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MB Sample ID: SA168246
Local Sample ID: | WT-GLN-1-neg-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. |