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MB Sample ID: SA358127
Local Sample ID: | control3 |
Subject ID: | SU003421 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Gender: | Male |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU003421 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Gender: | Male |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
control3 | SA358127 | FL041791 | Blood serum | Sample source |
control3 | SA358127 | FL041791 | WT | Genotype |
control3 | SA358127 | FL041791 | control | Treatment |
Collection:
Collection ID: | CO003414 |
Collection Summary: | Blood was collected from control or overexpression of SLC7A14 in POMC neuron mice.Blood sample was collected into a BD Vacutainer tube not containing any anticoagulant, which was allowed to sit for ~30–60 min for clots to form following which serum was obtained by centrifugation (15 min at 4°C at 3500 rpm). |
Sample Type: | Blood (serum) |
Treatment:
Treatment ID: | TR003430 |
Treatment Summary: | To overexpression of SLC7A14 in ARC POMC neurons, POMC Cre mice were bilaterally injected either with a Cre-dependent AAV vector containing SLC7A14 in the opposite orientation flanked by two inverted loxP sites (AAV9-Syn-DIO-SLC7A14-mCherry, 1.5 × 1012 Pfu/mL, HANBIO) at a volume of 200 nL into the ARC or an AAV vector containing only mCherry in the opposite orientation flanked by two inverted loxP sites (AAV9-Syn-DIO-mCherry, 1.5 × 1012 Pfu/mL, HANBIO) as a control. Serum was collected from control or overexpression of SLC7A14 in POMC neuron mice. |
Sample Preparation:
Sampleprep ID: | SP003428 |
Sampleprep Summary: | The samples (100 μL) were placed in the EP tubes and resuspended with prechilled 80% methanol and 0.1% formic acid by well vortex. Then the sampleswere incubated on ice for 5 min and centrifuged at 15,000 g, 4°C for 20 min. Some of supernatant was diluted to final concentration containing 53% methanol by LC-MS grade water.The samples were subsequently transferred to a fresh Eppendorf tube and then were centrifuged at 15000 g, 4°C for 20 min. Finally, the supernatant was injected into the LC-MS/MS system analysis |
Combined analysis:
Analysis ID | AN005407 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Thermo Vanquish |
Column | Thermo Hypersil GOLD aQ (100 x 2.1mm,1.9um) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Q Exactive HF-X Orbitrap |
Ion Mode | NEGATIVE |
Units | Peak area |
Chromatography:
Chromatography ID: | CH004100 |
Instrument Name: | Thermo Vanquish |
Column Name: | Thermo Hypersil GOLD aQ (100 x 2.1mm,1.9um) |
Column Temperature: | 40°C |
Flow Gradient: | 2% B, 1.5 min; 2-100% B, 12.0 min; 100% B, 14.0 min;100-2% B, 14.1 min;2% B, 17 min. |
Flow Rate: | 0.2 mL/min |
Solvent A: | 100% Water; 5mM Ammonium acetate |
Solvent B: | 100% Methanol |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS005134 |
Analysis ID: | AN005407 |
Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | The raw data files generated by UHPLC-MS/MS were processed using the Compound Discoverer 3.1 (CD3.1, ThermoFisher) to perform peak alignment, peak picking, and quantitation for each metabolite. The main parameterswere set as follows: retention time tolerance, 0.2 minutes; actual mass tolerance, 5ppm; signal intensity tolerance, 30%; signal/noise ratio, 3; and minimum intensity, et al. After that, peak intensities were normalized to the total spectral intensity.The normalized data was used to predict the molecular formula based on additive ions, molecular ion peaks and fragment ions. And then peaks were matched with the mzCloud (https://www.mzcloud.org/),mzVault and MassList database to obtain the accurate qualitative and relative quantitative results.Statistical analyses were performed using the statistical software R (R version R-3.4.3),Python (Python 2.7.6 version) and CentOS (CentOS release 6.6),When data were not normally distributed, normal transformations were attempted using of area normalization method. |
Ion Mode: | NEGATIVE |