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MB Sample ID: SA137117
Local Sample ID: | 9--3 |
Subject ID: | SU001691 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | BALB/c |
Age Or Age Range: | 70-74 |
Gender: | Male and female |
Animal Animal Supplier: | Montana State University Animal Resource Center |
Animal Feed: | PicoLab Rodent Diet 20 5053 |
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Sample Preparation:
Sampleprep ID: | SP001697 |
Sampleprep Summary: | Frozen livers were homogenized in liquid nitrogen using a mortar and pestle. Liver powder was weighed (60-80 mg) into 2-mL microcentrifuge tubes. 900 µL of ice-cold 1:2 MeOH/CHCl3 was added to the liver tissue and tissue was mixed into suspension and lysed using a FastPrep-24TM 5G homogenizer using the S. aureus setting of 6.0 meters/second speed for 2 x 40 seconds. Tubes were placed back on ice and 200 µL of ice-cold H2O was added, before repeating the lysis step. Samples were then placed at -20 ºC for approximately 45 minutes to precipitate protein, then centrifuged at 14,000 g for 10 minutes. 250 µL of upper polar phase were transferred to a 1.5 mL Eppendorf tube and dried overnight using a Savant™ Universal SpeedVac™ vacuum system with no heat. Dried metabolite mixtures were re-suspended in 600 µL of NMR buffer containing 0.25 mM 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS), 90%H2O/10% D2O, 25 mM sodium phosphate, pH 7. |