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MB Sample ID: SA314731

Local Sample ID:Hypoxia_1_1_neg
Subject ID:SU002993
Subject Type:Plant
Subject Species:Lycopersicon esculentum

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Sample Preparation:

Sampleprep ID:SP002999
Sampleprep Summary:Lipids from tomato roots were extracted with a modified procedure adapted from Shiva and colleagues [20]. In brief, 0.1 g of grinded roots (cooled on liquid nitrogen) were added to a glass reaction tube filled with isopropanol supplemented with 0.01 % BHT. The mixture was incubated for 15 min at 75 °C and cooled on ice afterward. To the cooled-down mixture, 0.5 volume of chloroform, 0.2 volume of water and 3 µL of EquiSPLASH Lipidomix were added. Lipids were extracted for 1 h on ice with occasional vortexing in between. Reaction tubes were centrifuged at 5000 x g to induce phase separation, and the organic phase was transferred to another glass reaction tube. To the remaining water phase, 1.33 volumes of a chloroform:methanol mixture (2:1, v/v) supplemented with 0.1 % BHT was added and incubated for 30 min on ice with occasional vortexing. Reaction tubes were again centrifuged at 5000 rpm, and the chloroform phase was transferred to the second glass vial. The chloroform:methanol extraction step was repeated with the remaining water phase 3 times. Afterward, 0.33 volumes of KCl (1 M) were added to the combined lipid extracts and vortexed. The upper water phase was removed, and 0.66 volumes of water were added and vortexed. The upper phase was again removed, and sodium sulfate was added to remove the remaining water. Finally, the lipid extract was dried under constant N2-flow with a TurboVap sample evaporator (Biotage, Sweden) and frozen at -80°C until MS analysis.
Processing Storage Conditions:On ice
Extract Storage:-80℃
Sample Resuspension:chloroform:methanol:isopropanol (1:2:4 v/v/v, supplemented with 5 mM ammonium formate)
Sample Derivatization:none
Sample Spiking:EquiSPLASH
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