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MB Sample ID: SA094377
Local Sample ID: | WT8 |
Subject ID: | SU001379 |
Subject Type: | Bacteria |
Subject Species: | Salmonella enterica |
Taxonomy ID: | 28901 |
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Subject:
Subject ID: | SU001379 |
Subject Type: | Bacteria |
Subject Species: | Salmonella enterica |
Taxonomy ID: | 28901 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
WT8 | SA094377 | FL013485 | WT | Strain |
Collection:
Collection ID: | CO001374 |
Collection Summary: | Ten biologically independent cultures each of wild-type (DM9404) and ridA mutant (DM3480) strains were grown overnight in NB medium at 37 °C and used to inoculate (1% inoculum) 250 mL minimal glucose medium in 500 mL non-baffled flasks. Flasks were randomly arranged in an Innova®44 incubator and cultures were allowed to grow 16 h shaking at 180 RPM and 37 °C. Cultures were cooled on ice 5 min and then harvested by centrifugation at 7,000 x G for 10 min at 4 °C. The supernatant was decanted, pellets were resuspended in 10 mL ddH2O and transferred to sterile 15 mL conical tubes in which they were pelleted at 7,000 x G 10 min at 4 °C. Final supernatant was decanted and pellets were frozen in liquid nitrogen and stored at -80 °C prior to cell extractions. |
Sample Type: | Bacterial cells |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001394 |
Treatment Summary: | These samples did not undergo addition treatment. |
Sample Preparation:
Sampleprep ID: | SP001387 |
Sampleprep Summary: | Each frozen bacterial pellet (~1 g) was thawed on ice and homogenized 3 times. For homogenization, 2.6 g of 0.1 mm zirconium beads and 6 mL 4 C MeOH/H2O (80/20) solvent was added and samples were agitated 7 times (210 s total) at 1600 rpm in a FastPrep 96 (MPBIO). Samples were then centrifuged at 416 g and 4 C for 16 min and supernatant was transferred to a new 15 mL conical tube. Homogenization was carried out a second and third time using 4 mL MeOH/H2O with 4 homogenization cycles (150 s) and 2 mL MeOH/H2O with 3 homogenization cycles (150 s), respectively. Pooled supernatants from each sample were concentrated to dryness using a CentriVap Benchtop Vacuum Concentrator (Labconco). The extracts were reconstituted in 600 L of deuterated 100 mM sodium phosphate buer (pH 7.4) containing 1 mM of the internal standard DSS (d6 4,4-dimethyl-4-silapentane-1-sulfonic acid) and vortex mixed 2 min. Each sample was transferred into 5 mm SampleJet NMR tubes for NMR analysis. |
Analysis:
MB Sample ID: | SA094377 |
Analysis ID: | AN002174 |
Laboratory Name: | Complex Carbohydrate Research Center NMR Facility |
Analysis Type: | NMR |
Operator Name: | Adrien Le Guennec |
Num Factors: | 2 |
Num Metabolites: | 37 |
Units: | Area Under Curve |
NMR:
NMR ID: | NM000159 |
Analysis ID: | AN002174 |
Instrument Name: | Bruker Avance III HD |
Instrument Type: | FT-NMR |
NMR Experiment Type: | 1D-1H |
Spectrometer Frequency: | 800 MHz |