Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA168223

Local Sample ID:	GATp10GLN-1-pos-PRM
Subject ID:	SU001888
Subject Type:	Plant
Subject Species:	Arabidopsis thaliana
Taxonomy ID:	3702
Genotype Strain:	Col-0
Age Or Age Range:	10 day old seedlings
Gender:	Not applicable

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Sample Preparation:

Sampleprep ID:	SP001894
Sampleprep Summary:	Fifty mg of root tissue was excised from 10 day old seedlings of WT or gat1_2.1 grown under conditions described above, collected in 2 mL Eppendorf tubes and flash frozen in liquid N2. Frozen tissue was homogenized using a tissue lyser and metabolites were isolated using 1 mL of methanol:water (4:1) with incubation in an ultra-sonication bath for 20 min followed by shaking for 30 min at 4 °C. The mixture was centrifuged at 12,000 × g for 10 min at 4 °C and 700 µl of the supernatant was transferred into fresh tubes and evaporated to dryness using a Vacufuge at ambient temperature. The residue was re-dissolved in 500 µl of 1:1 methanol:water and the samples were filtered using a 0.2 µm PTFE microfuge filter (Cytiva Whatman). Five µl of 1 µg/mL 13C6 Phe was added to the samples for monitoring the quality of LC-MS runs.

Sampleprep ID:

SP001894

Sampleprep Summary:

Fifty mg of root tissue was excised from 10 day old seedlings of WT or gat1_2.1 grown under conditions described above, collected in 2 mL Eppendorf tubes and flash frozen in liquid N2. Frozen tissue was homogenized using a tissue lyser and metabolites were isolated using 1 mL of methanol:water (4:1) with incubation in an ultra-sonication bath for 20 min followed by shaking for 30 min at 4 °C. The mixture was centrifuged at 12,000 × g for 10 min at 4 °C and 700 µl of the supernatant was transferred into fresh tubes and evaporated to dryness using a Vacufuge at ambient temperature. The residue was re-dissolved in 500 µl of 1:1 methanol:water and the samples were filtered using a 0.2 µm PTFE microfuge filter (Cytiva Whatman). Five µl of 1 µg/mL 13C6 Phe was added to the samples for monitoring the quality of LC-MS runs.