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