General Information info
Experiment Description info
|Strain||C57BLKS/J-m+/+db and mutants|
|Data type||metabolites at steady-state|
|Execution date||not specified|
Experimental Details info
|Dilution rate (h-1)||not specified|
|Working volume (L)||0.0005|
|Biomass concentration (g/L)||not specified|
|General protocol information||
The mice were sacrificed by decapitation at 17-wk of age, and specimens of hippocampus were collected manually.
Quenching procedure: specimens of hippocampus were dissected immediately, snap-frozen in liquid nitrogen and stored at -80 °C until use.
Extraction technique: chloroform
Sample analyzing method: NMR
|Methods description - Notes||
The preparation of the brain extracts was based on the previous reference . The frozen tissue was weighed and ground using an electric homogenizer with ice-cold methanol (4 mL/g) and distilled water (0.85 mL/g) at 4°C and the mixture was vortexed. Chloroform (2 mL/g) and distilled water (2 mL/g) was added and mixed again. After keeping on ice for 15 min, the homogenate was centrifuged at 1,000 g for 15 min at 4°C. The supernatant was extracted and lyophilized for about 24h. The metabolite mixture obtained was then weighed and dissolved in 0.6 mL of 99.5% D2O for NMR spectroscopy. One-dimensional ZGPR pulse sequence was used, and the main parameters were set as follows: data points, 64 K; spectral width, 12,000 Hz; relaxation delay, 10 s. The spectral segments for each NMR spectrum were normalized to the total sum of the spectral intensity to partially compensate for differences in concentration of the many metabolites in the samples .
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