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MB Sample ID: SA257906
| Local Sample ID: | MCF0000858_SR09 |
| Subject ID: | SU002664 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Genotype Strain: | WT/PMM2-CDG |
| Age Or Age Range: | 5-45 |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002664 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Genotype Strain: | WT/PMM2-CDG |
| Age Or Age Range: | 5-45 |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| MCF0000858_SR09 | SA257906 | FL033108 | WT | Genotype |
| MCF0000858_SR09 | SA257906 | FL033108 | 5.5mM 13C Glc siRNA | Treatment |
Collection:
| Collection ID: | CO002657 |
| Collection Summary: | Cells were washed with PBS, cells were incubated with extraction buffer for 2min before scraping and transferring to a fresh eppendorf. Samples were precipitated overnight at -80, then they were centrifuged at max rpm, 20min, 4 degrees C and supernatant transferred to an M/S vial. |
| Sample Type: | Fibroblasts |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002676 |
| Treatment Summary: | Cells were treated with vehicle or 5nM siRNA targeting AKR1B1 |
Sample Preparation:
| Sampleprep ID: | SP002670 |
| Sampleprep Summary: | Metabolites were extracted with 80% Methanol and IS (d27 myristic acid) |
| Processing Storage Conditions: | -80℃ |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN004224 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters Acquity |
| Column | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | NEGATIVE |
| Units | AUC |
Chromatography:
| Chromatography ID: | CH003133 |
| Chromatography Summary: | C18 iP REVERSE PHASE |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| Column Temperature: | 40 |
| Flow Gradient: | The gradient started with 5% of solvent B and 95% solvent A and remained at 5% B until 2 min post injection. A linear gradient to 37% B was carried out until 7 min and increased to 41% until 14 min. Between 14 and 26 minutes the gradient increased to 95% of B and remained at 95% B for 4 minutes. At 30 min the gradient returned to 5% B. The chromatography was stopped at 40 min. |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 100% water; 10mM tributylamine; 15mM acetic acid |
| Solvent B: | 100% methanol |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003971 |
| Analysis ID: | AN004224 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | El-Maven polly, ThermoFisher Xcalibur; Metabolites were annotated using the inhouse standard metabolite library- elution time and m/z values. |
| Ion Mode: | NEGATIVE |
