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Team:Bielefeld-CeBiTec/Notebook/Journal/Isobutanol/Jun
From 2014.igem.org
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June |
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- 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
