Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA331560

Local Sample ID:	Blank_1
Subject ID:	SU003168
Subject Type:	Bacteria
Subject Species:	Staphylococcus aureus
Taxonomy ID:	1280
Genotype Strain:	ATCC 43300
Age Or Age Range:	NA
Weight Or Weight Range:	NA
Height Or Height Range:	NA
Gender:	Not applicable

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Sample Preparation:

Sampleprep ID:	SP003174
Sampleprep Summary:	The bacterial pellets were washed twice in 1 mL of 0.9% saline and then centrifuged at 3,220 g and 4°C for 5 min to remove residual extracellular metabolites and medium components. The washed pellets were resuspended in a cold extraction solvent (chloroform-methanol-water at 1:3:1, vol/vol) containing 1 µM each of the internal standards 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), piperazine-N, N-bis (2-ethanesulfonic acid) (PIPES), and Tris. The samples were then frozen in liquid nitrogen, thawed on ice, and vortexed to release the intracellular metabolites (3 times). Next, the samples were transferred to 1.5-mL Eppendorf tubes and centrifuged at 14,000 g at 4°C for 10 min to remove any particulate matter. Finally, 200 µL of the supernatant was transferred into injection vials for liquid chromatography-mass spectrometry (LC-MS) analysis. An equal volume of each sample was combined and used as a quality control (QC) sample

Sampleprep ID:

SP003174

Sampleprep Summary:

The bacterial pellets were washed twice in 1 mL of 0.9% saline and then centrifuged at 3,220 g and 4°C for 5 min to remove residual extracellular metabolites and medium components. The washed pellets were resuspended in a cold extraction solvent (chloroform-methanol-water at 1:3:1, vol/vol) containing 1 µM each of the internal standards 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), piperazine-N, N-bis (2-ethanesulfonic acid) (PIPES), and Tris. The samples were then frozen in liquid nitrogen, thawed on ice, and vortexed to release the intracellular metabolites (3 times). Next, the samples were transferred to 1.5-mL Eppendorf tubes and centrifuged at 14,000 g at 4°C for 10 min to remove any particulate matter. Finally, 200 µL of the supernatant was transferred into injection vials for liquid chromatography-mass spectrometry (LC-MS) analysis. An equal volume of each sample was combined and used as a quality control (QC) sample