"
Team:Bielefeld-CeBiTec/Notebook/Journal/Isobutanol/Jun
From 2014.igem.org
(Difference between revisions)
| Line 48: | Line 48: | ||
<h6><a style="font-size:24px" href="#">Week 1 06/02 - 06/08</a></h6> | <h6><a style="font-size:24px" href="#">Week 1 06/02 - 06/08</a></h6> | ||
</div> | </div> | ||
| - | <div class="content"> | + | <div class="content" style="margin-right:10%; margin-left:10%"> |
</div> | </div> | ||
| Line 65: | Line 65: | ||
<h6><a style="font-size:24px" href="#">Week 2 06/09 - 06/15</a></h6> | <h6><a style="font-size:24px" href="#">Week 2 06/09 - 06/15</a></h6> | ||
</div> | </div> | ||
| - | <div class="content"> | + | <div class="content" style="margin-right:10%; margin-left:10%"> |
</div> | </div> | ||
| Line 82: | Line 82: | ||
<h6><a style="font-size:24px" href="#">Week 3 06/16 - 06/22</a></h6> | <h6><a style="font-size:24px" href="#">Week 3 06/16 - 06/22</a></h6> | ||
</div> | </div> | ||
| - | <div class="content"> | + | <div class="content" style="margin-right:10%; margin-left:10%"> |
</div> | </div> | ||
</div> | </div> | ||
| Line 98: | Line 98: | ||
<h6><a style="font-size:24px" href="#">Week 4 06/23 - 06/29</a></h6> | <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> | <ul> | ||
<li><b><i>alsS</i></b></li> | <li><b><i>alsS</i></b></li> | ||
| Line 169: | Line 169: | ||
<h6><a style="font-size:24px" href="#">Week 5 06/30 - 07/06</a></h6> | <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> | <ul> | ||
<li><b><i>alsS</i></b></li> | <li><b><i>alsS</i></b></li> | ||
Revision as of 10:34, 31 August 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, but not the whole sequence of this part was covered. (VF, VR)
- 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, but not the whole sequence of this part was covered. (VF, VR)
- 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, but not the whole sequence of this part was covered. (VF, VR)
- 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
- K538742: pSB1C3-kivD (parts distribution from 2013, plate 1, 24L)
- Plasmid isolation of kivD
- Sequencing confirmed the identity of pSB1C3-kivD, but not the whole sequence of this part was covered. (VF, VR)
- 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 (1,8 kb)
- 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 (1,55 kb)
- 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 (1,95 kb)
- 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 (1,75 kb)
- 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 (2,2 kb)
- PCR conditions need to be optimized due to low product quality and missing products
