(reporterkram rein) |
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<div class="labWeek"><h5>Lab Week 17: <div class="week_date"> August 8 - August 14</div></h5></div></td> | <div class="labWeek"><h5>Lab Week 17: <div class="week_date"> August 8 - August 14</div></h5></div></td> | ||
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | <td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | ||
− | <td><p> | + | <td><p><h6>Reporter System</h6> |
+ | <b>BBa_K1976002:</b><br> | ||
+ | The mVenus-LVA-gene was synthesized by idt and amplified through PCR using the SP-amplify-fw-Rep1 (fw) and SP-amplify-rev-Rep2 (REV) oligonucleotides.</p> | ||
+ | |||
</td></tr> | </td></tr> | ||
<tr><td></td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/7/7c/T--TU_Darmstadt--iconTech.png"/></td> | <tr><td></td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/7/7c/T--TU_Darmstadt--iconTech.png"/></td> | ||
Line 278: | Line 281: | ||
<div class="labWeek"><h5>Lab Week 18: <div class="week_date"> August 15 - August 21</div></h5></div></td> | <div class="labWeek"><h5>Lab Week 18: <div class="week_date"> August 15 - August 21</div></h5></div></td> | ||
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | <td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | ||
− | <td><p> | + | <td><p><h6>Reporter System</h6> |
+ | <b>BBa_K1976002:</b><br> | ||
+ | The PCR product was verified by agarose gel electrophoresis and subsequently purified. The mVenus amplicon was cut with EcoRI and PstI and ligated into a pSB1C3 vector using T4-ligase. </p> | ||
+ | |||
</td></tr> | </td></tr> | ||
<tr><td></td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/7/7c/T--TU_Darmstadt--iconTech.png"/></td> | <tr><td></td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/7/7c/T--TU_Darmstadt--iconTech.png"/></td> | ||
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<div class="labWeek"><h5>Lab Week 19: <div class="week_date"> August 22 - August 28</div></h5></div></td> | <div class="labWeek"><h5>Lab Week 19: <div class="week_date"> August 22 - August 28</div></h5></div></td> | ||
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | <td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | ||
− | <td><p> | + | <td><p><h6>Reporter System</h6> |
+ | <b>BBa_K1976002:</b><br> | ||
+ | The ligation product was transformed into E.coli Top 10 by heat shock transformation and the cells were spread out on a CMP agar plate.</p> | ||
+ | |||
</td></tr> | </td></tr> | ||
<tr><td></td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/7/7c/T--TU_Darmstadt--iconTech.png"/></td> | <tr><td></td><td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/7/7c/T--TU_Darmstadt--iconTech.png"/></td> | ||
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<div class="labWeek"><h5>Lab Week 20: <div class="week_date"> August 29 - September 4</div></h5></div></td> | <div class="labWeek"><h5>Lab Week 20: <div class="week_date"> August 29 - September 4</div></h5></div></td> | ||
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | <td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | ||
+ | <td><p><h6>Reporter System</h6> | ||
+ | <b>BBa_K1976002:</b><br> | ||
+ | Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones. </p> | ||
+ | |||
+ | </td></tr> | ||
+ | |||
<td><p><h6>Metabolic burden</h6> | <td><p><h6>Metabolic burden</h6> | ||
<b>BBa_K1976001:</b><br> | <b>BBa_K1976001:</b><br> | ||
Line 325: | Line 340: | ||
<div class="labWeek"><h5>Lab Week 21: <div class="week_date"> September 5 - September 11</div></h5></div></td> | <div class="labWeek"><h5>Lab Week 21: <div class="week_date"> September 5 - September 11</div></h5></div></td> | ||
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | <td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | ||
+ | <td><p><h6>Reporter System</h6> | ||
+ | <b>BBa_K1976002:</b><br> | ||
+ | Isolated Plasmid DNA Sequence was verified by Sanger Sequencing. </p> | ||
+ | </td></tr> | ||
