Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA136789

Local Sample ID:	YGR023w-4
Subject ID:	SU001689
Subject Type:	Yeast
Subject Species:	Saccharomyces cerevisiae
Taxonomy ID:	4932
Genotype Strain:	BY4742, YGR023w
Cell Biosource Or Supplier:	Open Biosystems

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Subject:

Subject ID:	SU001689
Subject Type:	Yeast
Subject Species:	Saccharomyces cerevisiae
Taxonomy ID:	4932
Genotype Strain:	BY4742, YGR023w
Cell Biosource Or Supplier:	Open Biosystems

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
YGR023w-4	SA136789	FL017504	mtl1Δ	Strain

Collection:

Collection ID:	CO001682
Collection Summary:	Cells were grown at 27°C, 210 rpm in 5 mL of complete synthetic medium (CSM, 0.67% Nitrogen base without amino acids and ammonium sulfate, 2% glucose). The next day, cells were replaced with fresh CSM and grow at 27°C to reach an OD of 0.6 - 0.7. 25 ml of each sample were harvested by centrifugation at 3,838 x g for 5 min at 4°C, washed with 1 ml ice cold sterile water, quenched using ice cold methanol, and stored at -80°C.
Sample Type:	Yeast cells

Treatment:

Treatment ID:	TR001702
Treatment Summary:	No treatment

Sample Preparation:

Sampleprep ID:	SP001695
Sampleprep Summary:	Extraction of metabolites was performed by homogenization in 1 mL of cold methanol/H2O (1:1) extraction solution and centrifugated at 167 x g at 4°C for 5 min. Supernatants were collected and evaporated to dryness in a nitrogen stream stream at 50 ºC (RapidVap, Labconco), and derivatized by methoxyamination by adding 50 μl of 20 mg/ml solution of methoxyamine hydrochloride in pyridine followed by incubation at 37°C for 2 hours. Trimethylsilylation was subsequently performed by adding 50 µl of N-methyl-N-trimethylsilyl-trifluoroacetamide (MSTFA+1% TMCS) and incubated for 1 hour at 65°C. Samples were centrifuged at 13000 rpm for 10 minutes at RT. Supernatants were transferred to glass vials for GC/MS analysis

Sampleprep ID:

SP001695

Sampleprep Summary:

Extraction of metabolites was performed by homogenization in 1 mL of cold methanol/H2O (1:1) extraction solution and centrifugated at 167 x g at 4°C for 5 min. Supernatants were collected and evaporated to dryness in a nitrogen stream stream at 50 ºC (RapidVap, Labconco), and derivatized by methoxyamination by adding 50 μl of 20 mg/ml solution of methoxyamine hydrochloride in pyridine followed by incubation at 37°C for 2 hours. Trimethylsilylation was subsequently performed by adding 50 µl of N-methyl-N-trimethylsilyl-trifluoroacetamide (MSTFA+1% TMCS) and incubated for 1 hour at 65°C. Samples were centrifuged at 13000 rpm for 10 minutes at RT. Supernatants were transferred to glass vials for GC/MS analysis

Combined analysis:

Analysis ID	AN002647
Analysis type	MS
Chromatography type	GC
Chromatography system	Shimadzu GCMS-QP2010 ultra
Column	Shimadzu SH-Rxi-5ms (30m x 0.25mm,0.25um)
MS Type	EI
MS instrument type	Single quadrupole
MS instrument name	Shimadzu QP2010 Ultra
Ion Mode	POSITIVE
Units	mM

Chromatography:

Chromatography ID:	CH001955
Instrument Name:	Shimadzu GCMS-QP2010 ultra
Column Name:	Shimadzu SH-Rxi-5ms (30m x 0.25mm,0.25um)
Internal Standard:	1 mM 2-Fluobiphenyl
Chromatography Type:	GC

MS:

MS ID:	MS002459
Analysis ID:	AN002647
Instrument Name:	Shimadzu QP2010 Ultra
Instrument Type:	Single quadrupole
MS Type:	EI
MS Comments:	Inlet temperature was 280 °C; ion source temperature was 200 °C. MS conditions were set as follows: full scan mode, electron energy of 70 eV, quadrupole scan range of m/z 35–700. Data were processed using the GCMS Solution Postrun Analysis software (Shimadzu Inc) for metabolites identification from their electron impact mass spectra by comparison to the NIST 2014 spectral mass library.
Ion Mode:	POSITIVE