Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA159028

Local Sample ID:	POS_WT_m1663_1
Subject ID:	SU001784
Subject Type:	Cultured cells
Subject Species:	Mus musculus
Taxonomy ID:	10090

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Sample Preparation:

Sampleprep ID:	SP001790
Sampleprep Summary:	Following aqueous extraction using cold methanol and water (v:v, 1:1), 1 ml of pre-chilled dichloromethane (DCM)/methanol (v:v, 3:1) was added to the organoid samples. Samples were bead-beaten for 40 seconds followed by five minutes of chilling on dry ice. This procedure was repeated three times before being centrifuged for 10 mins at 21,000 rcf at 4ºC. A total of 600 μl of supernatant from each sample was transferred to a glass vial. Another 200 μl of supernatant from each sample was pooled into a 15-ml Falcon tube to form a quantity control (QC) sample and split into several aliquots of 600 μl each. An extraction blank sample was included to control for any potential contaminant introduced throughout the extraction process. Samples were dried by evaporation over night at room temperature and stored at -40˚C until further analysis. The dried extracts were reconstituted in 100 μl of water/acetonitrile (ACN)/isopropanol (IPA), (v:v:v, 1:1:3,). The lipids were dissolved by vigorous vortexing for five minutes, followed by five minutes of sonication. This step was repeated three times to allow the dry extracts to thoroughly dissolve in the solvent. Samples were subsequently centrifuged at 21,000 rcf for 10 minutes at 4ºC and transferred to 150-μl glass inserts placed in glass vials (Waters).

Sampleprep ID:

SP001790

Sampleprep Summary:

Following aqueous extraction using cold methanol and water (v:v, 1:1), 1 ml of pre-chilled dichloromethane (DCM)/methanol (v:v, 3:1) was added to the organoid samples. Samples were bead-beaten for 40 seconds followed by five minutes of chilling on dry ice. This procedure was repeated three times before being centrifuged for 10 mins at 21,000 rcf at 4ºC. A total of 600 μl of supernatant from each sample was transferred to a glass vial. Another 200 μl of supernatant from each sample was pooled into a 15-ml Falcon tube to form a quantity control (QC) sample and split into several aliquots of 600 μl each. An extraction blank sample was included to control for any potential contaminant introduced throughout the extraction process. Samples were dried by evaporation over night at room temperature and stored at -40˚C until further analysis. The dried extracts were reconstituted in 100 μl of water/acetonitrile (ACN)/isopropanol (IPA), (v:v:v, 1:1:3,). The lipids were dissolved by vigorous vortexing for five minutes, followed by five minutes of sonication. This step was repeated three times to allow the dry extracts to thoroughly dissolve in the solvent. Samples were subsequently centrifuged at 21,000 rcf for 10 minutes at 4ºC and transferred to 150-μl glass inserts placed in glass vials (Waters).