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MB Sample ID: SA035017
Local Sample ID: | CD06241621 |
Subject ID: | SU000644 |
Subject Type: | Photosynthetic organism |
Subject Species: | Chlamydomonas reinhardtii |
Taxonomy ID: | 3055 |
Genotype Strain: | CC125 Wild type |
Species Group: | Microorganism |
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Sample Preparation:
Sampleprep ID: | SP000651 |
Sampleprep Summary: | Triplicate cultures were grown and treated as indicated and harvested at 72 h. The harvested cells were freeze-dried and weighted 10 ± 0.5 mg dry powder. To extract the metabolites, 500 uL of 75% MeOH in 0.15 M NaCl was added and metabolites were extracted using bead beating 50 Hz for 5 min. The homogenate was sonicated in water bath for 5 min and then 300 uL of 0.15 M NaCl was added and then 1 mL of chloroform:methanol (1:2) with 0.01% BHT was added. Samples were homogenized and allowed to phase separate on ice for 30 mins. Finally tubes were centrifuged for 10 min to separate the polar and non-polar layer. Top polar layer was removed and an aliquot of 200 uL was used for analysis. Samples were evaporated to dryness in SpeedVac and metabolites were reconstituted in 0.03% formic acid in analytical-grade water. Debris was removed via centrifugation and the supernatant was subjected to UPLC-MS analysis |
Extraction Method: | Bead beating |