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MB Sample ID: SA204323

Local Sample ID:BM_supernatant_Ctrl_1
Subject ID:SU002210
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

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Subject:

Subject ID:SU002210
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
BM_supernatant_Ctrl_1SA204323FL025077Bone marrow supernatantSource_Name[gating]
BM_supernatant_Ctrl_1SA204323FL025077HSC BasalTreatment
BM_supernatant_Ctrl_1SA204323FL0250771cBatch
BM_supernatant_Ctrl_1SA204323FL025077Unlabeled AAs were used for total AA (FigS2.D)Note

Collection:

Collection ID:CO002203
Collection Summary:5×10^5 indicated cells were incubated with 100 µM stable [13C,15N] amino acids (MSK-A2-US-1.2, Cambridge Isotope Laboratories) for indicated time and centrifuged at 500 g for 5 min at 4 °C. The pelleted cells were extracted in 500 µl ice-cold Acetonitrile: Isopropyl Alcohol: water (3:3:2 v/v/v) and aliquoted as three technical replicates. The extracts were vortexed for 5 min at 4 °C and centrifuged at 14,000 g for 2 min at 4 °C. The supernatants were dried by vacuum spin for subsequent derivatization and stored at -20 ℃.
Sample Type:Bone marrow

Treatment:

Treatment ID:TR002222
Treatment Summary:Germ-free C57BL/6J mice were intraperitoneally injected with 10 mg/kg 5FU (F6627-5G, Sigma-Aldrich) for 14 days, or fed with vehicle or NR (400 mg/kg per day) (1341-23-7, ziyi-reagent) for consecutive 8 weeks (long-term) or 1 week (short-term). Hematopoietic cells were stained and sorted from treated mice.

Sample Preparation:

Sampleprep ID:SP002216
Sampleprep Summary:To get the corresponding derivatives, the dried aliquots were incubated with 20 µL 2% (w/v) methoxyamine hydrochloride (226904, Sigma-Aldrich) in pyridine for 60 min at 37 °C, and silylated by 30 µL of N-Methyl-N-(tert-butyldimethylsilyl) trifluoroacetamide with 1% tert-Butyldimethylchlorosilane (TBDMS, 18162-48-6, Regis Technologies) for 30 min at 45 °C. The corresponding derivatives were analyzed by GC-MS using the Trace 1310 gas chromatograph (Thermo Fisher) with the DB-35ms column (Agilent Technologies) connected to the Q ExactiveTM GC OrbitrapTM GC-MS/MS system (Thermo Fisher).

Combined analysis:

Analysis ID AN003478
Analysis type MS
Chromatography type GC
Chromatography system TRACE 1310
Column DB-35ms
MS Type EI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Orbitrap
Ion Mode POSITIVE
Units Peak Area

Chromatography:

Chromatography ID:CH002567
Instrument Name:TRACE 1310
Column Name:DB-35ms
Chromatography Type:GC

MS:

MS ID:MS003239
Analysis ID:AN003478
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:EI
MS Comments:GC-MS data was acquired on Q-Exactive Orbitrap mass spectrometer (Thermo Fisher) coupled with Gas Chromatograph system TRACE 1310 (Thermo Fisher). Chromatographic separation was performed on a DB-35ms column module (30 m length x 0.25 mm internal diameter, Agilent Technologies). The column temperature was programmed with an initial temperature of 50 °C for 2 min, then ramped at 10°C/min to 325 °C, and maintained for 5 min. The mass range was set as 50-600 m/z, and the resolution was 60,000. The ion source temperature was 300°C with the transfer line temperature of 250°C, and the electron energy was 70 eV with EI source.
Ion Mode:POSITIVE
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