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MB Sample ID: SA035030
Local Sample ID: | 461_A_3 |
Subject ID: | SU000645 |
Subject Type: | Photosynthetic organism |
Subject Species: | Chlamydomonas reinhardtii |
Taxonomy ID: | 3055 |
Genotype Strain: | Wild Type |
Species Group: | Microorganism |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000645 |
Subject Type: | Photosynthetic organism |
Subject Species: | Chlamydomonas reinhardtii |
Taxonomy ID: | 3055 |
Genotype Strain: | Wild Type |
Species Group: | Microorganism |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
461_A_3 | SA035030 | FL007488 | WD10461 | Class |
Collection:
Collection ID: | CO000639 |
Collection Summary: | Cells were pre-grown to mid-log phase and treated with 5 selected compounds (final concentration 5 µM) with an initial cell density of 1.0 x 106 cells/mL (100 mL volume; in triplicate) and allowed to grow for 72 h. After 72 h of growth, cells were harvested, media removed and freeze-dried. Accurately measured 50 ± 0.5 mg of freeze dried powder was used for metabolite extraction. Sample powder was pulverized with a single steel ball using TissueLyser LT (Qiagen) at 50 Hz speed for 5 min |
Sample Type: | Algae |
Treatment:
Treatment ID: | TR000659 |
Treatment Summary: | Cells were treated either with compounds (5 uM) final concentration in DMSO or DMSO alone (in case of control). Mid-log phase cells were used as starter culture at a initial inoculum density of 1.0E06 cells/mL in 250 mL flasks with 100 mL of TAP media. Cells were allowed to grow in orbital shaker under constant light for 72 hours. After 72 hours, experiment was terminated and cells were harvested via centrifugation. |
Sample Preparation:
Sampleprep ID: | SP000652 |
Sampleprep Summary: | Harvested cells were flash frozen in liquid N and freeze dried. Accurately measured 50 ± 0.5 mg of freeze dried powder was used for metabolite extraction. Sample powder was pulverized with a single steel ball using TissueLyser LT (Qiagen) at 50 Hz speed for 5 min. One milliliter of extraction buffer containing MeOH:CHCl3:H2O (5:2:2; v/v/v; pre-cooled at -20 °C) was added and vortexed for 5 min. Ribitol (0.2 mg/mL in water; 10 µL) was spiked in the extraction buffer as internal standard in order to identify potential chromatographic errors. The homogenized material was centrifuged at 14000 rpm for 5 min and the supernatant was transferred to new tubes. 400 µL of pure water was added to the supernatant, samples were vortexed and centrifuged at 14,000 rpm for 5 min. The upper polar phase was transferred to new tubes for GC-MS analysis. An aliquot of 300 µL was dried out in vacuum concentrator without heating. To the dried material, 10 µL methoxyamine HCL in 100% pyridine (40 mg/mL) was added and shaken at 30 °C for 90 minutes and subsequently 90 µL of MSTFA 1% TMCS was added for trimethylsilylation of acidic protons and shaken at 37 °C for 30 minutes. The reaction mixture was transferred to GCvials with glass microinserts and closed by crimp caps. GC-MS data acquisition was performed as per previously published report (Wase et al., 2014) |
Combined analysis:
Analysis ID | AN000954 |
---|---|
Analysis type | MS |
Chromatography type | GC |
Chromatography system | Agilent 6890N |
Column | Agilent DB-5MS UI Capillary column |
MS Type | EI |
MS instrument type | Single quadrupole |
MS instrument name | Agilent 5973 |
Ion Mode | POSITIVE |
Units | peak area |
Chromatography:
Chromatography ID: | CH000679 |
Instrument Name: | Agilent 6890N |
Column Name: | Agilent DB-5MS UI Capillary column |
Internal Standard: | Ribitol |
Sample Injection: | 1 uL |
Chromatography Type: | GC |
MS:
MS ID: | MS000849 |
Analysis ID: | AN000954 |
Instrument Name: | Agilent 5973 |
Instrument Type: | Single quadrupole |
MS Type: | EI |
Ion Mode: | POSITIVE |