Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA078060

Local Sample ID:	BTWT Cecal_3
Subject ID:	SU001188
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6 germ-free mice
Animal Animal Supplier:	Taconic

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Subject:

Subject ID:	SU001188
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6 germ-free mice
Animal Animal Supplier:	Taconic

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
BTWT Cecal_3	SA078060	FL011887	Bacteroides thetaiotaomicron	Inoculum
BTWT Cecal_3	SA078060	FL011887	WT (VPI-5482)	Strain

Collection:

Collection ID:	CO001182
Collection Summary:	Caecal samples from GF mice colonized with B. thetaiotaomicron WT or the ΔSPT mutant were weighed, and 20 mg of each wet-weight sample was used for lipid profiling.
Sample Type:	Cecum
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR001203
Treatment Summary:	In an anaerobic chamber, bacterial cultures from frozen stocks were first plated on BHI supplemented with hemin and vitamin K, and subsequently pure cultures were selected and mixed together at a 1:1 ratio in sterile, reduced PBS. Bacterial mixtures in PBS were removed from the anaerobic chamber and immediately transported to the animal facility for gavage experiments. The volume of the mixture received per mouse was 100 μL, at a concentration of 10^9 cells/mL. The concentration of the mixture in cell/mL was determined using a UV spectrometer, and gavage doses were confirmed by back-titering the inocula.

Treatment ID:

TR001203

Treatment Summary:

In an anaerobic chamber, bacterial cultures from frozen stocks were first plated on BHI supplemented with hemin and vitamin K, and subsequently pure cultures were selected and mixed together at a 1:1 ratio in sterile, reduced PBS. Bacterial mixtures in PBS were removed from the anaerobic chamber and immediately transported to the animal facility for gavage experiments. The volume of the mixture received per mouse was 100 μL, at a concentration of 10^9 cells/mL. The concentration of the mixture in cell/mL was determined using a UV spectrometer, and gavage doses were confirmed by back-titering the inocula.

Sample Preparation:

Sampleprep ID:	SP001196
Sampleprep Summary:	20 mg of each wet-weight cecal sample was extracted using 200 μL isopropanol + 0.1 ng/μL C24:0 PC as internal standard, incubated at RT for 1 hr, and centrifuged for 10 min at 10,000g.

Combined analysis:

Analysis ID	AN001852
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Shimadzu Nexera X2
Column	Waters Acquity BEH C8 (100 x 2.1mm,1.7um)
MS Type	ESI
MS instrument type	Orbitrap
MS instrument name	Thermo Q Exactive Focus
Ion Mode	POSITIVE
Units	abundance

Chromatography:

Chromatography ID:	CH001340
Instrument Name:	Shimadzu Nexera X2
Column Name:	Waters Acquity BEH C8 (100 x 2.1mm,1.7um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS001713
Analysis ID:	AN001852
Instrument Name:	Thermo Q Exactive Focus
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	Raw data were processed using TraceFinder 3.1 (Thermo Fisher) and Progenesis QI (Nonlinear dynamics). Retention time, m/z, MS/MS fragmentation were used to confirm the identity of metabolite using authentic reference standards when available (level 1 identifications), in addition to monitoring the product ions produced and the retention time and mass of species belonging to the each lipid class (level 2-3 annotation).
Ion Mode:	POSITIVE