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MB Sample ID: SA204418
Local Sample ID: | fur-0-4S |
Subject ID: | SU002213 |
Subject Type: | Bacteria |
Subject Species: | Escherichia coli |
Taxonomy ID: | 562 |
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Subject:
Subject ID: | SU002213 |
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-0-4S | SA204418 | FL025108 | No iron supplementation | Treatment |
Collection:
Collection ID: | CO002206 |
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: | TR002225 |
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 siderophores. 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: | SP002219 |
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 | AN003481 |
---|---|
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: | CH002570 |
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: | MS003242 |
Analysis ID: | AN003481 |
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 |