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MB Sample ID: SA166243
| Local Sample ID: | WT_L1_B |
| Subject ID: | SU001863 |
| Subject Type: | Yeast |
| Subject Species: | Saccharomyces cerevisiae |
| Taxonomy ID: | 4932 |
| Gender: | Not applicable |
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Sample Preparation:
| Sampleprep ID: | SP001869 |
| Sampleprep Summary: | Frozen cell pellets were resuspended with extraction reagent (8:2 methanol-water solution) to 3x108 cell/mL and then transferred into 2 mL ceramic bead MagNalyser tubes. Blank samples were prepared by adding 1300 μL of extraction reagent with no cells to a MagNalyser tube with ceramic beads. Tubes were subjected to homogenization, with Bead Ruptor Elite Bead Mill Homogenizer (OMNI International) at 6.0 m/s for 40 sec in 2 cycles at room temperature. This step was repeated twice. All samples were then centrifuged at 16,000 xg for 10 min at 4 °C. 500 μL of the supernatant was transferred into low-bind 1.7 mL microfuge tubes. Total pools were made by combining an additional 65 μL of the supernatant from each sample and then aliquoting this mixture into low-bind 1.7 mL tubes at a volume of 500 μL. The remaining supernatant was stored at -80 °C for repeat experiments if necessary. For all experimental samples, pooled samples and blanks were dried using a speedvac vacuum concentrator overnight. Dried samples were stored at -80 °C. Before LC-MS analysis, 100 μL of reconstitution buffer (95:5 water:methanol with 500 ng/mL tryptophan d-5) was added to each dried sample. All tubes were vortexed at 5000 rpm for 10 min and then centrifuged at room temperature at 16,000 xg for 4 min. Supernatant was transferred into autosampler vials for LC-MS. |
| Processing Storage Conditions: | 4℃ |
| Extract Storage: | -80℃ |
