Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA308653

Local Sample ID:	tumor-veh-3-01
Subject ID:	SU002964
Subject Type:	Mammal
Subject Species:	Mus musculus
Genotype Strain:	NSG

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Sample Preparation:

Sampleprep ID:	SP002970
Sampleprep Summary:	After tumor dissection, a maximum of 30mg of tissue was weighed, snap frozen, and stored at -80C until metabolite extraction. To extract metabolites, weighed tumor tissue was added to a bead tube (Thermo Fisher Scientific) containing 1ml 80% methanol plus 10mM potassium trifluoromethanesulfonate (TMSO) internal standard on ice. Samples were homogenized for 1 minute at max speed on a bead homogenizer (Thermo Fisher Scientific). Bead tubes were spun at 17000g at 4C for 10 minutes. The supernatant was transferred to an Eppendorf tube and spun at 17000g at 4C for 10 minutes. A volume of extraction equivalent to 3mg of tumor tissue was transferred to an ABC vial (Thermo Fisher Scientific). All volumes were normalized to 500 µl with 80% methanol containing TMSO internal standard. 80% MeOH was evaporated from the ABC vials using the EZ-2Elite evaporator (Genevac) and samples were stored at -80°C until analysis. Dried metabolites were reconstituted in 100 µL of a 50% acetonitrile (ACN) 50% dH20 solution. Samples were vortexed and spun down for 10 minutes at 17,000g. 70 µL of the supernatant was then transferred to HPLC glass vials. 10 µL of these metabolite solutions were injected per analysis.

Sampleprep ID:

SP002970

Sampleprep Summary:

After tumor dissection, a maximum of 30mg of tissue was weighed, snap frozen, and stored at -80C until metabolite extraction. To extract metabolites, weighed tumor tissue was added to a bead tube (Thermo Fisher Scientific) containing 1ml 80% methanol plus 10mM potassium trifluoromethanesulfonate (TMSO) internal standard on ice. Samples were homogenized for 1 minute at max speed on a bead homogenizer (Thermo Fisher Scientific). Bead tubes were spun at 17000g at 4C for 10 minutes. The supernatant was transferred to an Eppendorf tube and spun at 17000g at 4C for 10 minutes. A volume of extraction equivalent to 3mg of tumor tissue was transferred to an ABC vial (Thermo Fisher Scientific). All volumes were normalized to 500 µl with 80% methanol containing TMSO internal standard. 80% MeOH was evaporated from the ABC vials using the EZ-2Elite evaporator (Genevac) and samples were stored at -80°C until analysis. Dried metabolites were reconstituted in 100 µL of a 50% acetonitrile (ACN) 50% dH20 solution. Samples were vortexed and spun down for 10 minutes at 17,000g. 70 µL of the supernatant was then transferred to HPLC glass vials. 10 µL of these metabolite solutions were injected per analysis.