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MB Sample ID: SA136771
Local Sample ID: | Sample2-NoISD |
Subject ID: | SU001688 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001688 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Sample2-NoISD | SA136771 | FL017502 | Skeletal muscle | Treatment |
Collection:
Collection ID: | CO001681 |
Collection Summary: | 5 skeletal muscle and 5 sarcoma samples. Tissue polar metabolites were extracted into 80% methanol/water. |
Sample Type: | Blood;Sarcoma;Skeletal muscle |
Treatment:
Treatment ID: | TR001701 |
Treatment Summary: | The mouse model of soft-tissue sarcoma was generated on a mixed background (129/SvJae and C57BL/6) using a combination of alleles that were previously described: Pax7CreER-T2 20, p53FL/FL 21, LSL-NrasG12D 22 and ROSA26mTmG Primary mouse soft tissue sarcomas were generated in the mouse hind limb as previously described 19 by intramuscular (IM) injection of (Z)-4-hydroxytamoxifen (4-OHT). 4-OHT was dissolved in 100% DMSO at a concentration of 10 mg/ml and 50 ul of the solution was injected into the gastrocnemius muscle. |
Sample Preparation:
Sampleprep ID: | SP001694 |
Sampleprep Summary: | The tumor sample was first homogenized in liquid nitrogen and then 5 to 10 mg was weighed in a new Eppendorf tube. Ice cold extraction solvent (250 ul 80% MeOH/water) was added to the tissue sample, and a pellet mixer was used to further break down the tissue chunk and form an even suspension, followed by the addition of 250 ul to rinse the pellet mixer. After incubation on ice for an additional 10 min, the tissue extract was centrifuged with a speed of 20,000 g at 4 °C for 10 min. The dry pellets were reconstituted into 30 ul (per 3 mg tissue) sample solvent (water:methanol: acetonitrile, 2:1:1, v/v), and 3 ul was injected to LC-HRMS. 5 ul mouse plasma was mixed with 5 ul water, and 40 ul ice cold methanol was added. After vortex for 1 min, the mixture was centrifuged with a speed of 20,000 g at 4 °C for 10 min, and 3 l was injected to LC-HRMS. |
Combined analysis:
Analysis ID | AN002645 | AN002646 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | HILIC | HILIC |
Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
Column | Waters Xbridge amide (100 x 2.1mm,3.5um) | Waters Xbridge amide (100 x 2.1mm,3.5um) |
MS Type | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap |
MS instrument name | Thermo Exactive Plus Orbitrap | Thermo Exactive Plus Orbitrap |
Ion Mode | POSITIVE | NEGATIVE |
Units | peak area | peak area |
Chromatography:
Chromatography ID: | CH001954 |
Chromatography Summary: | HILIC method is for general metabolomics analysis. |
Instrument Name: | Thermo Dionex Ultimate 3000 |
Column Name: | Waters Xbridge amide (100 x 2.1mm,3.5um) |
Chromatography Type: | HILIC |
MS:
MS ID: | MS002457 |
Analysis ID: | AN002645 |
Instrument Name: | Thermo Exactive Plus Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | It was operated in the full-scan mode with the resolution set at 70 000 (at m/z 200).LC-MS data were analyzed using Sieve 2.0 (Thermo Scientific), and the integrated area under metabolite peak was used to compare the relative abundance of each metabolite in different samples in the same batch. |
Ion Mode: | POSITIVE |
MS ID: | MS002458 |
Analysis ID: | AN002646 |
Instrument Name: | Thermo Exactive Plus Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | It was operated in the full-scan mode with the resolution set at 70 000 (at m/z 200).LC-MS data were analyzed using Sieve 2.0 (Thermo Scientific), and the integrated area under metabolite peak was used to compare the relative abundance of each metabolite in different samples in the same batch. |
Ion Mode: | NEGATIVE |