Return to study ST000100 main page
MB Sample ID: SA005544
| Local Sample ID: | 4PC-95 |
| Subject ID: | SU000119 |
| Subject Type: | Invertebrate |
| Subject Species: | Caenorhabditis elegans |
| Taxonomy ID: | 6239 |
| Genotype Strain: | N2 |
| Age Or Age Range: | Young adult |
| Gender: | Hermaphrodite |
| Species Group: | Invertebrate |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000119 |
| Subject Type: | Invertebrate |
| Subject Species: | Caenorhabditis elegans |
| Taxonomy ID: | 6239 |
| Genotype Strain: | N2 |
| Age Or Age Range: | Young adult |
| Gender: | Hermaphrodite |
| Species Group: | Invertebrate |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 4PC-95 | SA005544 | FL001381 | 0.95 | Labelling |
| 4PC-95 | SA005544 | FL001381 | heat-shock | Experimentalgroup |
Collection:
| Collection ID: | CO000102 |
| Collection Summary: | Each flask was separated into supernatant (exometabolome) and worm pellet (endometabolome) by centrifugation 2,000 rpm for 2 min. |
| Collection Protocol ID: | Centrifugation |
| Collection Protocol Filename: | Experimental_Methods.docx |
| Sample Type: | Worms |
| Volumeoramount Collected: | 25,000 worms in 2.5 mL |
Treatment:
| Treatment ID: | TR000120 |
| Treatment Summary: | Samples were treated with a 20 min heat shock at 33 °C followed by incubation at 22C for 100 min. Sample shook at 250 rpm at a worm density of 10,000 worms per mL./ Samples were incubated at 22C for 120 min. Sample shook at 250 rpm at a worm density of 10,000 worms per mL. |
| Treatment Protocol ID: | Test/Control |
Sample Preparation:
| Sampleprep ID: | SP000115 |
| Sampleprep Summary: | - |
| Sampleprep Protocol ID: | Endometabolome |
| Sampleprep Protocol Filename: | Experimental_Methods.docx |
| Sampleprep Protocol Comments: | The worm pellets were homogenized using a FastPrep-24 (MP Biomedicals) using Lysing Matrix E tubes for 5 cycles of 60 sec in 80% methanol and then frozen at -80 °C. |
| Processing Method: | Homogenization |
Combined analysis:
| Analysis ID | AN000165 | AN000166 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS |
| Column | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units |
Chromatography:
| Chromatography ID: | CH000114 |
| Chromatography Summary: | Samples were analyzed using a mass range of m/z 70-1000 in positive and negative ionization mode, externally calibrated, using a Thermo Scientific Q-Exactive Orbitrap mass spectrometer equipped with a Dionex UltiMate 3000 RS autosampler and pump. Three ?L of each sample were injected onto an Acquity UPLC HSS T3 (2.1x100 mm, 1.8 micron) column using a column temperature of 35 C and a flow rate of 300 L/min with solvent A (0.1% formic acid in water (Fisher LC-MS Optima)) held at 100% A for 1 min, followed by linear increase to 80% B (0.1% formic acid in acetonitrile) over 10 min, kept constant for 2 min, followed by a return to initial conditions over 0.5 min and equilibration for 3 min. |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
| Flow Gradient: | A held at 100% A for 1 min, followed by linear increase to 80% B over 10 min, kept constant for 2 min, followed by a return to initial conditions over 0.5 min and equilibration for 3 min |
| Flow Rate: | 300 L/min |
| Sample Injection: | 3 ?L |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000134 |
| Analysis ID: | AN000165 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The Q-Exactive was equipped with a Heated Electrospray Ionization (HESI) source which operated at a spray temperature of 350 ?C, a spray voltage of 3.5 kV, and sheath and auxiliary gas flow rates of 50 and 10 arbitrary units, respectively. The capillary temperature was held at 325 ?C, and the S-lens RF Level was set to 40%. The FR resolution was set to 70,000 at m/z 200. The accuracy achieved was routinely less than 1.5 ppm. |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 325 ?C |
| Ion Source Temperature: | 350 ?C |
| Ion Spray Voltage: | 3.5 kV |
| Resolution Setting: | 70000 |
| MS ID: | MS000135 |
| Analysis ID: | AN000166 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The Q-Exactive was equipped with a Heated Electrospray Ionization (HESI) source which operated at a spray temperature of 350 ?C, a spray voltage of 3.5 kV, and sheath and auxiliary gas flow rates of 50 and 10 arbitrary units, respectively. The capillary temperature was held at 325 ?C, and the S-lens RF Level was set to 40%. The FR resolution was set to 70,000 at m/z 200. The accuracy achieved was routinely less than 1.5 ppm. |
| Ion Mode: | NEGATIVE |
| Capillary Temperature: | 325 ?C |
| Ion Source Temperature: | 350 ?C |
| Ion Spray Voltage: | 3.5 kV |
| Resolution Setting: | 70000 |
