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MB Sample ID: SA080595
| Local Sample ID: | M12D3 |
| Subject ID: | SU001230 |
| Subject Type: | Invertebrate |
| Subject Species: | Metacarcinus magister |
| Taxonomy ID: | 29965 |
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Sample Preparation:
| Sampleprep ID: | SP001238 |
| Sampleprep Summary: | 15mg of whole crab was placed into a 1.5mL ependorph tube. 2 x 3mm grinding beads were added to each sample. 225 µL of cold MeOH with quality controls was added to each samples. Batches of samples were ground with GenoGrinder for 30 seconds at 1500 rpm. 750µL of methyl tert-butyl Ether (MTBE) was added to each sample. Samples were vortexed for 10 seconds and then shaken at 4°C for 5 minutes using an Orbital Mixer. 188 uL of LC-MS grade water was added to each sample. Vortex for 10 seconds and then centrifuged for 2 minutes at 14,000 rcf. 2 x 350µL aliquots were removed from the top, organic layer, one submitted for analysis and the other stored as backup in -20°C. 2 x 125µL aliquots were removed from the bottom, polar layer, one submitted for analysis, the other stored at -20°C for backup. |
| Sample Resuspension: | After drying, samples were resuspended in 110 µL of 9:1 methanol:toulene with 50 nM CUDA as an internal standard. Samples were vortexed for 10 seconds, then sonicated in room temperature water for 5 minutes, then centrifuged at 16,100 rcf for 2 minutes. 50 µL of the supernatant was removed and placed in an amber vial for lipidomics analysis. |
