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MB Sample ID: SA210642
| Local Sample ID: | T3.30.LWS2_pos |
| Subject ID: | SU002284 |
| Subject Type: | Plant |
| Subject Species: | Pinus pinaster |
| Taxonomy ID: | 71647 |
| Age Or Age Range: | one-two years |
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Sample Preparation:
| Sampleprep ID: | SP002290 |
| Sampleprep Summary: | Metabolites were extracted from 20 mg (lyophilized weight) of needles. Metabolite extraction was performed according to Valledor et al., (2014). Briefly, 600 µL of cold (4 ºC) metabolite extraction solution (methanol: chloroform: H2O (2.5:1:0.5) was added to each tube and strongly vortexed. Then, the tubes were incubated in a cold ultrasound bath for 10 min. Later, the tubes were centrifuged at 20.000 x g for 6 min at 4 °C. The supernatant containing metabolites from each tube was transferred to a new tube containing 300 µL of chloroform: water (1:1) to allow phase separation. Six hundred µL of cold (4 ºC) metabolite extraction solution were added to the remaining pellets, and vortexing, ultrasound bath, and centrifugation were repeated. The new supernatant was transferred to the previous tube that contained the phase separation solution and the old supernatant. These tubes were vortexed and then centrifuged at 15.000 x g for 5 min at 4 °C. After centrifugation, two layers are formed; the upper-aqueous layer (methanol: water) containing the polar metabolites was transferred to a separate microcentrifuge tube and then cleaned from non-polar metabolites adding 300 µL of cold (4 ºC) chloroform: water (1:1), vortexed, and centrifuged at 15.000 x g for 4 min at 4 °C. The new upper phase was transferred to a new tube. The polar extract was dried using a speedvac at 25 °C. |
| Processing Method: | Methanol:Chloroform:Water |
| Processing Storage Conditions: | On ice |
| Extraction Method: | Methanol:Chloroform:Water |
| Extract Enrichment: | Polar metabolites |
| Extract Cleanup: | Centrifugation |
| Extract Storage: | -80℃ |
| Sample Resuspension: | Methanol |
| Sample Derivatization: | NO |
| Sample Spiking: | NO |
