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MB Sample ID: SA005520
Local Sample ID: | 6PC-5 |
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 |
---|---|---|---|---|
6PC-5 | SA005520 | FL001379 | 0.05 | Labelling |
6PC-5 | SA005520 | FL001379 | 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 |