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MB Sample ID: SA204263
| Local Sample ID: | KO_2 |
| Subject ID: | SU002208 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002208 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| KO_2 | SA204263 | FL025058 | Mutant | Genotype |
| KO_2 | SA204263 | FL025058 | 5 FU 2 weeks | Treatment |
| KO_2 | SA204263 | FL025058 | 1f | Batch |
| KO_2 | SA204263 | FL025058 | Lin– Scal1+ c-Kit+ (LSK) | Source_Name[gating] |
Collection:
| Collection ID: | CO002201 |
| Collection Summary: | 2 million HSPCs (LSK cells) were sorted stored at -80 ℃. |
| Sample Type: | Bone marrow |
Treatment:
| Treatment ID: | TR002220 |
| Treatment Summary: | Wild-type and GCN2 knockout mice were intraperitoneally injected with 10 mg/kg 5FU (F6627-5G, Sigma-Aldrich) for 14 days. HSPCs were stained and sorted from treated mice. |
Sample Preparation:
| Sampleprep ID: | SP002214 |
| Sampleprep Summary: | The cell samples were extracted with 800 µL of 80% methanol and vortexed for 1 min. Then, ultrasonicate for 30 min at 4℃ were performed, letting stand in -40℃ refrigerator for 1 h. After that, the samples were vortexed for 30s and set at 4℃ for 30 min. The samples were centrifuged at 12,000 rpm, 4℃ for 15 min. Taking all the supernatant in the centrifuge tube and concentrating to remove the organic reagents and water. 100 µL of 80% methanol would be used to reconstitute, vortex again for 1 min. The supernatant was transferred by centrifugation to the sample vial for liquid chromatography-mass spectrometry (LC-MS) analysis. |
Combined analysis:
| Analysis ID | AN003476 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | ACQUITY UPLC |
| Column | Waters ACQUITY UPLC BEH Amide |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | ABI Sciex 5500 QTrap |
| Ion Mode | UNSPECIFIED |
| Units | AU |
Chromatography:
| Chromatography ID: | CH002565 |
| Instrument Name: | ACQUITY UPLC |
| Column Name: | Waters ACQUITY UPLC BEH Amide |
| Column Temperature: | 40 |
| Flow Gradient: | The gradient program started at 85% (B) and was reduced to 70% (B) over 9 min, then reduced from 70% (B) to 30% (B) over 3 min, followed by wash with 30% (B) for another 3 min, and finally 5 min re-equilibration with 85% (B). |
| Flow Rate: | 0.35ml/min |
| Solvent A: | 100% water; 10mM ammonium formate |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003237 |
| Analysis ID: | AN003476 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | LC-MS data was acquired on SCIEX 5500 QQQ mass spectrometer (Applied Biosystems, Foster City, CA, USA) coupled with high-performance liquid chromatography (HPLC) system ACQUITY UPLC (Waters, Milford, MA, USA). Chromatographic separation was performed on a ACQUITY UPLC BEH Amide Column (1.7 µm, 2.1 mm x 100 mm, Waters, Milford, MA, USA). The mobile phase (A) was 10 mM ammonium formate and (B) acetonitrile. The gradient program started at 85% (B) and was reduced to 70% (B) over 9 min, then reduced from 70% (B) to 30% (B) over 3 min, followed by wash with 30% (B) for another 3 min, and finally 5 min re-equilibration with 85% (B). The flow rate was 0.35 mL/min and column compartment temperature was 40ºC. The total run time was 20 min with an injection sample volume of 5 µL. |
| Ion Mode: | UNSPECIFIED |
