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MB Sample ID: SA000012
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Sample Preparation:
| Sampleprep ID: | SP000001 |
| Sampleprep Summary: | - |
| Processing Storage Conditions: | Frozen tissues were kept in 2 ml round-bottomed Eppendorf tubes equipped with one 3 mm diameter steel ball, and homogenized using a Retsch (http://www.retch-us.com) ball mill for 30 sec at 25/sec |
| Extraction Method: | Ground tissue powder was kept in liquid nitrogen between homogenization and extraction. The extraction solvent was prepared by mixing isopropanol/ acetonitrile/water at the volume ratio 3:3:2 and degassing this mixture by directing a gentle stream of nitrogen through the solvent for 5 min. The solvent was cooled to )20 C prior to extraction. Randomly processing all samples of the study, 1 ml of cold solvent per 20 mg of ground tissue was added, vortexed for 10 sec, and shaken at 4 C for 5 min to extract metabolites and simultaneously precipitate proteins. After centrifugation at 12 800 g for 2 min, 90% of the supernatant was removed, taking are not to remove any residues from the pellet |
| Extract Concentration Dilution: | The supernatant was separated into two equal aliquots and concentrated to dryness in a Centrivap cold trap vacuum concentrator (http://www. labconco.com) at room temperature for 4 h |
| Extract Cleanup: | In order to fractionate complex lipids and waxes, the residue was re-suspended in 500 ll 50% aqueous acetonitrile and centrifuged at 12 800 g for 2 min. The supernatant was transferred to a 1.5 ml Eppendorf tube and concentrated to dryness in a vacuum concentrator |
| Extract Storage: | Dried extracts can be kept under nitrogen at -80 C for up to 4 weeks. In the study presented here, extracts were immediately derivatized for GCTOF mass spectrometry |
| Organ Specification: | Rosette leaf |
| Cell Type: | Arial portion |
