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MB Sample ID: SA005591
| Local Sample ID: | 338 |
| Subject ID: | SU000121 |
| Subject Type: | Insect |
| Subject Species: | Drosophila melanogaster |
| Taxonomy ID: | 7227 |
| Genotype Strain: | Originating from same base population as Drosophila melanogaster genetic reference panel, selected for fast (hardy) and slow (susceptible) recovery from chill coma. |
| Age Or Age Range: | 5-7 day-old adults (mated) |
| Weight Or Weight Range: | 0.17-0.77mg |
| Height Or Height Range: | NA |
| Gender: | Female |
| Animal Housing: | 25 degC |
| Animal Light Cycle: | 12L:12D |
| Animal Feed: | Standard cornmeal molasses medium |
| Animal Water: | in diet only |
| Species Group: | Insect |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000121 |
| Subject Type: | Insect |
| Subject Species: | Drosophila melanogaster |
| Taxonomy ID: | 7227 |
| Genotype Strain: | Originating from same base population as Drosophila melanogaster genetic reference panel, selected for fast (hardy) and slow (susceptible) recovery from chill coma. |
| Age Or Age Range: | 5-7 day-old adults (mated) |
| Weight Or Weight Range: | 0.17-0.77mg |
| Height Or Height Range: | NA |
| Gender: | Female |
| Animal Housing: | 25 degC |
| Animal Light Cycle: | 12L:12D |
| Animal Feed: | Standard cornmeal molasses medium |
| Animal Water: | in diet only |
| Species Group: | Insect |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 338 | SA005591 | FL001407 | after | time |
| 338 | SA005591 | FL001407 | susceptible | hardiness |
| 338 | SA005591 | FL001407 | 1 | population replicate |
Collection:
| Collection ID: | CO000104 |
| Collection Summary: | Flies are snap-frozen in liquid nitrogen, counted into pools of 10, and stored at -80 degC until extraction. |
| Collection Protocol Filename: | Fly_rearing_and_sample_collection_24Sept2014.docx |
| Sample Type: | whole bodies |
| Collection Method: | Snap-freeze in liquid nitrogen |
| Collection Location: | University of Florida |
| Storage Conditions: | -80 degC |
| Collection Vials: | 2mL eppendorf tubes |
| Storage Vials: | 2mL eppendorf tubes |
Treatment:
| Treatment ID: | TR000122 |
| Treatment Summary: | Flies are exposed to 3h in an ice slurry, then allowed to recover at room temperature for 5min. |
| Treatment Protocol Filename: | Fly_rearing_and_sample_collection_24Sept2014.docx |
| Treatment: | abiotic - cold exposure, 3h in ice slurry |
| Treatment Dose: | ice slurry |
| Treatment Doseduration: | 3h |
| Animal Endp Tissue Coll List: | 10 whole flies/sample |
Sample Preparation:
| Sampleprep ID: | SP000117 |
| Sampleprep Summary: | - |
| Sampleprep Protocol Filename: | sample_prep_protocol_24Sept2014.docx sample_spiking_protocol_24Sept2014.docx |
| Sampleprep Protocol Comments: | Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911-7. |
| Processing Method: | lyophilization, homogenization, solvent extraction |
| Processing Storage Conditions: | store at -20C |
| Extraction Method: | modified Bligh and Dyer protocol |
| Extract Concentration Dilution: | 2:2:1.8 (methanol:chloroform:water) |
| Extract Storage: | -20C |
| Sample Resuspension: | Samples were re-suspended in 620 ?l buffer (100 mM potassium phosphate [pH 7.3], 0.1% sodium azide, 1 mM trimethylsilylproprionate [TMSP] in D2O) |
| Sample Spiking: | 6 internal standards (proline, ?-alanine, tyrosine, histidine, phosphocholine, and 3-hydroxykynurenine) added: 6 ?L of standards were added to a concentration of 2x the Chenomx-determined concentration for each metabolite |
