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MB Sample ID: SA322004
| Local Sample ID: | 19 |
| Subject ID: | SU003070 |
| Subject Type: | Plant |
| Subject Species: | Thalassiosira pseudonana |
| Taxonomy ID: | 296543 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003070 |
| Subject Type: | Plant |
| Subject Species: | Thalassiosira pseudonana |
| Taxonomy ID: | 296543 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 19 | SA322004 | FL038422 | 20 | Temperature |
| 19 | SA322004 | FL038422 | 0 | Bacteria presence |
Collection:
| Collection ID: | CO003063 |
| Collection Summary: | Based on pre-experimental axenic growth curves, harvests occurred at days 3 (28°C), 4 (20°C), and 6 (14°C), starting 7 h into the light cycle. At each harvest, subsamples of 600 mL were filtered onto 2.0-µm Isopore filters (Millipore, Burlington, MA) for diatom endometabolite analysis and stored in 50-mL centrifuge tubes at -80°C. |
| Collection Protocol Filename: | 2_Collection protocol_UGA_temp_Oct2023_main.docx |
| Sample Type: | Algae |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR003079 |
| Treatment Summary: | Six treatment combinations of a marine diatom strain Thalassiosira pseudonana CCMP1335 were prepared: treatments incubated axenically at either 14, 20, or 28 oC, and treatments co-cultured with a bacterial strain Ruegeria pomeroyi DSS-3 at the corresponding temperatures (four replicates for each). L1 media was used with NaH13CO3 as a source of bicarbonate and a salinity of 35 ppt. The diatom used for the co-cultured treatments was B12 stressed to emphasize the known co-existing system. The light cycle consisted of 16 h light (120 µmol photons m-2 s-1) and 8 h of dark. |
| Treatment Protocol Filename: | 3_Treatment protocol_UGA_temp_Oct2023_main.docx |
Sample Preparation:
| Sampleprep ID: | SP003076 |
| Sampleprep Summary: | Filter samples were transferred into 15-mL of ultrapure MilliQ water in 50-mL tubes, and diatom cells were removed from the filters by sonication in an ice-water bath for 7 min (cycle: 50 s on and 10 s off). The liquid fraction was subsequently collected in new tubes and the procedure repeated three times, after which fractions were combined and stored at -80°C until further processing. Samples were lyophilized (Labconco, Kansas City, MO, USA) and pellets mixed with 600 μL of phosphate buffer (30 mmol L-1 phosphate in deuterated water, pH 7.4) and 1 mmol L-1 internal standard 2,2-dimethyl-2-silapentane-5-sulfonate (DSS). Samples were vortexed for 5 min, centrifuged at 20,800 relative centrifugal force (RCF) for 10 min, and supernatants were transferred to 5-mm NMR tubes (Bruker, Billerica, MA, USA). Extraction and buffer blank controls were also prepared. Additionally, one pooled control sample was prepared by combining aliquots of all the samples and used for annotation. |
| Sampleprep Protocol Filename: | 4_Sample preparation protocol_UGA_temp_Oct2023_main.docx |
Analysis:
| MB Sample ID: | SA322004 |
| Analysis ID: | AN004857 |
| Analysis Type: | NMR |
| Analysis Protocol File: | 5_Analysis protocol_UGA_temp_Oct2023_main.docx |
| Software Version: | TopSpin version 3.5 |
| Num Factors: | 6 |
| Num Metabolites: | 16 |
| Units: | Intensity |
NMR:
| NMR ID: | NM000269 |
| Analysis ID: | AN004857 |
| Instrument Name: | AVANCE III HD instrument (Bruker) |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 2D-1H-13C |
| Spectrometer Frequency: | 600 MHz |
| NMR Probe: | TCI cryoprobe |
| NMR Tube Size: | 5-mm |
