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MB Sample ID: SA019317
| Local Sample ID: | 140529bmssa42_1 |
| Subject ID: | SU000423 |
| Subject Type: | Cells |
| Subject Species: | Chlorella minutissima |
| Taxonomy ID: | 3081 |
| Species Group: | Microorganism |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000423 |
| Subject Type: | Cells |
| Subject Species: | Chlorella minutissima |
| Taxonomy ID: | 3081 |
| Species Group: | Microorganism |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 140529bmssa42_1 | SA019317 | FL006534 | 10 g/L | Glucose conc |
Collection:
| Collection ID: | CO000417 |
| Collection Summary: | Samples were collected daily from each culture over 5 days (starting at day = 0) |
| Sample Type: | Cells |
Treatment:
| Treatment ID: | TR000437 |
| Treatment Summary: | Cultures #1-6 were supplied with 10 g/L glucose, cultures #7-12 were not supplied with glucose (autotrophic) |
| Treatment Protocol Filename: | Objective_2_Exp_6_Mixo_Metabolomics.xlsx |
| Treatment Protocol Comments: | Cultures were grown with 125 ml/min air, 10,000 lux illumination |
Sample Preparation:
| Sampleprep ID: | SP000430 |
| Sampleprep Summary: | Quenching 1. Add 5mL of quenching solvent to each 15mL tube, and let sit for at least 30 minutes in ice bath to get to temperature. 2. Add 1mL of cell culture (OD600=1.5) to tube, cap, and gently invert several times. 3. Centrifuge 1,000xg for 5 minutes. 4. Remove supernatant, and store supernatant and cell pellet in -80C until further use. Extraction 1. Add 0.5mL of extraction solvent to tube, gently pipet to remove all cells, transfer cells to 2mL eppendorf tube. Repeat for a total of 1mL extraction solvent + cells in 2mL eppendorf tube. 2. Add 2 small stainless steel grinding beads to eppendorf tube 3. Use the GenoGrinder to grind for 3 minutes at 1,250 rpm. 4. Centrifuge at 14,000xg for 5 minutes. 5. Transfer supernatant to a fresh 2mL eppendorf tube. 6. Add 1mL of extraction solvent to tube containing cell pellet + beads, and repeat steps 3 and 4. 7. Collect supernatant, and combine with supernatant collected in step 5. Total volume of extracted sample will be approximately 2mL. 8. Dry down 50uL of extracted sample in 1.5mL eppendorf tube for GC-TOF analysis. 9. Store backups in -20 or -80C. -20°C bath 5mL quenching solvent 1mL cells+broth |
| Sampleprep Protocol Filename: | SOP_Extraction_of_Yeast_Cells.pdf |
Combined analysis:
| Analysis ID | AN000641 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 6890N |
| Column | Restek Corporation Rtx-5Sil MS |
| MS Type | EI |
| MS instrument type | GC Ion Trap |
| MS instrument name | Varian 210-MS GC Ion Trap |
| Ion Mode | POSITIVE |
| Units | counts |
Chromatography:
| Chromatography ID: | CH000466 |
| Methods Filename: | Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf |
| Instrument Name: | Agilent 6890N |
| Column Name: | Restek Corporation Rtx-5Sil MS |
| Column Pressure: | 7.7 PSI |
| Column Temperature: | 50-330C |
| Flow Rate: | 1 ml/min |
| Injection Temperature: | 50 C ramped to 250 C by 12 C/s |
| Sample Injection: | 0.5 uL |
| Oven Temperature: | 50°C for 1 min, then ramped at 20°C/min to 330°C, held constant for 5 min |
| Transferline Temperature: | 230C |
| Washing Buffer: | Ethyl Acetate |
| Sample Loop Size: | 30 m length x 0.25 mm internal diameter |
| Randomization Order: | Excel generated |
| Chromatography Type: | GC |
MS:
| MS ID: | MS000573 |
| Analysis ID: | AN000641 |
| Instrument Name: | Varian 210-MS GC Ion Trap |
| Instrument Type: | GC Ion Trap |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
| Ion Source Temperature: | 250 C |
| Ionization Energy: | 70 eV |
| Mass Accuracy: | Nominal |
| Source Temperature: | 250 C |
| Scan Range Moverz: | 85-500 Da |
| Scanning Cycle: | 17 Hz |
| Scanning Range: | 85-500 Da |
| Skimmer Voltage: | 1850 V |
