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MB Sample ID: SA016170

Local Sample ID:Blood444
Subject ID:SU000384
Subject Type:mouse
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57Bl/6
Age Or Age Range:4.5 mo
Weight Or Weight Range:29.1 g-49.2 g
Gender:male
Animal Animal Supplier:JAX
Animal Housing:4/cage
Animal Feed:Purina chow
Species Group:Mammal

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Sample Preparation:

Sampleprep ID:SP000391
Sampleprep Summary:For extraction, 500 µL of an ice-cold solution of 90:10 Methanol:Chloroform (v/v) ¬was added to the tubes containing the thawed cell pellets. MagNA lyser ceramic beads were added to the tubes, and a MagNA lyser was used to beat the samples for two 30 seconds pulses at 2,000 rpm placing samples on cold block for 5 min in between pulses. Sonication for 5 min was performed to the beaten cell samples followed by centrifugation at room temperature and at 16,000rcf for 4 min. A 350 µL aliquot of each experimental cell sample homogenized supernatant was transferred to a labeled 2.0 mL Lo-Bind Eppendorf tube. Analytical pooled samples were created by combining a 125 µL aliquot from each experimental sample into a labeled 20 mL glass vial. The QC pooled sample was vortexed for 30 sec and 350 µL aliquots were transferred to 5 labeled 2.0 mL Lo-Bind eppendorf tubes. Next, the supernatants of the study and QC pooled samples were dried overnight using a lyophilizer. The residue was reconstituted in 50 µL of reconstitution buffer (95:5 Water:Methanol, v/v) and mixed on an analog vortex mixer for 1 min with speed set at 10. Then, the samples were centrifuged at room temperature for 4 min at 16,000 rcf, and the supernatants were transferred to autosampler vials for data acquisition.
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