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MB Sample ID: SA211362
Local Sample ID: | S7 |
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:
Subject:
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 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
S7 | SA211362 | FL025774 | KO | Genotype |
Collection:
Collection ID: | CO002283 |
Collection Summary: | 40 mg of tibialis anterior was rapidly harvested and frozen immediately in liquid nitrogen. Metabolites were separated, measured, and analyzed. |
Sample Type: | Muscle |
Treatment:
Treatment ID: | TR002302 |
Treatment Summary: | After 5 to 6 h of fasting, mice were anesthetized and skeletal muscle were collected. |
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 |
Combined analysis:
Analysis ID | AN003607 |
---|---|
Analysis type | MS |
Chromatography type | GC |
Chromatography system | Thermo Trace 1310 |
Column | Thermo TraceGold TG-5SilMS |
MS Type | EI |
MS instrument type | Single quadrupole |
MS instrument name | Thermo ISQ |
Ion Mode | POSITIVE |
Units | AU |
Chromatography:
Chromatography ID: | CH002666 |
Chromatography Summary: | 1 μl of derivatized sample was injected into a Trace 1300 GC (Thermo) fitted with a TraceGold TG-5SilMS column (Thermo) operating under the following conditions: split ratio = 20-1, split flow = 24 μl/minute, purge flow = 5 ml/minute, carrier mode = Constant Flow, and carrier flow rate = 1.2 ml/minute. The GC oven temperature gradient was as follows: 80°C for 3 minutes, increasing at a rate of 20°C/minute to 280°C, and holding at a temperature at 280°C for 8 minutes. |
Methods Filename: | GC_Method_Tissue.pdf |
Instrument Name: | Thermo Trace 1310 |
Column Name: | Thermo TraceGold TG-5SilMS |
Chromatography Type: | GC |
MS:
MS ID: | MS003361 |
Analysis ID: | AN003607 |
Instrument Name: | Thermo ISQ |
Instrument Type: | Single quadrupole |
MS Type: | EI |
MS Comments: | The MS was operated from 3.90 to 21.00 minutes in EI mode (-70eV) using select ion monitoring (SIM). Data were QC normalized using NOREVA (PMID: 28525573). |
Ion Mode: | POSITIVE |