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MB Sample ID: SA137925
| Local Sample ID: | Fru1 |
| Subject ID: | SU001699 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001699 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Fru1 | SA137925 | FL017545 | Fructose | Treatment |
Collection:
| Collection ID: | CO001692 |
| Collection Summary: | Isolation and Culture of Mouse Intestinal Organoids |
| Sample Type: | Intestine |
Treatment:
| Treatment ID: | TR001712 |
| Treatment Summary: | For metabolic flux analysis of 13C-labeled glucose, intestinal organoids were pretreated with unlabeled fructose (10 mM) for 24 hr, washed with PBS, and then switched into 13C labeled glucose-containing DMEM/F12 medium (10 mM U-[13C] D-glucose, Cambridge Isotope Laboratories) for 6 hr. To trace the metabolism of fructose, intestinal organoids were cultured in unlabeled fructose-containing DMEM/F12 medium (10 mM D-fructose, 0 mM D-glucose, 2.5 mM L-glutamine) for 12 hr, and then switched into 13C labeled fructose-containing medium (10 mM U-[13C] D-fructose, Sigma-aldrich) for 6 hr. |
Sample Preparation:
| Sampleprep ID: | SP001705 |
| Sampleprep Summary: | To harvest intracellular metabolites, organoid samples were washed with pre-cooled PBS for three times and metabolism was quenched with 80% ice-cold methanol. Pre-cooled methanol (1mL) with para-chlorophenylalanine (1 µM) as the internal standard was added to each sample and incubated on ice for 10 min. Sample lysates were then centrifuged at 4 °C for 15 min at 23,000 g. |
Combined analysis:
| Analysis ID | AN002657 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | UPLC-Q-TOF-MS (Synapt G2si, waters) |
| Column | Waters XBridge Amide (100 x 4.6mm,3.5um) |
| MS Type | ESI |
| MS instrument type | GC-TOF |
| MS instrument name | Waters Synapt G2 Si QTOF |
| Ion Mode | NEGATIVE |
| Units | abundance |
Chromatography:
| Chromatography ID: | CH001960 |
| Instrument Name: | UPLC-Q-TOF-MS (Synapt G2si, waters) |
| Column Name: | Waters XBridge Amide (100 x 4.6mm,3.5um) |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS002464 |
| Analysis ID: | AN002657 |
| Instrument Name: | Waters Synapt G2 Si QTOF |
| Instrument Type: | GC-TOF |
| MS Type: | ESI |
| MS Comments: | MS MassLynx v4.1 Quanlynx v4.1 |
| Ion Mode: | NEGATIVE |
