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MB Sample ID: SA200923
| Local Sample ID: | b5_aos_179 |
| Subject ID: | SU002176 |
| Subject Type: | Invertebrate |
| Subject Species: | Caenorhabditis elegans |
| Taxonomy ID: | 6239 |
| Gender: | Hermaphrodite |
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Sample Preparation:
| Sampleprep ID: | SP002182 |
| Sampleprep Summary: | Frozen aliquots of 200,000 C. elegans worms were retrieved from -80°C and lyophilized in a VirTis® BenchTop™ “K” Series Freeze Dryer (SP Industries, Inc.). After lyophilization, each aliquot was weighed and stored at -80°C until homogenization. Aliquots were homogenized for three minutes in a Qiagen Tissuelyser 2, using glass beads. Homogenized worms were extracted with 1.5 mL of isopropanol (IPA) at -20°C overnight (approximately 12 hours), then pelleted and the supernatant transferred to separate 2 mL centrifuge tubes. Supernatants were then dried down to completion in a Labconco Centrivap and stored at -80°C for non-polar LC-MS analysis. The pellet was extracted a second time using 80:20 methanol:water (MeOH:H2O) (v:v) for 20 minutes at room temperature (RT) while shaking at 1500 rpm. Samples were again pelleted to separate proteins, and the supernatant was transferred to separate 2 mL centrifuge tubes, dried down to completion, and stored at -80°C for polar LC-MS analysis |
| Sampleprep Protocol Filename: | lcms_sequential_extraction_method.pdf |
| Processing Storage Conditions: | -80℃ |
| Extraction Method: | IPA for non-polar; 80/20 MeOH/H2O for polar |
| Extract Storage: | -80℃ |
| Sample Resuspension: | 75 µL of IPA containing isotopically labeled lipid standards; 75 µL of 80:20 MeOH: H2O containing isotopically labeled arginine, hypoxanthine, hippuric acid, and methionine |
