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MB Sample ID: SA204481
Local Sample ID: | irp1-0-4P |
Subject ID: | SU002215 |
Subject Type: | Bacteria |
Subject Species: | Escherichia coli |
Taxonomy ID: | 562 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002215 |
Subject Type: | Bacteria |
Subject Species: | Escherichia coli |
Taxonomy ID: | 562 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
irp1-0-4P | SA204481 | FL025113 | standard growth conditions | Treatment |
Collection:
Collection ID: | CO002208 |
Collection Summary: | After 18h of culture, the sample pellet was isolated. The bacterial pellets harvested from 50 mL of broth culture were mixed with 1.2 mL 80% ice-cold methanol (added to internal standard 0.001mg/ml 4-chloro-DL-phenylalanine), then vortexed for 30 s, and placed on dry ice for 30 min. The samples were centrifuged at 18000 × g for 15 min at 4 °C. The frozen samples were ground with beads and the homogenates were centrifuged at 18000 × g for 15 min at 4 °C. The supernatant was mixed with 800μL ice-cold acetonitrile, and then left to stand for 20 minutes in an ice bath. After centrifugation at 18000 × g 4℃ for 15 min, the supernatant was removed and filtered through 0.22μm membrane. |
Sample Type: | Bacterial cells |
Treatment:
Treatment ID: | TR002227 |
Treatment Summary: | M63 medium (1.36% monopotassium phosphate, 0.2% ammonium sulfate, 0.024% magnesium sulfate, 0.001% calcium chloride, and 0.0015% nicotinic acid) was used to form MG1655 and UTI89 mutants. In addition, add ferric chloride solution to the medium to prepare 10μM iron M63 medium, we cultured the wild UTI89 strain in the presence of 10μM iron. The E. coli strain was incubated in LB-agar plate for 12 hours, one colony was isolated to LB broth for further 4 hours incubation, then diluted the solution into M63 medium at a ratio of 1:100 and the cultures were incubated for another18 h at 37°C, 200rpm to culture E. coli. |
Sample Preparation:
Sampleprep ID: | SP002221 |
Sampleprep Summary: | The bacterial pellets harvested from 50 mL of broth culture were mixed with 1.2 mL 80% ice-cold methanol (added to internal standard 0.001mg/ml 4-chloro-DL-phenylalanine), then vortexed for 30 s, and placed on dry ice for 30 min. The samples were centrifuged at 18000 × g for 15 min at 4 °C. The frozen samples were ground with beads and the homogenates were centrifuged at 18000 × g for 15 min at 4 °C. The supernatant was mixed with 800μL ice-cold acetonitrile, and then left to stand for 20 minutes in an ice bath. After centrifugation at 18000 × g 4℃ for 15 min, the supernatant was removed and filtered through 0.22μm membrane. For LC/MS based metabolomics analysis, the dried samples were dissolved in 200μL water and 5μL aliquots were analyzed. |
Combined analysis:
Analysis ID | AN003483 | AN003484 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Agilent 1290 Infinity | Agilent 1290 Infinity |
Column | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
MS Type | ESI | ESI |
MS instrument type | Triple quadrupole | Triple quadrupole |
MS instrument name | Agilent 6495 QQQ | Agilent 6495 QQQ |
Ion Mode | POSITIVE | NEGATIVE |
Units | peak area | peak area |
Chromatography:
Chromatography ID: | CH002572 |
Instrument Name: | Agilent 1290 Infinity |
Column Name: | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003244 |
Analysis ID: | AN003483 |
Instrument Name: | Agilent 6495 QQQ |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | Agilent MassHunter Workstation Data Acquisition Agilent MassHunter QualitativeAnalysis B.07.00 Agilent MassHunter Quantitative Analysis (for QQQ) |
Ion Mode: | POSITIVE |
MS ID: | MS003245 |
Analysis ID: | AN003484 |
Instrument Name: | Agilent 6495 QQQ |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | Agilent MassHunter Workstation Data Acquisition Agilent MassHunter QualitativeAnalysis B.07.00 Agilent MassHunter Quantitative Analysis (for QQQ) |
Ion Mode: | NEGATIVE |