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MB Sample ID: SA052409
Local Sample ID: | Acorn1 |
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:
Subject:
Subject ID: | SU000926 |
Subject Type: | Plant |
Subject Species: | Quercus ilex |
Taxonomy ID: | 58334 |
Species Group: | Plant |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Acorn1 | SA052409 | FL009969 | Acorn | Sample Type |
Collection:
Collection ID: | CO000920 |
Collection Summary: | Germinated embryo, leaves and roots from four-months plantlets were collected separately, weighted and individually frozen in liquid nitrogen. |
Sample Type: | Plant |
Treatment:
Treatment ID: | TR000940 |
Treatment Summary: | Mature acorns from Holm oak (Quercus ilex L. subsp. ballota [Desf.] Samp.) were collected from a tree located in Aldea de Cuenca (province of Córdoba, Andalusia, Spain). Acorns were germinated and seedlings grew in a chamber under controlled conditions (a 12h photoperiod, a temperature of 21± 1ºC, a relative humidity of 60 ± 5% and an irradiance of 200 µmol m-2 s-1, Echevarría-Zomeño et al., 2009). |
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). |
Combined analysis:
Analysis ID | AN001451 |
---|---|
Analysis type | MS |
Chromatography type | GC |
Chromatography system | Agilent |
Column | Agilent HP5-MS (30m × 0.25mm, 0.25 um) |
MS Type | EI |
MS instrument type | Single quadrupole |
MS instrument name | Agilent 5975B |
Ion Mode | POSITIVE |
Units | area |
Chromatography:
Chromatography ID: | CH001020 |
Instrument Name: | Agilent |
Column Name: | Agilent HP5-MS (30m × 0.25mm, 0.25 um) |
Chromatography Type: | GC |
MS:
MS ID: | MS001341 |
Analysis ID: | AN001451 |
Instrument Name: | Agilent 5975B |
Instrument Type: | Single quadrupole |
MS Type: | EI |
Ion Mode: | POSITIVE |