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MB Sample ID: SA213986
| Local Sample ID: | 16_Cotyledon_2-neg |
| Subject ID: | SU002325 |
| Subject Type: | Plant |
| Subject Species: | Camelina Sativa |
| Age Or Age Range: | 10 days after fertilization |
| Species Group: | Developing seeds |
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Sample Preparation:
| Sampleprep ID: | SP002331 |
| Sampleprep Summary: | Frozen cotyledon samples from Camelina were homogenized using a tissue lyser and the extraction of lipids was carried out using a phase separation method previously described26. Briefly, 1 mL 7:3 methanol:chloroform (-20°C) containing the ultimateSPLASHTM ONE lipid mix (Avanti Polar lipids, Alabaster, AL) as internal standard (1:20 dilution) was added to the samples, vortexed vigorously and incubated on a rotary shaker for 2 hours at 4°C. After incubation, 500 µL of ice-cold water was added to the samples, vortexed and centrifuged at 14,000 rpm at 4°C for 10 min to achieve phase separation. The upper aqueous phase was carefully removed, 200 µL of methanol was added to the remaining organic phase containing lipids and centrifuged at 14,000 rpm for 5 min to pellet the debris. The organic phase (supernatant) was transferred to a glass tube and dried using a speedvac centrifuge. Samples were re-suspended in 200 µL of 49:49:2 acetonitrile: methanol: chloroform, filtered using 0.2 µm PTFE microcentrifuge filters and transferred to a glass vial for RPLC-HRMS analysis. |
| Processing Storage Conditions: | -80℃ |
| Extraction Method: | methanol:chloroform:water |
