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Team:Bielefeld-CeBiTec/Notebook/Journal/rMFC/Sep
From 2014.igem.org
(Difference between revisions)
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<ul> | <ul> | ||
<li><b><i>FumA</i></b></li> | <li><b><i>FumA</i></b></li> | ||
| + | <ul> | ||
| + | <li> This week we assembled <i>FumA</i> with different constitutive promotors</li> | ||
<ul> | <ul> | ||
<li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#RestrictionDigestion" target="_blank">Restriction digestion</a> with <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/KitsAndEnzymes#RestrictionEnzymes" target="_blank"><i>EcoR</i>I</a> and <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/KitsAndEnzymes#RestrictionEnzymes" target="_blank"><i>Pst</i>I</a> </li> | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#RestrictionDigestion" target="_blank">Restriction digestion</a> with <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/KitsAndEnzymes#RestrictionEnzymes" target="_blank"><i>EcoR</i>I</a> and <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/KitsAndEnzymes#RestrictionEnzymes" target="_blank"><i>Pst</i>I</a> </li> | ||
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<li>Bands as expected (~2070 bp and ~1800 bp)</li> | <li>Bands as expected (~2070 bp and ~1800 bp)</li> | ||
</ul> | </ul> | ||
| + | </ul> | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
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<ul> | <ul> | ||
<li><b><i>GSU 3274</i></b></li> | <li><b><i>GSU 3274</i></b></li> | ||
| + | <ul><li> This week we assembled <i>GSU 3274</i> with different promotors</li> | ||
<ul> | <ul> | ||
<li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#BioBrick" target="_blank">BioBrick Assembly</a> (Suffix)</li> | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#BioBrick" target="_blank">BioBrick Assembly</a> (Suffix)</li> | ||
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</ul> | </ul> | ||
</ul> | </ul> | ||
| - | + | </ul> | |
| - | + | </ul> | |
| + | <br> | ||
| + | <ul> | ||
| + | <li><b>Electrobiochemical reactor system</b></li> | ||
| + | <ul> | ||
| + | <li>This week we arranged the complete <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/rMFC/Construction">H-cell reactor </a> for the first time to start first trials. Therefore we prepare the<a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Media#H-cell%20buffer" target="_blank"> H-cell buffers</a> and the <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Nafion%C2%AE%20membrane" target="_blank">Nafion® membrane</a> which ensures the division of the two compartments.</li> | ||
| + | |||
| + | </ul> | ||
| + | </ul> | ||
| + | <br> | ||
| + | <ul> | ||
| + | <li><b>NAD/NADH Assay</b></li> | ||
| + | <ul> | ||
| + | <li>This week we startet the first test with the <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PromegaGlo%E2%84%A2%20Assay" target="_blank">Promega NAD/NADH-Glo™ Assay</a> to evaluate later cultivations respectively.</li> | ||
| + | |||
| + | </ul> | ||
| + | </ul> | ||
| + | <br> | ||
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</div> | </div> | ||
<div class="content" style="margin-right:10%; margin-left:10%"> | <div class="content" style="margin-right:10%; margin-left:10%"> | ||
| + | <ul> | ||
| + | <li><b><i>ccm</i></b></li> | ||
| + | <ul> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PurificationPromega" target="_blank">Plasmid isolation</a> of pSB1C3_<i>ccm</i></li> | ||
| + | </ul> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#RestrictionDigestion" target="_blank">Restriction digestion</a> with <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/KitsAndEnzymes#RestrictionEnzymes" target="_blank"><i>Not</i>I</a></li> | ||
| + | <ul> | ||
| + | <li>Bands not as expected (~6230 bp)</li> | ||
| + | </ul> | ||
| + | </ul> | ||
</div> | </div> | ||
</div> | </div> | ||
Revision as of 11:38, 14 October 2014
 |
September |
 |
- FumA
- This week we assembled FumA with different constitutive promotors
- Restriction digestion with EcoRI and PstI
- Bands as expected (~2070 bp and ~1800 bp) for pSB1C3_T7_FumA
- BioBrick Assembly (Suffix)
- Transformation of all constructs with electrocompotetent cells
- Colony PCR (VF-Primer, VR-Primer)
- Annealing temperature: 55 °C
- Bands as expected (~2100 bp) for all constructs
- Plasmid isolation of the positive colonies
- Restriction digestion with EcoRI and PstI
- Bands as expected (~2070 bp and ~1800 bp)
- GSU 3274
- This week we assembled GSU 3274 with different promotors
- BioBrick Assembly (Suffix)
- Transformation of all constructs with electrocompotetent cells
- Colony PCR (VF-Primer, VR-Primer)
- Annealing temperature: 55°C
- Bands as expected (~ bp) for pSB1C3_ptac and pSB1A2_T7
- Electrobiochemical reactor system
- This week we arranged the complete H-cell reactor for the first time to start first trials. Therefore we prepare the H-cell buffers and the Nafion® membrane which ensures the division of the two compartments.
- NAD/NADH Assay
- This week we startet the first test with the Promega NAD/NADH-Glo™ Assay to evaluate later cultivations respectively.
- ccm
- Plasmid isolation of pSB1C3_ccm
- Restriction digestion with NotI
- Bands not as expected (~6230 bp)
