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MB Sample ID: SA211367

Local Sample ID:S4
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

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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 IDMB Sample IDFactor Level IDLevel ValueFactor Name
S4SA211367FL025774KOGenotype

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
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