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MB Sample ID: SA183138
Local Sample ID: | 20210306-4-30-1 |
Subject ID: | SU002016 |
Subject Type: | Bacteria |
Subject Species: | Escherichia coli |
Taxonomy ID: | 562 |
Species Group: | Bacteria |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002016 |
Subject Type: | Bacteria |
Subject Species: | Escherichia coli |
Taxonomy ID: | 562 |
Species Group: | Bacteria |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
20210306-4-30-1 | SA183138 | FL022306 | 0.15% | Concentration of MgSO4(w/v) |
Collection:
Collection ID: | CO002009 |
Collection Summary: | After 72 hours of culture, the planktonic cells were isolated, and the biofilms were washed with PBS solvent for three times. Then, planktonic cells and biofilm solutions were centrifuged at 2000 rpm and 4°C for 15 minutes, to collect the strain pellets. |
Sample Type: | Biofilm |
Treatment:
Treatment ID: | TR002028 |
Treatment Summary: | Colony-forming antigen (CFA) medium (1% casamino acids, 0.15% yeast extract, 0.005% magnesium sulfate, and 0.0005% manganese chloride) was used to form biofilms. 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 CFA medium at a ratio of 1:1000 and the cultures were incubated for another 72 h at 30°C to form mature biofilms. To test the influence of MgSO4 on biofilm formation, the purified compound of MgSO4 was added to the CFA medium at different concentrations for the preparation of treated culture medium; The left cultural procedures were kept as the same as the protocol listed above. |
Sample Preparation:
Sampleprep ID: | SP002022 |
Sampleprep Summary: | Each pellet sample was mixed with 2 mL of 80% ice-cold methanol, and the mixed solution was homogenized on ice for 2 minutes and repeated the same procedure for three times. Next, the collected samples were centrifuged at 12000 rpm and 4°C for 15 minutes, to collect the supernatants. The supernatants were further mixed with 800 μL of ice-cold acetonitrile for 20 minutes before further centrifuge at 12000 rpm and 4°C for 15 minutes. At last, the isolated supernatants were gone through a 0.22 μm filter membrane (nylon) before the lyophilization. The lyophilized samples were stored at -80°C for metabolomics analysis. |
Combined analysis:
Analysis ID | AN003151 | AN003152 |
---|---|---|
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 | NEGATIVE | POSITIVE |
Units | - | - |
Chromatography:
Chromatography ID: | CH002331 |
Chromatography Summary: | The chromatographic separation of biological samples was implemented on a Waters ACQUITY UPLC HSS T3 column (2.1×100 mm, 1.8 μm) with an optimized gradient-elution program (mobile phase A and B were 0.1% formic acid in water and acetonitrile (v/v)): 0-2 minutes, 98% A; 2-10 minutes, 98%-65% A; 10-12 minutes, 65%-20% A; 12-14 minutes, 20%-2% A; 14-30 minutes, 2% A. The column temperature was at 40°C and the flow rate was set at 0.3 mL/ minutes. |
Instrument Name: | Agilent 1290 Infinity |
Column Name: | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
Column Temperature: | 40 |
Flow Gradient: | 0-2 minutes, 98% A; 2-10 minutes, 98%-65% A; 10-12 minutes, 65%-20% A; 12-14 minutes, 20%-2% A; 14-30 minutes, 2% A |
Flow Rate: | 0.3 mL/ min |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100% acetonitrile; 0.1% formic acid |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS002931 |
Analysis ID: | AN003151 |
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 |
MS ID: | MS002932 |
Analysis ID: | AN003152 |
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 |