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MB Sample ID: SA204444
Local Sample ID: | MG1655-10-1S |
Subject ID: | SU002214 |
Subject Type: | Bacteria |
Subject Species: | Escherichia coli |
Taxonomy ID: | 562 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002214 |
Subject Type: | Bacteria |
Subject Species: | Escherichia coli |
Taxonomy ID: | 562 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
MG1655-10-1S | SA204444 | FL025110 | 10 μM iron supplementation | Treatment |
Collection:
Collection ID: | CO002207 |
Collection Summary: | After 18h of culture, the sample supernatant was isolated.Then, 2μL 0.1M ferric chloride was mixed with 2 mL of cell supernatant. After incubating at room temperature for 15 minutes, the precipitate was removed by centrifugation at 20000 × g for 15 min at 4 °C. The supernatant was added to an SPE plate (Waters, Oasis HLB) and washed with 0.5 mL 5% methanol, and then eluted with 0.5 mL 100% methanol to obtain the siderophores. |
Sample Type: | Bacterial cells |
Treatment:
Treatment ID: | TR002226 |
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. In addition, add ferric chloride solution to the medium to prepare 10μM iron M63 medium, we cultured the wild UTI89 strain and MG1655 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: | SP002220 |
Sampleprep Summary: | Siderophores were extracted as previously described. Briefly, 12μL 0.1M ferric chloride was mixed with 2 mL of cell supernatant. After incubating at room temperature for 15 minutes, the precipitate was removed by centrifugation at 20000 × g for 15 min at 4 °C. The supernatant was added to an SPE plate (Waters, Oasis HLB) and washed with 0.5 mL 5% methanol, and then eluted with 0.5 mL 100% methanol to obtain the siderophores. LC/MS analysis was performed using 5μL aliquots. |
Combined analysis:
Analysis ID | AN003482 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 1290 Infinity |
Column | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Agilent 6560 Ion Mobility |
Ion Mode | POSITIVE |
Units | peak area |
Chromatography:
Chromatography ID: | CH002571 |
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: | MS003243 |
Analysis ID: | AN003482 |
Instrument Name: | Agilent 6560 Ion Mobility |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | Agilent MassHunter Workstation Data Acquisition Agilent MassHunter QualitativeAnalysis B.07.00 Agilent MassHunter Quantitative Analysis (for QTOF) |
Ion Mode: | POSITIVE |