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MB Sample ID: SA255564
| Local Sample ID: | met6KO_15h_2 |
| Subject ID: | SU002641 |
| Subject Type: | Yeast |
| Subject Species: | Saccharomyces cerevisiae |
| Taxonomy ID: | 4932 |
| Genotype Strain: | CEN.P.K |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002641 |
| Subject Type: | Yeast |
| Subject Species: | Saccharomyces cerevisiae |
| Taxonomy ID: | 4932 |
| Genotype Strain: | CEN.P.K |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| met6KO_15h_2 | SA255564 | FL032714 | 15h | Sample Type |
Collection:
| Collection ID: | CO002634 |
| Collection Summary: | Care was taken to quench cells quickly in -40℃ and maintain metabolites in acid to minimize oxidation |
| Sample Type: | Yeast cells |
Treatment:
| Treatment ID: | TR002653 |
| Treatment Summary: | Mutant MET6KO cells were cultured in SD medium with the supplements of Met and growed to log phase, then cells were washed with normal SD medium and re-cultured in SD medium |
Sample Preparation:
| Sampleprep ID: | SP002647 |
| Sampleprep Summary: | Briefly, equal OD units of cells were rapidly quenched to stop metabolism by addition into 4 volumes of quenching buffer containing 60% methanol and 10 mM Tricine, pH 7.4 that was pre-cooled to -40°C. 5 min after holding at -40°C, cells were spun at 5,000 g for 3 min at 0°C, washed with the same buffer, and then resuspended in 1 mL extraction buffer containing 75% ethanol and 0.5 mM Tricine, pH 7.4. Intracellular metabolites were extracted by incubating at 75°C for 3 min, followed by chilling on ice for 5 min. Samples were spun at 20,000 g for 1 min to pellet cell debris, and 0.9 mL of the supernatant was transferred to a new tube. After a second spin at 20,000 g for 10 min, 0.8 mL of the supernatant was transferred to a new tube. Metabolites in the extraction buffer were dried using a vacuum concentrator system (Labconco) and stored at -80°C until analysis. Dried metabolite extracts were resuspended in 60% acetonitrile for injection. |
Combined analysis:
| Analysis ID | AN004186 | AN004187 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Shimadzu 20AD | Shimadzu 20AD |
| Column | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| MS Type | ESI | ESI |
| MS instrument type | QTRAP | QTRAP |
| MS instrument name | ABI Sciex 6500+ | ABI Sciex 6500+ |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Peak intensity | Peak intensity |
Chromatography:
| Chromatography ID: | CH003102 |
| Chromatography Summary: | Low pH polar |
| Instrument Name: | Shimadzu 20AD |
| Column Name: | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0.01 min 80% B, 20 min 20% B, 20.5 min 80% B, 34 min 80% B. |
| Flow Rate: | 0.15ml/min |
| Solvent A: | 20 mM ammonium carbonate and 0.1% (v/v) ammonium hydroxide |
| Solvent B: | acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003933 |
| Analysis ID: | AN004186 |
| Instrument Name: | ABI Sciex 6500+ |
| Instrument Type: | QTRAP |
| MS Type: | ESI |
| MS Comments: | SCIEX OS |
| Ion Mode: | POSITIVE |
| MS ID: | MS003934 |
| Analysis ID: | AN004187 |
| Instrument Name: | ABI Sciex 6500+ |
| Instrument Type: | QTRAP |
| MS Type: | ESI |
| MS Comments: | SCIEX OS |
| Ion Mode: | NEGATIVE |
