Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA077641

Local Sample ID:	33
Subject ID:	SU001174
Subject Type:	NMR based metabolomics of microbes
Subject Species:	Nesterenkonia flava
Taxonomy ID:	469799

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Subject:

Subject ID:	SU001174
Subject Type:	NMR based metabolomics of microbes
Subject Species:	Nesterenkonia flava
Taxonomy ID:	469799

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
33	SA077641	FL011808	1K00607	Strain
33	SA077641	FL011808	Marine	Strain source
33	SA077641	FL011808	A6	Medium

Collection:

Collection ID:	CO001168
Collection Summary:	The broth cultures (50 mL) were harvested via centrifugation at 7000 g for 10 min under 4 °C. The pellet was quenched using 10 mL of 60% cold methanol (−80 °C) containing 0.85% (wt./vol.) of NaCl for 30 min. The quenched cell pellets were re-suspended in 10 mL of cold PBS and were washed for 3 times.
Sample Type:	Cultured cells

Treatment:

Treatment ID:	TR001188
Treatment Summary:	For each cell pellet sample, 5 mL of the mixture of methanol/water (1:0.9, v/v) was added and ultrasonicated for 25 min to break cells bathing in ice. The mixture was added with 5 mL of cold chloroform, vortexed and subjected to 10 min of ultrasonic extraction under the bath of ice. The mixture was then centrifuged at 9500 g for 8 min. The upper layer phase containing methanol and water was taken out and evaporated in the fume cupboard to remove methanol. The remaining water solution was freezed under −80 °C, and then freeze-dried to afford dry samples, which were stored under −80 °C before analysis.

Treatment ID:

TR001188

Treatment Summary:

For each cell pellet sample, 5 mL of the mixture of methanol/water (1:0.9, v/v) was added and ultrasonicated for 25 min to break cells bathing in ice. The mixture was added with 5 mL of cold chloroform, vortexed and subjected to 10 min of ultrasonic extraction under the bath of ice. The mixture was then centrifuged at 9500 g for 8 min. The upper layer phase containing methanol and water was taken out and evaporated in the fume cupboard to remove methanol. The remaining water solution was freezed under −80 °C, and then freeze-dried to afford dry samples, which were stored under −80 °C before analysis.

Sample Preparation:

Sampleprep ID:	SP001181
Sampleprep Summary:	The intracellular extract was re-suspended in 550 µL of phosphate buffer containing 1.5 M KH2PO4, 0.1% sodium 3-(trimethylsilyl) propionate-2,2,3,3-d4 (TSP), and 10% D2O. Subsequently, all the samples were vortexed and centrifuged at 12000 g for 15 min at 4°C to remove any insoluble components. The collected supernatants (500 μL) were transferred to 5 mm NMR tubes for analysis.

Analysis:

MB Sample ID:	SA077641
Analysis ID:	AN001828
Analysis Type:	NMR
Num Factors:	12

NMR:

NMR ID:	NM000141
Analysis ID:	AN001828
Instrument Name:	Bruker Avance III
Instrument Type:	FT-NMR
NMR Experiment Type:	1D 1H
Spectrometer Frequency:	850 MHz