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MB Sample ID: SA076769
| Local Sample ID: | 12-Late Exponential Phase |
| Subject ID: | SU001169 |
| Subject Type: | Bacteria |
| Subject Species: | Acidithiobacillus ferrooxidans ATCC 23270 |
| Taxonomy ID: | 243159 |
| Genotype Strain: | ATCC 23270 |
| Cell Biosource Or Supplier: | American Type Culture Collection |
| Cell Strain Details: | ATCC 23270 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001169 |
| Subject Type: | Bacteria |
| Subject Species: | Acidithiobacillus ferrooxidans ATCC 23270 |
| Taxonomy ID: | 243159 |
| Genotype Strain: | ATCC 23270 |
| Cell Biosource Or Supplier: | American Type Culture Collection |
| Cell Strain Details: | ATCC 23270 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 12-Late Exponential Phase | SA076769 | FL011741 | 0.85 | Optic Density(OD) |
Collection:
| Collection ID: | CO001163 |
| Collection Summary: | Replicate cultures of Acidithiobacillus ferrooxidans (American Type Culture Collection strain 23270) were grown in 1 L of Tuovinen and Kelly (1973) liquid medium ([FeSO4·7H2O] = 33.4 g/L; 0.3 % (w/v) (NH4)2SO4; MgSO4·7H2O; K2HPO4, pH 2.5). Cells were incubated on a gyratory shaker at 180 rpm and 30 °C. Cells harvested for the metabolomic comparison of two points in the growth curve were in late exponential growth phase (OD425 = 0.85) or in early exponential growth phase (OD425 = 0.65). The cells were harvested via centrifugation at 14,000 x g for 5 min at 4 °C then transferred into a 1.5 mL centrifuge tube. |
| Sample Type: | Bacterial cells |
Treatment:
| Treatment ID: | TR001183 |
| Treatment Summary: | After the cells were concentrated into a 1.5ml centrifuge tube, the cellular metabolism was quenched using 40ul of 60% MeOH and 40% H2O (v/v) with 0.85% (w/v) of Ammonium Formate, adjusted to pH 2.5 and cooled to -20 °C. The quenched cells were centrifuged for 5 minutes at 2,319 x g at 4 °C. The cells were then placed on dry ice, the cell supernatant was removed and an isopropanol:methanol:water (IMW) (3:3:2) extraction solution that was cooled to -20 °C was added. These solutions were then vigorously vortexed and then centrifuged at 15,682 x g for 10 min. The extraction liquid was then collected and placed at -80 °C until further analysis. |
Sample Preparation:
| Sampleprep ID: | SP001176 |
| Sampleprep Summary: | Immediately prior to analysis the extraction solution was evaporated to dryness and reconstituted in 80:20 Acetonitrile:Water. The samples were normalized based on Gcdw. |
Combined analysis:
| Analysis ID | AN001821 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 |
| Column | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Bruker MicrOTOF II |
| Ion Mode | POSITIVE |
| Units | m/z-retention time features |
Chromatography:
| Chromatography ID: | CH001290 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| Flow Gradient: | (t= 0-1 minute) 3% A, 97% B; (t= 24 minute) 40% A, 60% B; (t= 28 minute) 40% A, 60% B (t= 33 minutes) 3% A, 97% B; (t= 34minutes) 3% A, 97% B. HPLC flow rate 0.4 mL/min. |
| Flow Rate: | 0.4ml/min |
| Solvent A: | 100% water; 20 mM ammonium formate |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS001684 |
| Analysis ID: | AN001821 |
| Instrument Name: | Bruker MicrOTOF II |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | None |
| Ion Mode: | POSITIVE |
