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MB Sample ID: SA212537
| Local Sample ID: | 37897_H |
| Subject ID: | SU002315 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL6 and FVB mixed strain |
| Age Or Age Range: | 54-59 weeks old |
| Gender: | Male and female |
| Animal Feed: | Specialty Feeds Irradiated Rat and Mouse Standard Chow Diet |
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Sample Preparation:
| Sampleprep ID: | SP002321 |
| Sampleprep Summary: | tissues was homogenised in 1xPBS and then sonicated with a probe-sonicator for 15 seconds, 23 amplitude. BCA assays were then conducted to determine protein concentrations of these homogenates. Lipid extraction was conducted using 10ul of sample (ventricle, skeletal muscle homogenate at 5mg/ml, fat homogenate at 2mg/ml and liver homogenate at 2.5mg/ml) using the single phase chloroform methanol method. 10ul of internal standards and 200ul of chloroform:methanol (1:2) were added to samples before the mixture was vortexed. Samples were then placed on a rotary shaker for 10 mins at a speed of 90 before being transferred to a bath sonicator. Samples were then sonicated for 30 mins at water temperature below 28 degrees. Samples were then removed and rested at room temperature for 20 mins. Samples were then centrifuged at 13000rpm for 10 minutes. 200ul of the supernatant was then transferred to 0.5ml polypropylene 96 well plates, and spun dried using a speedvac vacuum concentrator. Lipids were reconstituted in 50ul water saturated butanol + 50ul of Ammonium Formate. |
| Sampleprep Protocol Filename: | Agilent_appnote |
| Extract Storage: | -80℃ |
