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MB Sample ID: SA185839
Local Sample ID: | RH2-12 |
Subject ID: | SU002067 |
Subject Type: | Cultured cells |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | Fbxo7LacZ mice (Fbxo7tm1a(EUCOMM)Hmgu on a C57BL/6J background) were bred as heterozygous crosses |
Age Or Age Range: | WT, heterozygous and homozygous littermates were harvested between 6-9 weeks. |
Gender: | Male and female |
Subject Comments: | All experiments in mice were performed in accordance with the UK Animals (Scientific Procedures) Act 1986 and ARRIVE guidelines. Animal licences were approved by the Home Office and the University of Cambridge's Animal Welfare & Ethical Review Body Standing Committee. Experiments were performed under Home Office licence PPL 70/9001. |
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Sample Preparation:
Sampleprep ID: | SP002073 |
Sampleprep Summary: | To extract metabolites, cells were harvested, washed twice in PBS and resuspended in 200 μL ice cold metabolite extraction solution (50% LC–MS grade methanol, 30% LC-MS grade acetonitrile, 20% ultrapure water, 5 μM valine-d8 as internal standard) per 1x106 cells. Cells were incubated in a dry ice-methanol bath for 20 min, then at 4oC with shaking for 15 min. Samples were centrifuged at 13000 rpm for 20 min and the supernatant was collected into autosampler vials for LC-MS analysis. |