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Team:Bielefeld-CeBiTec/Notebook/Journal/Isobutanol/Jun
From 2014.igem.org
(Difference between revisions)
| Line 184: | Line 184: | ||
<li><b><i>alsS</i></b></li> | <li><b><i>alsS</i></b></li> | ||
<ul> | <ul> | ||
| - | <li>This week we tried to aplify <i>alsS</i>.</li> | + | <li>This week we tried to aplify <i>alsS</i> to use it for the Gibson Assembly.</li> |
<ul> | <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> | <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> | ||
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<li><b><i>ilvC</i></b></li> | <li><b><i>ilvC</i></b></li> | ||
<ul> | <ul> | ||
| - | <li>This week we tried to aplify <i>ilvC</i>.</li> | + | <li>This week we tried to aplify <i>ilvC</i> to use it for the Gibson Assembly.</li> |
<ul> | <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> | <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> | ||
| Line 206: | Line 206: | ||
<li><b><i>ilvD</i></b></li> | <li><b><i>ilvD</i></b></li> | ||
<ul> | <ul> | ||
| - | <li>This week we tried to aplify <i>ilvD</i>.</li> | + | <li>This week we tried to aplify <i>ilvD</i> to use it for the Gibson Assembly.</li> |
<ul> | <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> | <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> | ||
Revision as of 15:06, 30 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 the identity of pSB1C3-kivD, but not the whole sequence of this part 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 the identity of pSB1C3-alsS, but not the whole sequence of this part 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 the identity of pSB1C3-ilvC, but not the whole sequence of this part 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 the identity of pSB1C3-ilvD, but not the whole sequence of this part was covered. (VF and VR)
- Glycerin stock created: E. coli KRX pSB1C3-ilvD
- kivD
- This week we tried to aplify kivD.
- PCR amplification (fw_ilvD_kivD, rv_pSB1C3_kivD)
- Elongation time: 60s
- Bands not as expected (1,75 kb)
- alsS
- This week we tried to aplify alsS to use it for the Gibson Assembly.
- PCR amplification (fw_alsS_pSB1C3, rv_ilvC_alsS)
- Elongation time: 60s
- Bands not as expected (1,8 kb)
- ilvC
- This week we tried to aplify ilvC to use it for the Gibson Assembly.
- PCR amplification (fw_alsS_ilvC, rv_ilvD_ilvC)
- Elongation time: 60s
- Bands not as expected (1,55 kb)
- ilvD
- This week we tried to aplify ilvD to use it for the Gibson Assembly.
- PCR amplification (fw_ilvC_ilvD, rv_kivD_ilvD)
- Elongation time: 60s
- Bands not as expected (1,95 kb)
- Backbone
- This week we tried to aplify the backbone pSB1C3 to use it for the Gibson Assembly.
- PCR amplification (fw_kivD_pSB1C3, rv_alsS_pSB1C3)
- Elongation time: 60s
- Bands not as expected (2,2 kb)
- PCR conditions need to be optimized due to low product quality and missing products
