Return to study ST000887 main page
MB Sample ID: SA076768
Local Sample ID: | 4-Early 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 |
---|---|---|---|---|
4-Early Exponential Phase | SA076768 | FL011740 | 0.65 | 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 |