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MB Sample ID: SA314481
| Local Sample ID: | 13 |
| Subject ID: | SU002989 |
| Subject Type: | Mammal |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002989 |
| Subject Type: | Mammal |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 13 | SA314481 | FL037448 | Low NSD2 | Genotype |
| 13 | SA314481 | FL037448 | Knockdown 1 | Treatment |
Collection:
| Collection ID: | CO002982 |
| Collection Summary: | Multiple myeloma cells were collected, spun down, and flash frozen. |
| Sample Type: | B cells |
Treatment:
| Treatment ID: | TR002998 |
| Treatment Summary: | Stable NSD2 knockdown cell lines and control cell lines were established using shRNA. |
Sample Preparation:
| Sampleprep ID: | SP002995 |
| Sampleprep Summary: | Soluble metabolites were extracted directly from snap-frozen cell pellets using cold acetonitrile/water (80/20, vol/vol) at a dilution of 10,000 cells/ul solvent. Samples were vortexed and incubated at -80C overnight to precipitate proteins and subsequently thawed, vortexed, and centrifuged at 18,000 x g for 30 min at 4C to pellet debris. The supernatants were transferred to new vials and analyzed by high-performance liquid chromatography (HPLC) and high-resolution mass spectrometry and tandem mass spectrometry (MS/MS). |
Combined analysis:
| Analysis ID | AN004713 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 |
| Column | Waters XBridge BEH Amide (100 x 4.6mm,3.5um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH003549 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters XBridge BEH Amide (100 x 4.6mm,3.5um) |
| Column Temperature: | 275 |
| Flow Gradient: | 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1-18 min, 75% A; 18-25 min, 15% A |
| Flow Rate: | 150 μL/min |
| Solvent A: | 95% water/5% acetonitrile; 10 mM ammonium hydroxide; 10 mM ammonium acetate, pH 9.0 |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004459 |
| Analysis ID: | AN004713 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The capillary of the ESI source was set to 275 °C, with sheath gas at 35 arbitrary units, auxiliary gas at 5 arbitrary units and the spray voltage at 4.0 kV. In positive/negative polarity switching mode, an m/z scan range from 60 to 900 was chosen and MS1 data was collected at a resolution of 70,000. |
| Ion Mode: | UNSPECIFIED |
