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MB Sample ID: SA211364
Local Sample ID: | S9 |
Subject ID: | SU002290 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | wild type and endothelium-specific Sirt1 knockout mice |
Age Or Age Range: | 12 weeks old |
Weight Or Weight Range: | ~30 g |
Height Or Height Range: | n/a |
Gender: | Male |
Animal Animal Supplier: | Jax |
Animal Housing: | University of Iowa animal house |
Animal Light Cycle: | 12 h light/12 h night |
Animal Feed: | Standard Chow |
Animal Water: | Tap water |
Animal Inclusion Criteria: | n/a |
Species Group: | n/a |
Select appropriate tab below to view additional metadata details:
Sample Preparation:
Sampleprep ID: | SP002296 |
Sampleprep Summary: | Tissue samples were lyophilized overnight prior to bead mill homogenization in 18:1 (µl:mg wet tissue weight) ice-cold 2:2:1 methanol:acetonitrile:water extraction buffer containing a mixture of internal standards (D4-citric acid, D4-succinic acid, D8-valine, and U13C-labeled glutamine, glutamic acid, lysine, methionine, serine, and tryptophan; Cambridge Isotope Laboratories). Homogenates were rotated for 1 hour at -20°C. Homogenates were centrifuged for 10 minutes at 21,000 x g, and 150 µl of the cleared metabolite extracts were transferred to autosampler vials and dried using a SpeedVac vacuum concentrator (Thermo). Dried metabolite extracts were reconstituted in 30 μl of 11.4 mg/ml methoxyamine (MOX) in anhydrous pyridine, vortexed for 5 minutes, and heated for 1 hour at 60°C. Next, to each sample 20 μl of N,O-Bis(trimethylsilyl)trifluoroacetamide (TMS) was added, samples were vortexed for 1 minute, and heated for 30 minutes at 60°C. |
Sampleprep Protocol Filename: | GC_Method_Tissue.pdf |