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Team:Bielefeld-CeBiTec/Notebook/Journal/Isobutanol/Jun
From 2014.igem.org
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<td style="padding:30px; width:400px"><h1> June </h1></td> | <td style="padding:30px; width:400px"><h1> June </h1></td> | ||
<td width="100px" align="right"><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Journal/Isobutanol/Jul"><img src="https://static.igem.org/mediawiki/2014/9/9b/Bielefeld-CeBiTec_2014-08-14_Arrow-right.png" width="50px"> </a> </td> | <td width="100px" align="right"><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Journal/Isobutanol/Jul"><img src="https://static.igem.org/mediawiki/2014/9/9b/Bielefeld-CeBiTec_2014-08-14_Arrow-right.png" width="50px"> </a> </td> | ||
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| - | <a style="font-size:24px" href="# | + | <h6><a style="font-size:24px" href="#">Week 4 06/23 - 06/29</a></h6> |
</div> | </div> | ||
| - | <div class="content"> | + | <div class="content" style="margin-right:10%; margin-left:10%"> |
| + | <ul> | ||
| + | <li><b><i>alsS</i></b></li> | ||
| + | <ul> | ||
| + | <li>This week we aim to transform the construct from the parts distribution.</li> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Transformationviaelectroporation" target="_blank">Transformation</a> with electrocompotetent cells</li> | ||
| + | <ul> | ||
| + | <li><a href="http://parts.igem.org/Part:BBa_K539627" target="_blank">K539627</a>: pSB1C3-<i>alsS</i> (parts distribution from 2013, plate 1, 24H)</li> | ||
| + | </ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PurificationPromega" target="_blank">Plasmid isolation</a> of <i>alsS</i></li> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Sequencing" target="_blank">Sequencing</a> confirmed the identity of pSB1C3-<i>alsS</i>.</li> | ||
| + | <li>Glycerin stock created: <i>E. coli</i> KRX pSB1C3-<i>alsS</i></li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | <li><b><i>ilvC</i></b></li> | ||
| + | <ul> | ||
| + | <li>This week we aim to transform the construct from the parts distribution.</li> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Transformationviaelectroporation" target="_blank">Transformation</a> with electrocompotetent cells</li> | ||
| + | <ul> | ||
| + | <li><a href="http://parts.igem.org/Part:BBa_K539621" target="_blank">K539621</a>: pSB1C3-<i>ilvC</i> (parts distribution from 2013, plate 1, 24F)</li> | ||
| + | </ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PurificationPromega" target="_blank">Plasmid isolation</a> of <i>ilC</i></li> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Sequencing" target="_blank">Sequencing</a> confirmed the identity of pSB1C3-<i>ilvC</i>.</li> | ||
| + | <li>Glycerin stock created: <i>E. coli</i> KRX pSB1C3-<i>ilvC</i></li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | <li><b><i>ilvD</i></b></li> | ||
| + | <ul> | ||
| + | <li>This week we aim to transform the construct from the parts distribution.</li> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Transformationviaelectroporation" target="_blank">Transformation</a> with electrocompotetent cells</li> | ||
| + | <ul> | ||
| + | <li><a href="http://parts.igem.org/Part:BBa_K539626" target="_blank">K539626</a>: pSB1C3-<i>ilvD</i> (parts distribution from 2013, plate 1, 22J)</li> | ||
| + | </ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PurificationPromega" target="_blank">Plasmid isolation</a> of <i>ilvD</i></li> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Sequencing" target="_blank">Sequencing</a> confirmed the identity of pSB1C3-<i>ilvD</i>.</li> | ||
| + | <li>Glycerin stock created: <i>E. coli</i> KRX pSB1C3-<i>ilvD</i></li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | <li><b><i>kivD</i></b></li> | ||
| + | <ul> | ||
| + | <li>This week we aim to transform the construct from the parts distribution.</li> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Transformationviaelectroporation" target="_blank">Transformation</a> with electrocompotetent cells</li> | ||
| + | <ul> | ||
| + | <li><a href="http://parts.igem.org/Part:BBa_K539742" target="_blank">K539742</a>: pSB1C3-<i>kivD</i> (parts distribution from 2013, plate 1, 24L)</li> | ||
| + | </ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PurificationPromega" target="_blank">Plasmid isolation</a> of <i>kivD</i></li> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Sequencing" target="_blank">Sequencing</a> confirmed the identity of pSB1C3-<i>kivD</i>.</li> | ||
| + | <li>Glycerin stock created: <i>E. coli</i> KRX pSB1C3-<i>kivD</i></li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | </ul> | ||
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| - | <a style="font-size:24px" href="# | + | <h6><a style="font-size:24px" href="#">Week 5 06/30 - 07/06</a></h6> |
</div> | </div> | ||
| - | <div class="content"> | + | <div class="content" style="margin-right:10%; margin-left:10%"> |
| + | <ul> | ||
| + | <li><b><i>alsS</i></b></li> | ||
| + | <ul> | ||
| + | <li>This week we tried to amplify <i>alsS</i> to use it for the Gibson Assembly.</li> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PCR" target="_blank">PCR amplification</a> (<a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw_alsS_pSB1C3" target="_blank">fw_alsS_pSB1C3</a>, <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv_ilvC_alsS" target="_blank">rv_ilvC_alsS</a>)</li> | ||
