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MB Sample ID: SA194292
Local Sample ID: | KP_4 |
Subject ID: | SU002148 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57Bl/6 GEMMs |
Age Or Age Range: | Mutants: K: 90 days, KK: 60 days, KL and KKL: 40 days post Ad5-CMV-Cre |
Gender: | Male and female |
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Sample Preparation:
Sampleprep ID: | SP002154 |
Sampleprep Summary: | Tumor tissue was flash frozen and crushed using a mortar and pestle in liquid nitrogen. To the Eppendorf tubes 150 µL of cold 80 % acetonitrile was added and vortexed quickly. The suspension was transferred to a fresh Eppendorf tube. Another 150 µL portion of extraction solvent was added to the tubes, vortexed and the suspension combined with the first portion. To the combined extraction mixture 75 µL of CHCl3 was added and samples were mixed for 1 h in a cool room to ensure complete extraction. The samples were centrifuged at 4 °C at 14.8 g for 10 min, supernatant transferred to new tubes and evaporated at 20 °C under a stream of nitrogen. BSA assay was performed on the protein pellet by dissolving all protein in equal volume of detergent solution (4 % v/v SDS in water). Dried samples were resolubilized in appropriate volume of CHCl3 : MeOH : H2O (1 : 3 :1, v/v) mixture based on measured protein content, shaken for 15 min at room temperature, centrifuged at 4 °C at 14.8 g for 10 min. 100 µL of the supernatant was transferred to LCMS vials. 10 µL injected for metabolomics analysis. |
Processing Storage Conditions: | -80℃ |
Extract Storage: | -80℃ |