p><p>2. We amplified the BBa_K084012 and BBa_K081005 by transforming them into E.coli. And then we extracted the plasmids and tested them by electrophoresis.</p><p>3. We linked the BBa_K084012 and BBa_B0015; BBa_I13458 and BBa_K206000; BBa_K081005 and BBa_C1002 with pSB1A3.</p><p>Results</p><table class="table table-striped"><caption>The concentration of plasmids</caption><thead><tr><td><p>Name</li> | + | <h1 id="3" style="padding-top: 80px; |
| + | margin-top: -45px;">Week 3 7.27-8.03</h1><p>1. We cleaved the BBa_K084012, BBa_C0012, BBa_I13458 with EcoR I and Spe I, BBa_I761014, and BBa_K206000 with Xba I and Pst I.</p><p>2. We amplified the BBa_K084012 and BBa_K081005 by transforming them into E.coli. And then we extracted the plasmids and tested them by electrophoresis.</p><p>3. We linked the BBa_K084012 and BBa_B0015; BBa_I13458 and BBa_K206000; BBa_K081005 and BBa_C1002 with pSB1A3.</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><p>Concentration (ng/3μl)</p> | ||
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<p>The electrophoresis results are shown in Figure 2.</p> | <p>The electrophoresis results are shown in Figure 2.</p> | ||
<img src="https://static.igem.org/mediawiki/2014/2/20/Notebook_of_Sensor%281%292000.png" | <img src="https://static.igem.org/mediawiki/2014/2/20/Notebook_of_Sensor%281%292000.png" | ||
| - | <p>Figure 2. The plasmids that have been cleaved by EcoR I and Spe I or Xba I and Pst I.</p><h1 id="4">Week 4 8.04-8.10</h1><p>1. We re-cleaved the BBa_K081005 for linkage (BBa_K081005 and BBa_C0012 with pSB1A3).</p><p>2. We transformed the linkage products into E.coli. Then we extracted the plasmids to test their qualities.</p><p>3. We transformed, extracted, and tested (BBa_K081005 and BBa_C0012 with pSB1A3).</p><p>4. We sent some of our linkage products to sequence them.</p><p>5. We extracted the BBa_B0015 and BBa_J0465</p><p>6. We cleaved the BBa_B0015 (E,X), BBa_K084012 PLUS BBa_B0015 (E, S), BBa_K084012 (E, S), BBa_K081005 (S,P), and BBa_C1002 (X,P) and we did the gel purification.</p><p>7. We amplified the BBa_I761014 by PCR.</p><p>Results</p><table class="table table-striped"><caption>The concentration of plasmids</caption><thead><tr><td><p>Name</li> | + | <p>Figure 2. The plasmids that have been cleaved by EcoR I and Spe I or Xba I and Pst I.</p><h1 id="4" style="padding-top: 80px; |
| + | margin-top: -45px;">Week 4 8.04-8.10</h1><p>1. We re-cleaved the BBa_K081005 for linkage (BBa_K081005 and BBa_C0012 with pSB1A3).</p><p>2. We transformed the linkage products into E.coli. Then we extracted the plasmids to test their qualities.</p><p>3. We transformed, extracted, and tested (BBa_K081005 and BBa_C0012 with pSB1A3).</p><p>4. We sent some of our linkage products to sequence them.</p><p>5. We extracted the BBa_B0015 and BBa_J0465</p><p>6. We cleaved the BBa_B0015 (E,X), BBa_K084012 PLUS BBa_B0015 (E, S), BBa_K084012 (E, S), BBa_K081005 (S,P), and BBa_C1002 (X,P) and we did the gel purification.</p><p>7. We amplified the BBa_I761014 by PCR.</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</p> | </td><td><p>Concentration</p> | ||
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The electrophoresis results are shown in Figure 3.</p> | The electrophoresis results are shown in Figure 3.</p> | ||
<img src="https://static.igem.org/mediawiki/2014/thumb/a/af/S3.png/580px-S3.png" | <img src="https://static.igem.org/mediawiki/2014/thumb/a/af/S3.png/580px-S3.png" | ||
