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Team:Bielefeld-CeBiTec/Notebook/Journal/CO2-fixation/Sep
From 2014.igem.org
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</ul><!-- /T7 Hneap --> | </ul><!-- /T7 Hneap --> | ||
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| + | <ul><!-- sap --> | ||
| + | <li><b><i>sap</i></b></li> | ||
| + | <ul> | ||
| + | <li>We tried to assemble both parts of <i>sap</i>.</li> | ||
| + | </ul> | ||
| + | </ul> | ||
</div> | </div> | ||
</div> | </div> | ||
Revision as of 15:09, 30 September 2014
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September |
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- csoS1-4
- We tried to amplify csoS1-4 (shell proteins) for a fusion with GFP.
- PCR amplification (fw-GFP-csoS1B, rv-rv-GFP-csoS1A)
- Annealing temperature: 54 °C
- Bands as expected (~4000 bp (~1800 bp csoS1-4, ~2200 bp pSB1C3))
- PCR products were purified
- csoS1-4 and GFP
- We tried to assemble the shell proteins of the carboxysome and GFP.
- Gibson Assembly with pSB1C3_csoS1-4 and GFP
- Transformation with electrocompotetent cells
- Colony PCR (fw-csoS1A-GFP, rv-csoS1B-GF)
- Annealing temperature: 68 °C
- Bands as expected (~1000 bp)
- Plasmid isolation of pSB1C3_csoS1-4_GFP
- Restriction digestion with NotI
- Bands (as expected (~2000 bp and ~2400 bp)
- csoS1-4_GFP and T7
- We tried to assemble our pSB1C3_csoS1-4_GFP construct with the T7 promotor.
- BioBrick Assembly (Suffix)
- Transformation with electrocompotetent cells
- Colony PCR (VF-Primer, rv_csoS4A_PstI)
- Annealing temperature: 55 °C
- Bands as expected (~320 bp)
- can and csoS1-4
- This week we tried to find positive clones of our transformation.
- Colony PCR (VF-Primer, VR-Primer)
- Annealing temperature: 55 °C
- Bands as expected (~3600 bp)
- Plasmid isolation of pSB1C3_can_csoS1-4
- Restriction digestion with EcoRI and PstI
- Bands as expected (~2100 bp and ~3300 bp)
- can_csoS1-4 and csoS1D
- We tried to assemble our pSB1C3_can_csoS1-4 construct with csoS1D.
- BioBrick Assembly (Suffix)
- Transformation with electrocompotetent cells
- Colony PCR (VF-Primer, VR-Primer)
- Annealing temperature: 55 °C
- Bands as expected (~4300 bp)
- Plasmid isolation of pSB1C3_can_csoS1-4_csoS1D
- Restriction digestion with NotI
- Bands as expected (~2000 bp and ~4000 bp)
- Hneap and T7
- This week we tried to assemble the T7 promotor with the Hneap.
- BioBrick Assembly (Suffix)
- Transformation with electrocompotetent cells
- Hneap and T7
- We tried to find positiv clones of pSB3A2_T7_Hneap.
- Colony PCR (VF-Primer, VR-Primer)
- Annealing temperature: 55 °C
- Bands as expected (~2000 bp)
- Plasmid isolation of pSB1A2_T7_Hneap
- Restriction digestion with EcoRI and PstI
- Bands as expected (~2000 bp and ~1800 bp)
- sap
- We tried to assemble both parts of sap.
