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MB Sample ID: SA073436
Local Sample ID: | 786-O rep3 |
Subject ID: | SU001130 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001130 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
786-O rep3 | SA073436 | FL011401 | Wild-type | Genotype |
Collection:
Collection ID: | CO001124 |
Collection Summary: | Quickly aspirate cell culture medium from 3.5 cm plates, gently dispense 2 mL of PBS to rinse cells, shake the plate back and forth for 2 s and then quickly remove the PBS. Immediately add 1 mL 8:2 methanol:H2O (on dry ice) into the plates placed on dry ice to quench cell metabolism, store samples at -80℃. |
Sample Type: | Cultured cells |
Treatment:
Treatment ID: | TR001144 |
Treatment Summary: | The cell lines including 786-O and 38E/38F were cultured in RPMI 1640 medium supplemented with 10% FBS, 1% penicillin/streptomycin, and 2 mM L-glutamine. After culture at 37 °C, 5% CO2 condition for 24 h, the cells were collected for GC-MS analysis. |
Sample Preparation:
Sampleprep ID: | SP001137 |
Sampleprep Summary: | Sampleprep Summary: Scrape all the cells from culture dishes on dry ice, and transfer them into a 2 mL Eppendorf, spin the mixture at 14000 rpm for 10 min at 4°C. Remove all the soluble extract into a vial and completely dry samples. The extracted samples were incubated with 30 μL O-methylhydroxylamine hydrochloride solution in pyridine (Sigma-Aldrich, St. Louis, MO, USA) at 60°C for 90 min. Next, 70 μL of N-tert-Butyldimethylsilyl-N-methyltrifluoroacetamide (MTBSTFA) were added and placed at 60 °C for 30 min. After centrifuge at 14000 rpm for 10 min, 70 μL of supernatant was transferred to a clean glass GC insert tube. |
Combined analysis:
Analysis ID | AN001771 |
---|---|
Analysis type | MS |
Chromatography type | GC |
Chromatography system | Agilent 7890A |
Column | Agilent DB5-MS (30m × 0.25mm, 0.25um) |
MS Type | EI |
MS instrument type | Single quadrupole |
MS instrument name | Agilent 5975C |
Ion Mode | POSITIVE |
Units | pmole/ug |
Chromatography:
Chromatography ID: | CH001251 |
Instrument Name: | Agilent 7890A |
Column Name: | Agilent DB5-MS (30m × 0.25mm, 0.25um) |
Flow Rate: | 0.5 mL/min |
Injection Temperature: | 250°C |
Transferline Temperature: | 290°C |
Chromatography Type: | GC |
MS:
MS ID: | MS001637 |
Analysis ID: | AN001771 |
Instrument Name: | Agilent 5975C |
Instrument Type: | Single quadrupole |
MS Type: | EI |
Ion Mode: | POSITIVE |
Ion Source Temperature: | 230°C |
Ionization: | 70 eV |
Scanning Range: | 50-600 m/z |