| - | <p>Figure 3. A and C show the plasmids that have been cleaved. B shows the PCR products.</p><h1 id="5">Week 5 8.11-8.17</h1><p>1. We linked the BBa_K081005 and BBa_C1002; BBa_I13458 PLUS BBa_K206000 and PCR products (Traditional Assembly); BBa_K084012 and BBa_B0015; BBa_K081005 and BBa_C1002 (3A Assembly) and transformed them.</p><p>2. We extracted the plasmids and did the linkage again (3A assembly).</p><p>3. We cleaved the BBa_K084012 (E, S) (Named Cut 1), BBa_B0015 (X, P) (Cut2), BBa_I13458 PLUS BBa_K206000 (E, S) (Cut3), PCR products (X, P) (Cut4), BBa_K81005 (E, S) (Cut5), BBa_C1002 (X, P) (Cut6).</p><p>We linked the Cut1 and Cut2 (Named Lia1); Cut3 and Cut4 (Lia 2 and 3); Cut5 and Cut6 (Lia 4).</li><p><table><tr><td></td> | + | <p>Figure 3. A and C show the plasmids that have been cleaved. B shows the PCR products.</p><h1 id="5" style="padding-top: 80px; |
| + | margin-top: -45px;">Week 5 8.11-8.17</h1><p>1. We linked the BBa_K081005 and BBa_C1002; BBa_I13458 PLUS BBa_K206000 and PCR products (Traditional Assembly); BBa_K084012 and BBa_B0015; BBa_K081005 and BBa_C1002 (3A Assembly) and transformed them.</p><p>2. We extracted the plasmids and did the linkage again (3A assembly).</p><p>3. We cleaved the BBa_K084012 (E, S) (Named Cut 1), BBa_B0015 (X, P) (Cut2), BBa_I13458 PLUS BBa_K206000 (E, S) (Cut3), PCR products (X, P) (Cut4), BBa_K81005 (E, S) (Cut5), BBa_C1002 (X, P) (Cut6).</p><p>We linked the Cut1 and Cut2 (Named Lia1); Cut3 and Cut4 (Lia 2 and 3); Cut5 and Cut6 (Lia 4).</li><p><table><tr><td></td> | ||
</tr><tr><td></td><td></td> | </tr><tr><td></td><td></td> | ||
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The electrophoresis results are shown in Figure 4.</p> | The electrophoresis results are shown in Figure 4.</p> | ||
<img src="https://static.igem.org/mediawiki/2014/9/9b/S4.png" | <img src="https://static.igem.org/mediawiki/2014/9/9b/S4.png" | ||
| - | <p>Figure 4. The parts that have been cleaved for linkage</p><h1 id="6">Week 6 8.18-8.24</h1><p>1. We cleaved the BBa_K081005 (E, S) and BBa_C1002 (X, P) plasmids.</p><p>2. We tested the lia 6-1-1 (X, P), lia 7-1-1 (X, P), lia 7-1-2 (X, P), lia 8-2-1 (E, S), and lia 8-2-2 (E, S) by cleavage and electrophoresis.</p><p>3. We did the gel purification for all the cleavage products.</p><p>4. We cleaved the BBa_K081005 (E, S), lia 6-1-4 (X, P), lia 6-1-1 (X, P), lia 6-1-2 (X, P), lia 6 -1-5 (X, P), lia 4-2-2 (E, S) and control group (E, S). Also, we prepared the linear backbones (pSB1T3 and pSB1C3) by cleavage.</p><p>5. We linked the CUT7 and CUT8 with pSB1T3 (Named NR 1), CUT3 and CUT4 with pSB1C3, pSB1T3, and pSB1K3, BBa_K081005 and CUT6 with pSB1T3 (NR 2).</p><p>6. We transformed the linkage products and extracted the BBa_I761014 and BBa_K084012 plasmids.</p><p>7. We cleaved the BBa_I761014 and BBa_C1002 with Xba I and Pst I. And we tested them by electrophoresis.</p><p>8. We amplified BBa_I761014 by PCR.</p><p>Results</p><p>The concentration of plasmids</p><table><tr><td><p>Name</li> | + | <p>Figure 4. The parts that have been cleaved for linkage</p><h1 id="6" style="padding-top: 80px; |
