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MB Sample ID: SA315462
| Local Sample ID: | D1_2 |
| Subject ID: | SU003004 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Age Or Age Range: | 15-20 week old |
| Gender: | Male |
| Species Group: | C57BL/6J |
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Sample Preparation:
| Sampleprep ID: | SP003010 |
| Sampleprep Summary: | Frozen samples were stored at -80°C until analyzed by LC-MS-based metabolomics in the Vanderbilt Center for Innovative Technology (CIT). Isotopically labeled phenylalanine-D8 and biotin-D2 were added to 200 μL of culture supernatant per sample, and protein was precipitated by addition of 800 μL of ice-cold methanol followed by overnight incubation at −80°C. Precipitated proteins were pelleted by centrifugation (15,000 rpm, 15 min), and extracted metabolites were dried down in vacuo and stored at −80°C. Individual samples were reconstituted in 100 μL of reconstitution buffer (acetonitrile/water, 90:10, vol/vol) containing tryptophan-D3 and inosine-4N15. Equal volumes of individual samples were pooled to create a quality control (QC) pooled sample used for column conditioning, retention time alignment, to assess instrument reproducibility, and for batch acceptance. |
| Processing Storage Conditions: | -80℃ |
| Extract Storage: | -80℃ |
