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MB Sample ID: SA206548
| Local Sample ID: | wt2d-4 |
| Subject ID: | SU002240 |
| Subject Type: | Fish |
| Subject Species: | Epinephelus coioides |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002240 |
| Subject Type: | Fish |
| Subject Species: | Epinephelus coioides |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| wt2d-4 | SA206548 | FL025290 | wild-type strain infection group | Treatment |
Collection:
| Collection ID: | CO002233 |
| Collection Summary: | The spleens of E. coioides were collected at 6, 12, 24, 48, 72 and 96 hours post injection (hpi). After phenotypic observation of spleen tissue, grouper spleen samples were collected and accurately weighed (~15 mg) and stored in a -80 °C freezer. |
| Collection Protocol Comments: | E.coioides.pdf |
| Sample Type: | Spleen |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002252 |
| Treatment Summary: | E. coioides of the infection group were intrapleurally injected with 105 colony-forming units per fish (cfu/fish) of P. plecoglossicida (wild-type strain or clpV-RNAi strain), while control E. coioides received an equivalent volume of phosphate-buffered saline (PBS). |
Sample Preparation:
| Sampleprep ID: | SP002246 |
| Sampleprep Summary: | The deep-frozen spleen tissue in each replicate was accurately weighed (about 15 mg) and then transferred to an Eppendorf tube. Each sample was first spiked with 300 μL of methanol containing internal standards and then homogenized at 65 Hz for 2.5 min. The homogenate was mixed with 1 mL of MTBE, and thoroughly vortexed for 30 min. Then, 300 μL of Milli-Q water was added to the tube, followed by vortexing for 1 min and centrifugation for 10 min to form a two-phase system (12,000 rpm, 4 ℃). Each specimen was prepared by mixing 250 μL of the top layer plus 150 μL of the bottom layer accurately and then vacuum-dried in a Speedvac concentrator (Thermo Scientific, USA). |
Combined analysis:
| Analysis ID | AN003528 | AN003529 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Waters Acquity | Waters Acquity |
| Column | Waters Acquity BEH C8 (100 x 2.1mm,1.7um) | Waters Acquity BEH C8 (100 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF-X Orbitrap | Thermo Q Exactive HF-X Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Peak area | Peak area |
Chromatography:
| Chromatography ID: | CH002606 |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters Acquity BEH C8 (100 x 2.1mm,1.7um) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003286 |
| Analysis ID: | AN003528 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | POSITIVE |
| MS ID: | MS003287 |
| Analysis ID: | AN003529 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | NEGATIVE |
