Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA169193

Local Sample ID:	28
Subject ID:	SU001900
Subject Type:	Mammal
Subject Species:	Equus caballus
Taxonomy ID:	9796

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Sample Preparation:

Sampleprep ID:	SP001906
Sampleprep Summary:	Five-hundred mg of feces was aliquoted into a 2ml tube, lyophilized overnight and vortexed with a 5mm stainless steel bead (Quiagen, Germantown, MD ) for 5 minutes. Samples were then extracted using a methanol:chloroform:water-based extraction method. Briefly 800 uL ice cold methanol:chloroform (1:1, v:v) was added to samples in a bead-based lysis tube (Bertin, Rockville, MD). Samples were homogenized for 30 seconds on a Precyllys 24 (Bertin, Rockville, MD) at a speed of 6000. The supernatant was collected and samples were homogenized a second time with 800 uL ice methanol:chloroform. 600 uL ice cold water was added to the combined extract, vortexed and centrifuged to separate the phases. The upper aqueous layer was passed through a 0.2 um nylon filter (Merck Millipore, Burlington, MA). 500 uL of the filtered aqueous phase was then passed through a 3 kDa cutoff column (Thermo Scientific, Waltham, MA) and the flow through was collected for analysis.

Sampleprep ID:

SP001906

Sampleprep Summary:

Five-hundred mg of feces was aliquoted into a 2ml tube, lyophilized overnight and vortexed with a 5mm stainless steel bead (Quiagen, Germantown, MD ) for 5 minutes. Samples were then extracted using a methanol:chloroform:water-based extraction method. Briefly 800 uL ice cold methanol:chloroform (1:1, v:v) was added to samples in a bead-based lysis tube (Bertin, Rockville, MD). Samples were homogenized for 30 seconds on a Precyllys 24 (Bertin, Rockville, MD) at a speed of 6000. The supernatant was collected and samples were homogenized a second time with 800 uL ice methanol:chloroform. 600 uL ice cold water was added to the combined extract, vortexed and centrifuged to separate the phases. The upper aqueous layer was passed through a 0.2 um nylon filter (Merck Millipore, Burlington, MA). 500 uL of the filtered aqueous phase was then passed through a 3 kDa cutoff column (Thermo Scientific, Waltham, MA) and the flow through was collected for analysis.