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MB Sample ID: SA175088
| Local Sample ID: | Copepodamide 8 |
| Subject ID: | SU001960 |
| Subject Type: | Other organism |
| Subject Species: | Alexandrium minutum |
| Taxonomy ID: | 39455 |
| Genotype Strain: | CCMP 113 |
| Gender: | Not applicable |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001960 |
| Subject Type: | Other organism |
| Subject Species: | Alexandrium minutum |
| Taxonomy ID: | 39455 |
| Genotype Strain: | CCMP 113 |
| Gender: | Not applicable |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Copepodamide 8 | SA175088 | FL021508 | Copepodamide | Treatment |
Collection:
| Collection ID: | CO001953 |
| Collection Summary: | Alexandrium minutum cells were collected by vacuum filtration onto GF/F filters and quenched with liquid nitrogen. Frozen cells with filters were stored in foil (previously muffled for 3 h at 450 °C) at -80 °C until extraction. |
| Collection Protocol Filename: | Predator_Cues_Target_Signaling_Pathways_in_Toxic_Algal_Metabolome_protocol.pdf |
| Sample Type: | Algae |
| Collection Method: | Filtration and Freeze |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001972 |
| Treatment Summary: | Twice, first at the start of the experiment and again 24 h later, 1.5 mL of either 0.2 µM copepodamides (copepod cue) in DMSO (treatment, n=20) or DMSO alone (control, n=20) were added aseptically to 300 mL cultures of Alexandrium minutum resulting in a final concentration of 1 nM copepodamides. |
| Treatment Protocol Filename: | Predator_Cues_Target_Signaling_Pathways_in_Toxic_Algal_Metabolome_protocol.pdf |
Sample Preparation:
| Sampleprep ID: | SP001966 |
| Sampleprep Summary: | Polar extracts were dissolved at a concentration equivalent to 1.70 x 10^7 A. minutum cells mL-1 in 90:10 H2O/D2O (99.96% atom D2O; Cambridge Isotope Labs) with 0.2 mM phosphate buffer (pH 7.4) and 0.25 mM 3 (trimethylsilyl)propionic 2,2,3,3d 4 acid (TMSP), as internal standard, for 1H NMR spectroscopy analysis. The media blank extract was prepared in the smallest volume of solvent possible (175 µL). Particulates were excluded by briefly centrifuging the samples and then carefully transferring the extracts into 3 mm NMR tubes. The extracts were analyzed using a Bruker Avance IIIHD 800 MHz NMR spectrometer equipped with a 3 mm triple resonance broadband cryoprobe. Spectra for the polar extracts were acquired using a 1D water presaturation (Bruker zgpr pulse sequence) with a spectral width of 12 kHz centered at 3758.36 Hz, with a relaxation delay 3 s, over 480 scans. |
| Sampleprep Protocol Filename: | Predator Cues Target Signaling Pathways in Toxic Algal Metabolome_protocol.pdf |
| Extract Storage: | -80℃ |
Analysis:
| MB Sample ID: | SA175088 |
| Analysis ID: | AN003046 |
| Analysis Type: | NMR |
| Results File: | Preprocessed_binned_polar_NMRmetabolomics.txt |
| Units: | area under the curve |
NMR:
| NMR ID: | NM000211 |
| Analysis ID: | AN003046 |
| Instrument Name: | Bruker Avance IIIHD |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 1D-1H |
| Spectrometer Frequency: | 800 MHz |
| NMR Probe: | 3 mm triple resonance broadband cryoprobe |
| NMR Solvent: | 90:10 H2O/D2O (99.96% atom D2O; Cambridge Isotope Labs) with 0.2 mM phosphate buffer (pH 7.4) and 0.25 mM 3-(trimethylsilyl)propionic-2,2,3,3d4-acid |
| NMR Tube Size: | 3 mm |
| Pulse Sequence: | zgpr |
| Number Of Scans: | 480 |
| Relaxation Delay: | 3s |
| Spectral Width: | 12 kHz |
| Baseline Correction Method: | Spline |
| Binned Increment: | 0.005 ppm |