<td><p><h6>Metabolic burden</h6> | <td><p><h6>Metabolic burden</h6> | ||
<b>BBa_K1976001:</b><br> | <b>BBa_K1976001:</b><br> | ||
Line 342: | Line 361: | ||
<div class="labWeek"><h5>Lab Week 22: <div class="week_date"> September 12 - September 18</div></h5></div></td> | <div class="labWeek"><h5>Lab Week 22: <div class="week_date"> September 12 - September 18</div></h5></div></td> | ||
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | <td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | ||
+ | <td><p><h6>Reporter System</h6> | ||
+ | <b>BBa_K1976003:</b><br> | ||
+ | The mVenus-LVA-pSB1C3 plasmid was cut with <i>EcoRI</i> and <i>PstI</i> and ligated into a T7-RBS-pSB1A3 plasmid which was cut with <i>SpeI</i> and <i>PstI.</i> </p> | ||
+ | </td></tr> | ||
+ | |||
<td><p><h6>Metabolic burden</h6> | <td><p><h6>Metabolic burden</h6> | ||
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<div class="labWeek"><h5>Lab Week 23: <div class="week_date"> September 19 - September 25</div></h5></div></td> | <div class="labWeek"><h5>Lab Week 23: <div class="week_date"> September 19 - September 25</div></h5></div></td> | ||
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | <td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | ||
+ | <td><p><h6>Reporter System</h6> | ||
+ | <b>BBa_K1976003:</b><br> | ||
+ | The ligation product was transformed into <i>E.coli</i> Top 10 by heat shock transformation and the cells were spread out on a AMP agar plate. </p> | ||
+ | </td></tr> | ||
+ | |||
+ | |||
<td><p><h6>Metabolic burden</h6> | <td><p><h6>Metabolic burden</h6> | ||
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<div class="labWeek"><h5>Lab Week 24: <div class="week_date"> September 26 - October 2</div></h5></div></td> | <div class="labWeek"><h5>Lab Week 24: <div class="week_date"> September 26 - October 2</div></h5></div></td> | ||
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | <td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | ||
+ | <td><p><h6>Reporter System</h6> | ||
+ | <b>BBa_K1976003:</b><br> | ||
+ | Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones . </p> | ||
+ | |||
+ | </td></tr> | ||
<td><p><h6>Metabolic burden</h6> | <td><p><h6>Metabolic burden</h6> | ||
Line 411: | Line 446: | ||
<div class="labWeek"><h5>Lab Week 25: <div class="week_date"> October 3 - October 9</div></h5></div></td> | <div class="labWeek"><h5>Lab Week 25: <div class="week_date"> October 3 - October 9</div></h5></div></td> | ||
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | <td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | ||
+ | <td><p><h6>Reporter System</h6> | ||
+ | <b>BBa_K1976003:</b><br> | ||
+ | The isolated Plasmid DNA was verified by Sanger sequencing.</p> | ||
+ | </td></tr> | ||
<td><p><h6>Metabolic burden</h6> | <td><p><h6>Metabolic burden</h6> | ||
Line 429: | Line 468: | ||
<div class="labWeek"><h5>Lab Week 26: <div class="week_date"> October 10 - October 16</div></h5></div></td> | <div class="labWeek"><h5>Lab Week 26: <div class="week_date"> October 10 - October 16</div></h5></div></td> | ||
<td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | <td><img class="labjournalIcon" src="https://static.igem.org/mediawiki/2016/b/b5/T--TU_Darmstadt--iconLab3.png"/></td> | ||
+ | <td><p><h6>Reporter System</h6> | ||
+ | <b>BBa_K1976003:</b><br> | ||
+ | Subsequently the psB1A3 vector containing brick was cut with <i>EcoRI</i> and <i>PstI</i> and ligated into a pSB1C3 vector using T4‑Ligase.</p> | ||
+ | </td></tr> | ||
<td><p><h6>Metabolic burden</h6> | <td><p><h6>Metabolic burden</h6> | ||
Revision as of 17:02, 19 October 2016
![iGEM TU Darmstadt 2016](https://static.igem.org/mediawiki/2016/8/83/T--TU_Darmstadt--titel.png)
NOTEBOOK
Labjournal 2016
For half a year we were working in the wet lab , developed a pipetting robot
and simulated the molecular dynamics of our proteins
.
As well we would like to highlight certain social events
which helped us to further improve our team spirit.
