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MB Sample ID: SA081514
Local Sample ID: | SrrAB_2 |
Subject ID: | SU001240 |
Subject Type: | Bacteria |
Subject Species: | Staphylococcus aureus |
Taxonomy ID: | 1280 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001240 |
Subject Type: | Bacteria |
Subject Species: | Staphylococcus aureus |
Taxonomy ID: | 1280 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
SrrAB_2 | SA081514 | FL012427 | SrrAB deletion mutant | Genotype |
Collection:
Collection ID: | CO001234 |
Collection Summary: | Aerobic growth for all strains was performed on four biological replicates for each cell group. Overnight cultures diluted 1:1000 were used to inoculate 25 mL of fresh TSB in a 250 mL flask with 220 rpm agitation at 37° C. Aliquots of 10 mL were collected at 48 hrs , centrifuged at 5000 rpm for 5 minutes, then rinsed once with 1 mL of 1X PBS, and centrifuged at 5000 rpm for 5 minutes. The supernatant was discarded, and cell pellets were frozen at -80 °C until further use. |
Sample Type: | Bacterial cells |
Treatment:
Treatment ID: | TR001255 |
Treatment Summary: | deletion mutants were compared to the wild-type after 48 hours of growth |
Sample Preparation:
Sampleprep ID: | SP001248 |
Sampleprep Summary: | Frozen cell pellets were resuspended in 1 mL of a 2:1 methanol/chloroform mixture and transferred to FastPrep lysis B matrix tubes (MP Biomedicals). Cells were lysed using the FastPrep-24 5G instrument and designated S. aureus settings (2 cycles at a speed of 6.0 m/s for 40 s)]. 300 μL of each layer of a 1:1 aqueous chloroform solution was added to each cell lysate. The tubes were vortexed, placed at -20 °C for 20 min, and centrifuged at 14,000 g for 10 minutes. 800 μL of the aqueous phase was transferred to microfuge tubes and placed in a Speedvac (no heat, manual run, volatile solvent) to dry overnight. Samples were resuspended in 600 μL of NMR buffer [0.25 mM 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS), 0.4 mM imidazole, 25 mM phosphate buffer, 90% H2O/10% D2O] and transferred to 5 mm Bruker NMR tubes. |
Analysis:
MB Sample ID: | SA081514 |
Analysis ID: | AN001949 |
Analysis Type: | NMR |
Operator Name: | Brian Tripet |
Num Factors: | 4 |
Num Metabolites: | 38 |
Units: | Attomoles/CFU |
NMR:
NMR ID: | NM000148 |
Analysis ID: | AN001949 |
Instrument Name: | Bruker 600-MHz AVANCE III solution NMR spectrometer |
Instrument Type: | FT-NMR |
NMR Experiment Type: | 1D-1H |
NMR Comments: | equipped with an automatic SampleJet sample loading system, as well as a 5 mm triple resonance (1H, 15N, 13C) liquid-helium-cooled TCI probe (cryoprobe) and Topspin software (Bruker version 3.2). |
Spectrometer Frequency: | 1H Larmor frequency |
NMR Probe: | cryoprobe |
NMR Solvent: | 10%D20 |
NMR Tube Size: | 5mm |
Shimming Method: | topshim |
Pulse Sequence: | zgesgp |
Water Suppression: | qfil |