Analysis:
| MB Sample ID: | SA005591 |
| Analysis ID: | AN000170 |
| Laboratory Name: | Molecular Science Research Center (MSRC), Claflin University, Orangeburg, SC |
| Analysis Type: | NMR |
| Analysis Comments: | NMR: 2D 1H-13C HSQC |
| Acquisition Date: | Summer 2012 |
| Acquisition Parameters File: | nmr_data_acquisition_and_processing_protocol.doc |
| Instrument Parameters File: | Cold_adaptation_in_Drosophila_melanogaster.zip |
| Software Version: | TOPSPIN 2.1 |
| Operator Name: | Miki Watanabe and Arezue Boroujerdi |
| Chromatography ID: | CH000116 |
| Num Factors: | 12 |
| MB Sample ID: | SA005591 |
| Analysis ID: | AN000169 |
| Laboratory Name: | Molecular Science Research Center (MSRC), Claflin University, Orangeburg, SC |
| Analysis Type: | NMR |
| Analysis Comments: | NMR: 1D presaturation 1H |
| Acquisition Date: | Summer 2012 |
| Acquisition Parameters File: | nmr_data_acquisition_and_processing_protocol.doc |
| Instrument Parameters File: | Cold_adaptation_in_Drosophila_melanogaster.zip |
| Software Version: | TOPSPIN 2.1 |
| Operator Name: | Miki Watanabe and Arezue Boroujerdi |
| Chromatography ID: | CH000116 |
| Num Factors: | 12 |
| Num Metabolites: | 39 |
NMR:
| NMR ID: | NM000036 |
| Analysis ID: | AN000169 |
| Instrument Name: | Bruker Avance III |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 1D 1H |
| Field Frequency Lock: | -6958.1 |
| Standard Concentration: | 1 mM TMSP |
| Spectrometer Frequency: | 700 MHz |
| NMR Probe: | Room Temperature 5-mm triple resonance probe |
| NMR Solvent: | D2O |
| NMR Tube Size: | 5 mm |
| Shimming Method: | topshim (automatic shimming) |
| Pulse Sequence: | noesypr1d |
| Water Suppression: | presaturation |
| Pulse Width: | 9.4-9.6 ?s |
| Power Level: | -1.00 dB |
| Receiver Gain: | 200 |
| Offset Frequency: | 3294.98 Hz |
| Presaturation Power Level: | 53.64 dB |
| Chemical Shift Ref Cpd: | TMSP |
| Temperature: | 298 K |
| Number Of Scans: | 120 |
| Dummy Scans: | 4 |
| Acquisition Time: | 2.9 s |
| Relaxation Delay: | 4 s |
| Spectral Width: | 16.0872 ppm |
| Num Data Points Acquired: | 65536 |
| Real Data Points: | 32768 |
| Line Broadening: | 0.3 Hz |
| Zero Filling: | 2x |
| Apodization: | exponential multiply (em) |
| Baseline Correction Method: | abs (automatic baseline correction with integration (1D)) |
| Chemical Shift Ref Std: | 0.00 ppm |
| NMR ID: | NM000037 |
| Analysis ID: | AN000170 |
| Instrument Name: | Bruker Avance III |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 2D 1H-13C HSQC |
| Field Frequency Lock: | -6958.1 |
| Standard Concentration: | 1 mM TMSP |
| Spectrometer Frequency: | 700 MHz |
| NMR Probe: | Room Temperature 5-mm triple resonance probe |
| NMR Solvent: | D2O |
| NMR Tube Size: | 5 mm |
| Shimming Method: | topshim (automatic shimming) |
| Pulse Sequence: | HSQCEDETGPSISP2.2 |
| Pulse Width: | 9.4-9.6 ?s |
| Power Level: | f1 -1.00 dB, f2 -4.13 dB |
| Receiver Gain: | 2050 |
| Offset Frequency: | f2 3294.95 Hz, f1 14081.41 Hz |
| Chemical Shift Ref Cpd: | TMSP |
| Temperature: | 298 K |
| Number Of Scans: | 256 |
| Dummy Scans: | 256 |
| Acquisition Time: | f2 0.13317 s, f1 0.0083333 s |
| Relaxation Delay: | 1.5 s |
| Spectral Width: | f2 10.9888, f1 180.00 |
| Num Data Points Acquired: | f2 2048, f1 528 |
| Real Data Points: | f2 4096, f1 2048 |
| Line Broadening: | f2 1.0 Hz, f1 0.3 Hz |
| Zero Filling: | f2 1x, f1 4x |
| Apodization: | f2 QSINE, f1 QSINE |
| Chemical Shift Ref Std: | 0.00, 0.00 ppm |