| + | <ul> | ||
| + | <li>Annealing temperature: 55 °C</li> | ||
| + | <li>Bands not as expected (~ 1800 bp)</li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | <li><b><i>ilvC</i></b></li> | ||
| + | <ul> | ||
| + | <li>This week we tried to amplify <i>ilvC</i> to use it for the Gibson Assembly.</li> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PCR" target="_blank">PCR amplification</a> (<a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw_alsS_ilvC" target="_blank">fw_alsS_ilvC</a>, <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv_ilvD_ilvC" target="_blank">rv_ilvD_ilvC</a>)</li> | ||
| + | <ul> | ||
| + | <li>Annealing temperature: 55 °C</li> | ||
| + | <li>Bands not as expected (~ 1550 bp)</li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | <li><b><i>ilvD</i></b></li> | ||
| + | <ul> | ||
| + | <li>This week we tried to amplify <i>ilvD</i> to use it for the Gibson Assembly.</li> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PCR" target="_blank">PCR amplification</a> (<a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw_ilvC_ilvD" target="_blank">fw_ilvC_ilvD</a>, <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv_kivD_ilvD" target="_blank">rv_kivD_ilvD</a>)</li> | ||
| + | <ul> | ||
| + | <li>Annealing temperature: 55 °C</li> | ||
| + | <li>Bands not as expected (~ 1950 bp)</li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | <li><b><i>kivD</i></b></li> | ||
| + | <ul> | ||
| + | <li>This week we tried to amplify <i>kivD</i> to use it for the Gibson Assembly.</li> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PCR" target="_blank">PCR amplification</a> (<a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw_ilvD_kivD" target="_blank">fw_ilvD_kivD</a>, <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv_pSB1C3_kivD" target="_blank">rv_pSB1C3_kivD</a>)</li> | ||
| + | <ul> | ||
| + | <li>Annealing temperature: 55 °C</li> | ||
| + | <li>Bands not as expected (~ 1750 bp)</li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | <li><b>Backbone</b></li> | ||
| + | <ul> | ||
| + | <li>This week we tried to amplify the backbone pSB1C3 to use it for the Gibson Assembly.</li> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PCR" target="_blank">PCR amplification</a> (<a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw_kivD_pSB1C3" target="_blank">fw_kivD_pSB1C3</a>, <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv_alsS_pSB1C3" target="_blank">rv_alsS_pSB1C3</a>)</li> | ||
| + | <ul> | ||
| + | <li>Annealing temperature: 55 °C</li> | ||
| + | <li>Bands not as expected (~ 2.200 kb)</li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | <li>PCR conditions need to be optimized due to low product quality and missing products</li> | ||
| + | </ul> | ||
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Latest revision as of 09:12, 17 October 2014
June |
 |
- alsS
- This week we aim to transform the construct from the parts distribution.
- Transformation with electrocompotetent cells
- K539627: pSB1C3-alsS (parts distribution from 2013, plate 1, 24H)
- Plasmid isolation of alsS
- Sequencing confirmed the identity of pSB1C3-alsS.
- Glycerin stock created: E. coli KRX pSB1C3-alsS
- ilvC
- This week we aim to transform the construct from the parts distribution.
- Transformation with electrocompotetent cells
- K539621: pSB1C3-ilvC (parts distribution from 2013, plate 1, 24F)
- Plasmid isolation of ilC
- Sequencing confirmed the identity of pSB1C3-ilvC.
- Glycerin stock created: E. coli KRX pSB1C3-ilvC
- ilvD
- This week we aim to transform the construct from the parts distribution.
- Transformation with electrocompotetent cells
- K539626: pSB1C3-ilvD (parts distribution from 2013, plate 1, 22J)
- Plasmid isolation of ilvD
- Sequencing confirmed the identity of pSB1C3-ilvD.
- Glycerin stock created: E. coli KRX pSB1C3-ilvD
- kivD
- This week we aim to transform the construct from the parts distribution.
- Transformation with electrocompotetent cells
- K539742: pSB1C3-kivD (parts distribution from 2013, plate 1, 24L)
- Plasmid isolation of kivD
- Sequencing confirmed the identity of pSB1C3-kivD.
- Glycerin stock created: E. coli KRX pSB1C3-kivD
- alsS
- This week we tried to amplify alsS to use it for the Gibson Assembly.
- PCR amplification (fw_alsS_pSB1C3, rv_ilvC_alsS)
- Annealing temperature: 55 °C
- Bands not as expected (~ 1800 bp)
- ilvC
- This week we tried to amplify ilvC to use it for the Gibson Assembly.
- PCR amplification (fw_alsS_ilvC, rv_ilvD_ilvC)
- Annealing temperature: 55 °C
- Bands not as expected (~ 1550 bp)
- ilvD
- This week we tried to amplify ilvD to use it for the Gibson Assembly.
- PCR amplification (fw_ilvC_ilvD, rv_kivD_ilvD)
- Annealing temperature: 55 °C
- Bands not as expected (~ 1950 bp)
- kivD
- This week we tried to amplify kivD to use it for the Gibson Assembly.
- PCR amplification (fw_ilvD_kivD, rv_pSB1C3_kivD)
- Annealing temperature: 55 °C
- Bands not as expected (~ 1750 bp)
- Backbone
- This week we tried to amplify the backbone pSB1C3 to use it for the Gibson Assembly.
- PCR amplification (fw_kivD_pSB1C3, rv_alsS_pSB1C3)
- Annealing temperature: 55 °C
- Bands not as expected (~ 2.200 kb)
- PCR conditions need to be optimized due to low product quality and missing products
