Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA136744

Local Sample ID:	YGR023w-4
Subject ID:	SU001686
Subject Type:	Yeast
Subject Species:	Saccharomyces cerevisiae
Taxonomy ID:	4932
Genotype Strain:	BY4742, YGR023w
Cell Biosource Or Supplier:	Open Biosystems

Select appropriate tab below to view additional metadata details:

Subject:

Subject ID:	SU001686
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	SA136744	FL017498	mtl1Δ	Strain

Collection:

Collection ID:	CO001679
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:	TR001699
Treatment Summary:	No treatment

Sample Preparation:

Sampleprep ID:	SP001692
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). Nitrilation was performed by adding 150 μL of 0.2 mM hydroxylammonium chloride in pyridine to the dried sample, and then heating at 90ºC for 40 minutes. After that, acetylation was performed by adding 250 μL of acetic anhydride, and then heating at 90 ºC for 60 minutes. After that, the sample was dried again in a nitrogen gas stream, and then redissolved in 400 μL of ethyl acetate. Derivatized samples were collected and stored at -20ºC.

Sampleprep ID:

SP001692

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). Nitrilation was performed by adding 150 μL of 0.2 mM hydroxylammonium chloride in pyridine to the dried sample, and then heating at 90ºC for 40 minutes. After that, acetylation was performed by adding 250 μL of acetic anhydride, and then heating at 90 ºC for 60 minutes. After that, the sample was dried again in a nitrogen gas stream, and then redissolved in 400 μL of ethyl acetate. Derivatized samples were collected and stored at -20ºC.

Combined analysis:

Analysis ID	AN002643
Analysis type	MS
Chromatography type	GC
Chromatography system	Shimadzu GCMS-QP2010 ultra
Column	Shimadzu SH-RXI (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:	CH001952
Instrument Name:	Shimadzu GCMS-QP2010 ultra
Column Name:	Shimadzu SH-RXI (30m x 0.25mm,0.25um)
Chromatography Type:	GC

MS:

MS ID:	MS002455
Analysis ID:	AN002643
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: SIM acquisition mode, electron energy of 70 eV, quadrupole scan range of m/z 115; 187; 212;314. 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