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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 |
