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MB Sample ID: SA204514
Local Sample ID: | fur-0-4P |
Subject ID: | SU002216 |
Subject Type: | Bacteria |
Subject Species: | Escherichia coli |
Taxonomy ID: | 562 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002216 |
Subject Type: | Bacteria |
Subject Species: | Escherichia coli |
Taxonomy ID: | 562 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
fur-0-4P | SA204514 | FL025114 | No iron supplementation | Treatment |
Collection:
Collection ID: | CO002209 |
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: | TR002228 |
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 UTI89 mutants. 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: | SP002222 |
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-DLphenylalanine), 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 | AN003485 | AN003486 |
---|---|---|
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: | CH002573 |
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: | MS003246 |
Analysis ID: | AN003485 |
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: | MS003247 |
Analysis ID: | AN003486 |
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 |