Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA052416

Local Sample ID:	Root2
Subject ID:	SU000926
Subject Type:	Plant
Subject Species:	Quercus ilex
Taxonomy ID:	58334
Species Group:	Plant

Select appropriate tab below to view additional metadata details:

Sample Preparation:

Sampleprep ID:	SP000933
Sampleprep Summary:	Metabolites were extracted from each type of organs under study (leaves, roots and acorns), as described by (Valledor et al., 2014). 600 µL of cold (4oC) metabolite extraction buffer (methanol: chloroform: water; 5:2:2) were added to 15 mg of dried tissue (frozen, lyophilized weight). Powdered samples were extracted by vortexing for 10 s and sonicating in an ultrasonic bath at 4oC and maximum frequency (40 kHz). Samples were centrifuged at 20.000 g for 4 min at 4oC and the supernatant were transferred to 2 mL microcentrifuge tubes that contained 400 µL of phase separation mix (chloroform: water; 1:1). Tubes with metabolites were centrifuged at 20.000 g for 4 min at 4oC. The two phases were clearly defined with a sharp interface. Polar and non-polar metabolites, upper and lower layers respectively were transferred to new tubes. These two fractions were washed again (200 µL of cold (4oC) chloroform for polar layer and with 200 µL of cold (4oC) water for non-polar layer), centrifuged, and fractioned again. Polar and non-polar layers were saved to new tubes and dried completely with a micro concentrator Speedvac (Eppendorf Vacuum Concentrator Plus/5301).

Sampleprep ID:

SP000933

Sampleprep Summary:

Metabolites were extracted from each type of organs under study (leaves, roots and acorns), as described by (Valledor et al., 2014). 600 µL of cold (4oC) metabolite extraction buffer (methanol: chloroform: water; 5:2:2) were added to 15 mg of dried tissue (frozen, lyophilized weight). Powdered samples were extracted by vortexing for 10 s and sonicating in an ultrasonic bath at 4oC and maximum frequency (40 kHz). Samples were centrifuged at 20.000 g for 4 min at 4oC and the supernatant were transferred to 2 mL microcentrifuge tubes that contained 400 µL of phase separation mix (chloroform: water; 1:1). Tubes with metabolites were centrifuged at 20.000 g for 4 min at 4oC. The two phases were clearly defined with a sharp interface. Polar and non-polar metabolites, upper and lower layers respectively were transferred to new tubes. These two fractions were washed again (200 µL of cold (4oC) chloroform for polar layer and with 200 µL of cold (4oC) water for non-polar layer), centrifuged, and fractioned again. Polar and non-polar layers were saved to new tubes and dried completely with a micro concentrator Speedvac (Eppendorf Vacuum Concentrator Plus/5301).