Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA081517

Local Sample ID:	WT_1
Subject ID:	SU001240
Subject Type:	Bacteria
Subject Species:	Staphylococcus aureus
Taxonomy ID:	1280

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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
WT_1	SA081517	FL012428	wild type	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.

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:	SA081517
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