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MB Sample ID: SA166362
| Local Sample ID: | snf3_H5_C |
| Subject ID: | SU001863 |
| Subject Type: | Yeast |
| Subject Species: | Saccharomyces cerevisiae |
| Taxonomy ID: | 4932 |
| Gender: | Not applicable |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001863 |
| Subject Type: | Yeast |
| Subject Species: | Saccharomyces cerevisiae |
| Taxonomy ID: | 4932 |
| Gender: | Not applicable |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| snf3_H5_C | SA166362 | FL019817 | snf3 | Genotype |
| snf3_H5_C | SA166362 | FL019817 | H | Condition |
Collection:
| Collection ID: | CO001856 |
| Collection Summary: | For metabolomics, each replicate consisting of 3 mL of cell culture was mixed with 45 mL cold pure methanol on dry ice. After 5 min, cells were centrifuged in a precooled rotor (-80 °C). After discarding the supernatant, cell pellets were immediately stored at -80 °C. A small aliquot of each sample was saved to manually determine cell density with a hemocytometer. |
| Sample Type: | Yeast cells |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001876 |
| Treatment Summary: | No treatment in this study. |
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℃ |
Combined analysis:
| Analysis ID | AN002897 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Vanquish |
| Column | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH002148 |
| Chromatography Summary: | None |
| Chromatography Comments: | Time(min) Flow Rate %A %B Curve 1. 0 0.4 99.0 1.0 5 2. 1.00 0.4 99.0 1.0 5 3. 16.00 0.4 1.0 99.0 5 4. 19.00 0.4 1.0 99.0 5 5. 19.50 0.4 99.0 1.0 5 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
| Column Pressure: | 6000-10000 |
| Column Temperature: | 8 |
| Flow Rate: | 0.4 mL/min |
| Injection Temperature: | 8 |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% methanol; 0.1% formic acid |
| Weak Wash Solvent Name: | 10:90 Methanol:Water with 0.1% FA solution |
| Strong Wash Solvent Name: | 75:25 2-Propanol: Water with 0.1% FA solution |
| Randomization Order: | Yes |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002689 |
| Analysis ID: | AN002897 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | NA |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 275 °C |
| Capillary Voltage: | 3.5 KV |
| Collision Energy: | 10-35, ramp |
| Collision Gas: | N2 |
| Dry Gas Flow: | 45 |
| Dry Gas Temp: | 325°C |
| Fragmentation Method: | CID |
| Desolvation Gas Flow: | 45 |
