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Team:Bielefeld-CeBiTec/Notebook/Journal/CO2-fixation/Oct
From 2014.igem.org
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October |
- tkt
- This week we wanted to do the enzyme assay for the SBPase.
- The purified enzyme from His-Tag purification werde concentrated and desalted with Amicon Ultra-15 with 10kD cut-off
- fba
- This week we wanted to do the enzyme assay for the SBPase.
- The purified enzyme from His-Tag purification werde concentrated and desalted with Amicon Ultra-15 with 10kD cut-off
- glpX
- This week we wanted to do the enzyme assay for the SBPase.
- The purified enzyme from His-Tag purification werde concentrated and desalted with Amicon Ultra-15 with 10kD cut-off
- prkA and ptac_Hneap
- We tried to bring the prkA with RBS behind our construct ptac_Hneap but it was not successful.
- prkA
- We tried to bring our prkA with RBS in pSB1C3.
- BioBrick Assembly (Suffix)
- Transformation with chemocompetent cells
- Plasmid isolation of [Construct]
- Restriction digestion with PstI and EcoRI
- Bands as expected (~2000 bp and ~1100 bp)
- ptacglpX
- We made a cultivation with our construct pSB1C3_ptac_glpX in xylose and glucose to see if there is a difference between the two media and also between induced and not induced. As a reference we used the E. coli wildtype. Because the culture in xylose did not grow we only could measure the OD600 of the glucose cultures. We used 30 ml in 250 ml flasks a 37°C and 250 rpm. The induction was with IPTG. It lasted 12 hours. We made 2 biological and 2 technical replica for each culture. We took two samples (1 ml) of each biological replica for HPLC analysis that followed.
- ptacglpX
- We made a cultivation with our construct pSB1C3_ptac_glpX in xylose to see if there is a difference between this cultivation and the one with glucose and also between induced and not induced. As a reference we used the E. coli wildtype. We used 30 ml in 250 ml flasks a 37°C and 250 rpm. The induction was with IPTG. It lasted 15 hours. We made 2 biological and 2 technical replica for each culture. We took two samples (1 ml) of each biological replica for HPLC analysis that followed.
