Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA235671

Local Sample ID:	JM SQ 4
Subject ID:	SU002437
Subject Type:	Bacteria
Subject Species:	Escherichia coli
Taxonomy ID:	562
Genotype Strain:	BW25113

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Sample Preparation:

Sampleprep ID:	SP002443
Sampleprep Summary:	Cell pellets were resuspended in chilled extraction solution (500 µL) comprised of 3:1 MeOH: H2O and (13C5, 15N)valine (1 mM, 0.5 µL) and (13C6)sorbitol (1 mM, 0.5 µL) as internal standards. The suspensions were subjected to freeze-thaw cycles to facilitate lysis of the cells (30 s in liquid N2, 30 s in a dry ice/EtOH bath for 10 cycles), and shaken (9000 rpm, 10 min, 2°C). The samples were then centrifuged (12700 rpm, 5 min, 1°C). The cell lysate supernatant was transferred into glass inserts and dried. All samples were washed with MeOH (50 µL). All samples were methoximated with methoxylamine hydrochloride solution (30 mg/mL in pyridine, 20 µL) for 2 h at 37°C, followed by trimethylsilyation in BSTFA + 1% TMCS (20 µL) for 1 h at 37°C with continuous mixing. Samples were incubated at rt for 1 h prior to GC-MS analysis.

Sampleprep ID:

SP002443

Sampleprep Summary:

Cell pellets were resuspended in chilled extraction solution (500 µL) comprised of 3:1 MeOH: H2O and (13C5, 15N)valine (1 mM, 0.5 µL) and (13C6)sorbitol (1 mM, 0.5 µL) as internal standards. The suspensions were subjected to freeze-thaw cycles to facilitate lysis of the cells (30 s in liquid N2, 30 s in a dry ice/EtOH bath for 10 cycles), and shaken (9000 rpm, 10 min, 2°C). The samples were then centrifuged (12700 rpm, 5 min, 1°C). The cell lysate supernatant was transferred into glass inserts and dried. All samples were washed with MeOH (50 µL). All samples were methoximated with methoxylamine hydrochloride solution (30 mg/mL in pyridine, 20 µL) for 2 h at 37°C, followed by trimethylsilyation in BSTFA + 1% TMCS (20 µL) for 1 h at 37°C with continuous mixing. Samples were incubated at rt for 1 h prior to GC-MS analysis.