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MB Sample ID: SA113898

Local Sample ID:WT at 3hr minR+Leu Sample3
Subject ID:SU001475
Subject Type:Yeast
Subject Species:Saccharomyces cerevisiae
Taxonomy ID:4932
Genotype Strain:BY4743
Gender:Not applicable

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Subject:

Subject ID:SU001475
Subject Type:Yeast
Subject Species:Saccharomyces cerevisiae
Taxonomy ID:4932
Genotype Strain:BY4743
Gender:Not applicable

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
WT at 3hr minR+Leu Sample3SA113898FL014359wild-typeGenotype
WT at 3hr minR+Leu Sample3SA113898FL014359minR+LeuTreatment
WT at 3hr minR+Leu Sample3SA113898FL014359180 minTime

Collection:

Collection ID:CO001470
Collection Summary:Metabolomics data were generated by growing the appropriate yeast strains in synthetic minimal media (S-min) supplemented with 2% glucose until they reached OD>600 between 0.6-0.8. Cells were then transferred to S-min media containing 2% raffinose and harvested after 0, 15, 30, 60, and 180 minutes (n=6/time-point/strain; except in one 3 hr wild-type sample, where n=5).
Sample Type:Yeast cells
Collection Location:Rutter Lab, University of Utah

Treatment:

Treatment ID:TR001490
Treatment Summary:Metabolomics data were generated by growing the appropriate yeast strains in synthetic minimal media (S-min) supplemented with 2% glucose until they reached OD>600 between 0.6-0.8. Cells were then transferred to S-min media containing 2% raffinose and harvested after 0, 15, 30, 60, and 180 minutes (n=6/time-point/strain; except in one 3 hr wild-type sample, where n=5).

Sample Preparation:

Sampleprep ID:SP001483
Sampleprep Summary:To each sample was added boiling 75% ethanol (EtOH) solution containing the internal standard d4-succinic acid (Sigma 293075). Boiling samples were vortexed and incubated at 90°C for 5 min. Samples were then incubated at -20 ˚C for 1 hr. After incubation the samples were centrifuged at 5,000 x g for 10 minutes at 4˚C. The supernatant was then transferred from each sample tube into a labeled, fresh 13x100mm glass culture tube. A second standard was then added (d27-myristic acid CDN Isotopes: D-1711). Pooled quality control samples were made by removing a fraction of collected supernatant from each sample and process blanks were made using only extraction solvent and no cell culture. The samples were then dried en vacuo. This process was completed in three separate batches.

Combined analysis:

Analysis ID AN002343
Analysis type MS
Chromatography type GC
Chromatography system Agilent 5977b
Column Phenomenex Zebron AB-5HT (5m)
MS Type EI
MS instrument type HES-MSD
MS instrument name Agilent 5977
Ion Mode POSITIVE
Units AUC (unitless)

Chromatography:

Chromatography ID:CH001716
Chromatography Summary:All GC-MS analysis was performed with an Agilent 5977b GC-MS MSD-HES and an Agilent 7693A automatic liquid sampler. Dried samples were suspended in 40 µL of a 40 mg/mL O-methoxylamine hydrochloride (MOX) (MP Bio #155405) in dry pyridine (EMD Millipore #PX2012-7) and incubated for one hour at 37 °C in a sand bath. 25 µL of this solution was added to auto sampler vials. 60 µL of N-methyl-N-trimethylsilyltrifluoracetamide (MSTFA with 1%TMCS, Thermo #TS48913) was added automatically via the auto sampler and incubated for 30 minutes at 37 °C. After incubation, samples were vortexed and 1 µL of the prepared sample was injected into the gas chromatograph inlet in the split mode with the inlet temperature held at 250°C. A 10:1 split ratio was used for analysis of the majority of metabolites. The gas chromatograph had an initial temperature of 60°C for one minute followed by a 10°C/min ramp to 325°C and a hold time of 5 minutes. A 30-meter Phenomenex Zebron AB-5HT with 5m inert Guardian capillary column was employed for chromatographic separation. Helium was used as the carrier gas at a rate of 1 mL/min.
Instrument Name:Agilent 5977b
Column Name:Phenomenex Zebron AB-5HT (5m)
Chromatography Type:GC

MS:

MS ID:MS002185
Analysis ID:AN002343
Instrument Name:Agilent 5977
Instrument Type:HES-MSD
MS Type:EI
MS Comments:All GC-MS analysis was performed with an Agilent 5977b GC-MS MSD-HES and an Agilent 7693A automatic liquid sampler. Dried samples were suspended in 40 µL of a 40 mg/mL O-methoxylamine hydrochloride (MOX) (MP Bio #155405) in dry pyridine (EMD Millipore #PX2012-7) and incubated for one hour at 37 °C in a sand bath. 25 µL of this solution was added to auto sampler vials. 60 µL of N-methyl-N-trimethylsilyltrifluoracetamide (MSTFA with 1%TMCS, Thermo #TS48913) was added automatically via the auto sampler and incubated for 30 minutes at 37 °C. After incubation, samples were vortexed and 1 µL of the prepared sample was injected into the gas chromatograph inlet in the split mode with the inlet temperature held at 250°C. A 10:1 split ratio was used for analysis of the majority of metabolites. For those metabolites that saturated the instrument at the 10:1 split concentration, a split of 50:1 was used for analysis. The gas chromatograph had an initial temperature of 60°C for one minute followed by a 10°C/min ramp to 325°C and a hold time of 5 minutes. A 30-meter Phenomenex Zebron AB-5HT with 5m inert Guardian capillary column was employed for chromatographic separation. Helium was used as the carrier gas at a rate of 1 mL/min. Data was collected using MassHunter software (Agilent). Metabolites were identified and their peak area was recorded using MassHunter Quant. This data was transferred to an Excel spread sheet (Microsoft, Redmond WA). Metabolite identity was established using a combination of an in-house metabolite library developed using pure purchased standards, the NIST library and the Fiehn library.
Ion Mode:POSITIVE
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