| + | margin-top: -45px;">Week 6 8.18-8.24</h1><p>1. We cleaved the BBa_K081005 (E, S) and BBa_C1002 (X, P) plasmids.</p><p>2. We tested the lia 6-1-1 (X, P), lia 7-1-1 (X, P), lia 7-1-2 (X, P), lia 8-2-1 (E, S), and lia 8-2-2 (E, S) by cleavage and electrophoresis.</p><p>3. We did the gel purification for all the cleavage products.</p><p>4. We cleaved the BBa_K081005 (E, S), lia 6-1-4 (X, P), lia 6-1-1 (X, P), lia 6-1-2 (X, P), lia 6 -1-5 (X, P), lia 4-2-2 (E, S) and control group (E, S). Also, we prepared the linear backbones (pSB1T3 and pSB1C3) by cleavage.</p><p>5. We linked the CUT7 and CUT8 with pSB1T3 (Named NR 1), CUT3 and CUT4 with pSB1C3, pSB1T3, and pSB1K3, BBa_K081005 and CUT6 with pSB1T3 (NR 2).</p><p>6. We transformed the linkage products and extracted the BBa_I761014 and BBa_K084012 plasmids.</p><p>7. We cleaved the BBa_I761014 and BBa_C1002 with Xba I and Pst I. And we tested them by electrophoresis.</p><p>8. We amplified BBa_I761014 by PCR.</p><p>Results</p><p>The concentration of plasmids</p><table><tr><td><p>Name</li> | ||
</td><td><p>Concentration (ng/3 μl)</p> | </td><td><p>Concentration (ng/3 μl)</p> | ||
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<ing src="https://static.igem.org/mediawiki/2014/thumb/6/61/S5.png/570px-S5.png" | <ing src="https://static.igem.org/mediawiki/2014/thumb/6/61/S5.png/570px-S5.png" | ||
<p>Figure 5. A. gel purification result; B and C. the plasmids that have been cleaved by enzyme. D. PCR results.</p> | <p>Figure 5. A. gel purification result; B and C. the plasmids that have been cleaved by enzyme. D. PCR results.</p> | ||
| - | <h1 id="7">Week 7 8.25-8.31</h1><p>1. We tested the quality of our PCR products by electrophoresis.</p><p>2. We extracted the NR1-1, NR1-2, NR1-3, NR1-4, NR1-5, NR1-6, NR2-1, NR2-2, NR2-3, NR2-4, NR2-5, and NR2-6.</p><p>3. We tested BBa_B0015 (X, P), BBa_I13458 PLUS BBa_K206000 (E, S), BBa_C0012 (S, P), BBa_K084012 (X, P), NR1-1 (X, P), NR1-2 (X, P), NR2-1 (E, S), NR2-2 (E, S) by cleavage.</p><p>4. We cleaved NR2-3, NR2-5, NR2-6 with EcoR I and Spe I.</p><p>5. We linked the BBa_I13458 PLUS BBa_K20600 and our PCR products with pSB1K3 and pSB1C3.</p><p>6. We transformed the linkage products.</p><p>7. We tested the NR1-3, NR1-4 (E, S) and NR2-1/NR2-6 (X, P).</p><p>8. We amplified BBa_I761014 by PCR and did the PCR purification</p><p>9. We extracted NR3 and BBa_I761014 and tested them by electrophoresis and cleavage.</p><p>10. We linked BBa_I13458 PLUS BBa_K206000 and NP2 with pSB1T3 and pSB1C3.</p><p>11. We linked BBa_I13458 PLUS BBa_K206000 and BBa_I761014 with pSB1T3, BBa_0030 and BBa_C0076 with pSB1A3 and pSB1T3.</p><p>Results</p><p>The concentration of plasmids</p><table><tr><td><p>Name</li> | + | <h1 id="7" style="padding-top: 80px; |
| + | margin-top: -45px;">Week 7 8.25-8.31</h1><p>1. We tested the quality of our PCR products by electrophoresis.</p><p>2. We extracted the NR1-1, NR1-2, NR1-3, NR1-4, NR1-5, NR1-6, NR2-1, NR2-2, NR2-3, NR2-4, NR2-5, and NR2-6.</p><p>3. We tested BBa_B0015 (X, P), BBa_I13458 PLUS BBa_K206000 (E, S), BBa_C0012 (S, P), BBa_K084012 (X, P), NR1-1 (X, P), NR1-2 (X, P), NR2-1 (E, S), NR2-2 (E, S) by cleavage.</p><p>4. We cleaved NR2-3, NR2-5, NR2-6 with EcoR I and Spe I.</p><p>5. We linked the BBa_I13458 PLUS BBa_K20600 and our PCR products with pSB1K3 and pSB1C3.</p><p>6. We transformed the linkage products.</p><p>7. We tested the NR1-3, NR1-4 (E, S) and NR2-1/NR2-6 (X, P).</p><p>8. We amplified BBa_I761014 by PCR and did the PCR purification</p><p>9. We extracted NR3 and BBa_I761014 and tested them by electrophoresis and cleavage.</p><p>10. We linked BBa_I13458 PLUS BBa_K206000 and NP2 with pSB1T3 and pSB1C3.</p><p>11. We linked BBa_I13458 PLUS BBa_K206000 and BBa_I761014 with pSB1T3, BBa_0030 and BBa_C0076 with pSB1A3 and pSB1T3.</p><p>Results</p><p>The concentration of plasmids</p><table><tr><td><p>Name</li> | ||
</td><td><p>Concentration (ng/3 μl)</p> | </td><td><p>Concentration (ng/3 μl)</p> | ||
Latest revision as of 03:19, 18 October 2014