Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA069160

Local Sample ID:	V1E2T1P
Subject ID:	SU001071
Subject Type:	Other
Subject Species:	Xenopus laevis
Taxonomy ID:	8355
Age Or Age Range:	Embryos were obtained from natural mating of frogs (Nasco)
Weight Or Weight Range:	Sexually mature male and female frogs
Gender:	Not applicable

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Subject:

Subject ID:	SU001071
Subject Type:	Other
Subject Species:	Xenopus laevis
Taxonomy ID:	8355
Age Or Age Range:	Embryos were obtained from natural mating of frogs (Nasco)
Weight Or Weight Range:	Sexually mature male and female frogs
Gender:	Not applicable

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
V1E2T1P	SA069160	FL011032	WT	Embryo Type

Collection:

Collection ID:	CO001065
Collection Summary:	Cells were identified based on morphology, pigmentation, and location in the embryo in comparison to established cell-fate maps for Xenopus laevis embryos. A portion of the identified V1 cell was microaspirated using a fabricated microcapillary.
Collection Protocol ID:	Portero 2018 Metabolomics Workbench Protocols FINAL 2018-08-08
Collection Protocol Filename:	nemes_20180725_143242_PR_SP_Liu_2018_Metabolomics_Workbench_Protocol.pdf
Sample Type:	embryonic cell
Collection Method:	Microaspiration of cell content
Collection Frequency:	1 collection per cell
Collection Duration:	5 s for aspiration
Volumeoramount Collected:	Ca. 10 nL per aspiration
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR001085
Treatment Summary:	All protocols related to the handling and manipulation of animals were approved by the University of Maryland, College Park (College Park, MD). Embryos were dejellied using 2% cystine solution, cultured in 100% Steinberg's solution (media), and used without further treatment.
Treatment Protocol ID:	IACUC # R-DEC-17-57
Treatment Protocol Filename:	nemes_20180725_143242_PR_SP_Liu_2018_Metabolomics_Workbench_Protocol.pdf

Sample Preparation:

Sampleprep ID:	SP001078
Sampleprep Summary:	Ca. 10 nL of cell content were aspirated from identified cells. The aspirated material was ejected into ~4 uL of aqueous mixture of 40% acetonitrile and 40% methanol to extract metabolites.
Sampleprep Protocol Filename:	nemes_20180725_143242_PR_SP_Liu_2018_Metabolomics_Workbench_Protocol.pdf
Processing Method:	On ice, then extracts stored at -80 degC until analysis.
Processing Storage Conditions:	On ice
Extraction Method:	In cold aqueous mixture of 40% acetonitrile and 40% methanol.
Extract Enrichment:	none
Extract Cleanup:	none
Extract Storage:	-80℃
Subcellular Location:	Unknown

Combined analysis:

Chromatography:

Chromatography ID:	CH001191
Chromatography Summary:	Metabolites were separated in a custom-built capillary electrophoresis (CE) system.
Instrument Name:	Custom-built CE system
Column Name:	Bare fused silica capillary
Column Temperature:	Room temperature
Injection Temperature:	Room temperature
Sample Injection:	Ca. 10 nL
Solvent A:	100% water; 1% formic acid
Analytical Time:	45 min of separation
Capillary Voltage:	During cationic separation, +19,000-20,000 V was applied on the inlet end of the CE capillary.
Preconditioning:	Sodium hydroxide solution
Sheath Liquid:	During cationic analysis, the electrospray sheath solution was 50% methanol with 0.1% formic acid.
Chromatography Type:	CE

Chromatography ID:	CH001192
Chromatography Summary:	Metabolites were separated in a custom-built capillary electrophoresis (CE) system.
Instrument Name:	Custom-built CE system
Column Name:	Bare fused silica capillary
Column Temperature:	Room temperature
Injection Temperature:	Room temperature
Sample Injection:	Ca. 10 nL
Solvent A:	100% water; 20 mM ammonium bicarbonate
Analytical Time:	45 min of separation
Capillary Voltage:	During anionic, +19,000-20,000 V was applied on the inlet end of the CE capillary.
Preconditioning:	Sodium hydroxide solution
Sheath Liquid:	During cationic analysis, the electrospray sheath solution was 0.2 mM ammonium bicarbonate in 50% isopropanol.
Chromatography Type:	CE

MS: