Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA216951

Local Sample ID:	sample29
Subject ID:	SU002338
Subject Type:	Plant
Subject Species:	Arabidopsis thaliana
Taxonomy ID:	3702
Genotype Strain:	WT Col-0, atg7-2 (GABI_655B06), atg9-4 (SALK_145980)
Age Or Age Range:	11-day old
Gender:	Not applicable

Select appropriate tab below to view additional metadata details:

Sample Preparation:

Sampleprep ID:	SP002344
Sampleprep Summary:	Lipids were extracted using a modification of a standard protocol. The collected plant tissues (leaf or root) were transferred in to a 50 mL Teflon-lined screw-caped glass tube (Thermo Fisher Scientific) containing 3 mL preheated isopropanol (Thermo Fisher Scientific) containing 0.01 % (v/v) butylated hydroxytoluene (BHT) (MilliporeSigma) and 1 µM 1,2-didecanoyl-sn-glycero-3-phosphocholine (PC 20:0)(MilliporeSigma) as an internal standard. The tubes were incubated at 75 °C for 15 min to quench the action of any lipases. Following the addition of 1.5 mL chloroform and 0.6 mL water the mixture was vigorously shaken at room temperature for 1 h. The clear liquid extract was transferred to another 50 mL tube using glass Pasteur pipettes, and the remnant tissue was further extracted for 30-minutes with another 4 mL chloroform/methanol (2:1) that contained 0.01% BHT. The clear liquid from this second extraction was removed and combined with the initial liquid extract. This chloroform/methanol (2:1) extraction was repeated three times, and the last extraction being incubated overnight. The residue tissue remaining after lipid extraction was dried at 105 °C, and the dry weight of each sample determined; each leaf tissue sample weighed approximately 20 mg, and each root tissue sampled weighed approximately 10 mg. All extract aliquots from each biological sample were combined into a single screw capped tube and stored at -80 °C under a nitrogen gas atmosphere. The solvent from each extract removed by evaporation with the aid of a stream of N2 gas, and the lipid residue was dissolved in 1 mL chloroform and transferred to 2.0 mL clear glass vial with Teflon-lined screw cap (Thermo Fisher Scientific). The solvent was again evaporated with N2 gas, and the vials were shipped, overnight on dry ice to Kansas Lipidomics Research Center (https://www.k-state.edu/lipid/) for lipidomics analysis.

Sampleprep ID:

SP002344

Sampleprep Summary:

Lipids were extracted using a modification of a standard protocol. The collected plant tissues (leaf or root) were transferred in to a 50 mL Teflon-lined screw-caped glass tube (Thermo Fisher Scientific) containing 3 mL preheated isopropanol (Thermo Fisher Scientific) containing 0.01 % (v/v) butylated hydroxytoluene (BHT) (MilliporeSigma) and 1 µM 1,2-didecanoyl-sn-glycero-3-phosphocholine (PC 20:0)(MilliporeSigma) as an internal standard. The tubes were incubated at 75 °C for 15 min to quench the action of any lipases. Following the addition of 1.5 mL chloroform and 0.6 mL water the mixture was vigorously shaken at room temperature for 1 h. The clear liquid extract was transferred to another 50 mL tube using glass Pasteur pipettes, and the remnant tissue was further extracted for 30-minutes with another 4 mL chloroform/methanol (2:1) that contained 0.01% BHT. The clear liquid from this second extraction was removed and combined with the initial liquid extract. This chloroform/methanol (2:1) extraction was repeated three times, and the last extraction being incubated overnight. The residue tissue remaining after lipid extraction was dried at 105 °C, and the dry weight of each sample determined; each leaf tissue sample weighed approximately 20 mg, and each root tissue sampled weighed approximately 10 mg. All extract aliquots from each biological sample were combined into a single screw capped tube and stored at -80 °C under a nitrogen gas atmosphere. The solvent from each extract removed by evaporation with the aid of a stream of N2 gas, and the lipid residue was dissolved in 1 mL chloroform and transferred to 2.0 mL clear glass vial with Teflon-lined screw cap (Thermo Fisher Scientific). The solvent was again evaporated with N2 gas, and the vials were shipped, overnight on dry ice to Kansas Lipidomics Research Center (https://www.k-state.edu/lipid/) for lipidomics analysis.