Lab Week 1: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 2: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 3: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 4: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 5: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 6: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 7: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 8: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 9: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 10: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 11: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 12: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 13: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 14: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 15: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 16: |
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Hier könnte Ihr labjournal stehen! |
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robotics | |
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modeling | |
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social |
Lab Week 17: |
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Reporter SystemBBa_K1976002:The mVenus-LVA-gene was synthesized by idt and amplified through PCR using the SP-amplify-fw-Rep1 (fw) and SP-amplify-rev-Rep2 (REV) oligonucleotides. |
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robotics | |
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modeling | |
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social |
Lab Week 18: |
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Reporter SystemBBa_K1976002:The PCR product was verified by agarose gel electrophoresis and subsequently purified. The mVenus amplicon was cut with EcoRI and PstI and ligated into a pSB1C3 vector using T4-ligase. |
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robotics | |
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modeling | |
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social |
Lab Week 19: |
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Reporter SystemBBa_K1976002:The ligation product was transformed into E.coli Top 10 by heat shock transformation and the cells were spread out on a CMP agar plate. |
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robotics | |
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modeling | |
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social |
Lab Week 20: |
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Reporter SystemBBa_K1976002:Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones. | Metabolic burdenBBa_K1976001:The BioBrick BBa_I11020 was synthezised with a LVA‑tag and with B0034 as a ribosome binding site by IDT. |
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robotics | |
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modeling | |
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social |
Lab Week 21: |
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Reporter SystemBBa_K1976002:Isolated Plasmid DNA Sequence was verified by Sanger Sequencing. | Metabolic burdenBBa_K1976001:The LVA‑tag was deleted by PCR with the primer Del_LVA_fw and Del_LVA_rev. |
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robotics | |
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modeling | |
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social |
Lab Week 22: |
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Reporter SystemBBa_K1976003:The mVenus-LVA-pSB1C3 plasmid was cut with EcoRI and PstI and ligated into a T7-RBS-pSB1A3 plasmid which was cut with SpeI and PstI. | Metabolic burdenBBa_K1976000:The BioBrick BBa_E0240 was cut with the restriction enzymes XbaI and PstI. BBa_J61002 was cut with SpeI and PstI. The two restriction products were ligated by standard T4‑Ligation protocol. BBa_K1976001: B0034‑BBa_I11020 was cut with XbaI and PstI. BBa_J23101 was cut with SpeI and PstI. Both fragments were ligated using the T4‑Ligase by standard protocol. |
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robotics | |
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modeling | |
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social |
Lab Week 23: |
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Reporter SystemBBa_K1976003:The ligation product was transformed into E.coli Top 10 by heat shock transformation and the cells were spread out on a AMP agar plate. | Metabolic burdenBBa_K1976000:After plasmid preparation, the plasmid was cut with XbaI and PstI. We had the attp‑site BBa_11023 flanked by the two additional bidirectional transcription terminators BBa_1001 synthesized by IDT. The attp‑site of BBa_11023 was not the original λ‑attp‑site. Instead the sequence corresponded to the attP2‑site used in the Gateway ® cloning system. The construct was cut with EcoRI and SpeI. The pSB1C3 backbone was cut with EcoRI and PstI. Together with the previously cut BBa_J61002‑BBa_E0240 construct the synthesized BBa_1001‑BBa_11023‑BBa_1001 construct was ligated in the cut pSB1C3. BBa_K1976001: The ligation product was transformed into E. coli Top 10 via heatshock. |
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robotics | |
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modeling | |
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social |
Lab Week 24: |
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Reporter SystemBBa_K1976003:Clones were screened with colony PCR using VF2 and VR oligonucleotides. Plasmid DNA was isolated from positive clones . | Metabolic burdenBBa_K1976000:The attP2‑site was mutated to λ‑attp via Quick‑Change‑PCR, using the following primers: QC_attp_fw and QC_attp_rev (for more details see our primerlist). BBa_K1976001: To check whether BBa_I11020 was expressed properly a SDS‑Page was made. |
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robotics | |
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modeling | |
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social |
Lab Week 25: |
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Reporter SystemBBa_K1976003:The isolated Plasmid DNA was verified by Sanger sequencing. | Metabolic burdenBBa_K1976000:The LVA‑tag was added by PCR with the overhang primer LVA_GFP_fw and LVA_GFP_rev. |
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modeling | |
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Lab Week 26: |
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Reporter SystemBBa_K1976003:Subsequently the psB1A3 vector containing brick was cut with EcoRI and PstI and ligated into a pSB1C3 vector using T4‑Ligase. | Metabolic burdenBBa_K1976000:For plasmid curing the final plasmid construct and the low copy backbone pSB4A5 were cut with EcoRI and PstI and then ligated. BBa_K1976001: For plasmid curing the final plasmid construct and the low copy vector pSB4K5 were cut with EcoRI and PstI and then ligated. BBa_K1976000 and BBa_K1976001 were cotransformed into E. coli Top 10. To check whether the genomic integration was successful a cPCR with attb_fw and VR primers was made. |
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Lab Week 27: |
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Metabolic burdenPlasmid curing could not be performed due to lack of time. |
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