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MB Sample ID: SA257864

Local Sample ID:MCF000887_SR06
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

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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 IDMB Sample IDFactor Level IDLevel ValueFactor Name
MCF000887_SR06SA257864FL033100PMM2-CDGGenotype
MCF000887_SR06SA257864FL0331005.5mM 13C Glc siRNATreatment

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
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