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MB Sample ID: SA255574
Local Sample ID: | met6KO_6h_3 |
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_6h_3 | SA255574 | FL032717 | 6h | 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 |