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Team:Bielefeld-CeBiTec/Notebook/Journal/Isobutanol/Jun
From 2014.igem.org
(Difference between revisions)
| Line 104: | Line 104: | ||
<li>This week we aim to transform the construct from the parts distribution.</li> | <li>This week we aim to transform the construct from the parts distribution.</li> | ||
<ul> | <ul> | ||
| - | <li>Transformation with electrocompotetent cells</li> | + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Transformationviaelectroporation" target="_blank">Transformation</a> with electrocompotetent cells</li> |
<ul> | <ul> | ||
<li>K538742: <i>pSB1C3-kivD</i> (parts distribution from 2013, plate 1, 24L)</li> | <li>K538742: <i>pSB1C3-kivD</i> (parts distribution from 2013, plate 1, 24L)</li> | ||
| Line 117: | Line 117: | ||
<li>This week we aim to transform the construct from the parts distribution.</li> | <li>This week we aim to transform the construct from the parts distribution.</li> | ||
<ul> | <ul> | ||
| - | <li>Transformation with electrocompotetent cells</li> | + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Transformationviaelectroporation" target="_blank">Transformation</a> with electrocompotetent cells</li> |
<ul> | <ul> | ||
<li>K539627: pSB1C3-<i>alsS</i> (parts distribution from 2013, plate 1, 24H)</li> | <li>K539627: pSB1C3-<i>alsS</i> (parts distribution from 2013, plate 1, 24H)</li> | ||
| Line 130: | Line 130: | ||
<li>This week we aim to transform the construct from the parts distribution.</li> | <li>This week we aim to transform the construct from the parts distribution.</li> | ||
<ul> | <ul> | ||
| - | <li>Transformation with electrocompotetent cells</li> | + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Transformationviaelectroporation" target="_blank">Transformation</a> with electrocompotetent cells</li> |
<ul> | <ul> | ||
<li>K539621: pSB1C3-<i>ilvC</i> (parts distribution from 2013, plate 1, 24F)</li> | <li>K539621: pSB1C3-<i>ilvC</i> (parts distribution from 2013, plate 1, 24F)</li> | ||
| Line 143: | Line 143: | ||
<li>This week we aim to transform the construct from the parts distribution.</li> | <li>This week we aim to transform the construct from the parts distribution.</li> | ||
<ul> | <ul> | ||
| - | <li>Transformation with electrocompotetent cells</li> | + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Transformationviaelectroporation" target="_blank">Transformation</a> with electrocompotetent cells</li> |
<ul> | <ul> | ||
<li>K539626: pSB1C3-<i>ilvD</i> (parts distribution from 2013, plate 1, 22J)</li> | <li>K539626: pSB1C3-<i>ilvD</i> (parts distribution from 2013, plate 1, 22J)</li> | ||
Revision as of 12:57, 27 August 2014
 |
June |
 |
- 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 identities of all parts, but not whole sequence of the parts was covered. (VF and VR)
- Glycerin stock created: E. coli KRX pSB1C3-kivD
- 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 identities of all parts, but not whole sequence of the parts was covered. (VF and 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 identities of all parts, but not whole sequence of the parts was covered. (VF and 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 identities of all parts, but not whole sequence of the parts was covered. (VF and VR)
- Glycerin stock created: E. coli KRX pSB1C3-ilvD
- kivD
- This week we tried to combine kivD with the other using the Gibson Assembly.
- PCR amplification (fw_ilvD_kivD, rv_pSB1C3_kivD)
- Elongation time: 90s
- Bands (not) as expected (... bp)
- Gibson Assembly with kivD, alsS, ilvC and ilvD on pSB1C3
- alsS
- This week we tried to combine alsS with the other using the Gibson Assembly.
- PCR amplification (fw_alsS_pSB1C3, rv_ilvC_alsS)
- Elongation time: 90s
- Bands (not) as expected (... bp)
- Gibson Assembly with kivD, alsS, ilvC and ilvD on pSB1C3
- ilvC
- This week we tried to combine ilvC with the other using the Gibson Assembly.
- PCR amplification (fw_alsS_ilvC, rv_ilvD_ilvC)
- Elongation time: 90s
- Bands (not) as expected (... bp)
- Gibson Assembly with kivD, alsS, ilvC and ilvD on pSB1C3
- ilvD
- This week we tried to combine ilvD with the other using the Gibson Assembly.
- PCR amplification (fw_ilvC_ilvD, rv_kivD_ilvD)
- Elongation time: 90s
- Bands (not) as expected (... bp)
- Gibson Assembly with kivD, alsS, ilvC and ilvD on pSB1C3
- Backbone
- This week we amplified the backbone to use it for the Gibson Assembly.
- PCR amplification (fw_kivD_pSB1C3, rv_alsS_pSB1C3)
- Elongation time: 90s
- Bands (not) as expected (... bp)
- Gibson Assembly with kivD, alsS, ilvC and ilvD on pSB1C3
- PCR conditions need to be optimized due to low product quality and missing products
