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MB Sample ID: SA027281
| Local Sample ID: | 20150724_YU_2_1_2 |
| Subject ID: | SU000544 |
| Subject Type: | Animal |
| Subject Species: | Megalobrama amblycephala |
| Taxonomy ID: | 75352 |
| Species Group: | Fish |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000544 |
| Subject Type: | Animal |
| Subject Species: | Megalobrama amblycephala |
| Taxonomy ID: | 75352 |
| Species Group: | Fish |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 20150724_YU_2_1_2 | SA027281 | FL006865 | 2 years old sexually mature without hormone treatment | Phenotype |
Collection:
| Collection ID: | CO000538 |
| Collection Summary: | About 2 ml of blood was collected via caudal vein by syringe for each female M.amblycephala and then directly put into K3 EDTA centrifuge tube. |
| Collection Protocol Filename: | Zhou999_20161213_161728_PR_CO_collection_files.docx |
| Sample Type: | Plasma |
Treatment:
| Treatment ID: | TR000558 |
| Treatment Summary: | The blood was centrifuged at 4000 rpm for 5 min at 4°C as soon as it was extracted from fish and about 1 ml supernatant was pipetted into another tube.The extracted plasma samples were then frozen in liquid nitrogen immediately and stored in the -80°C. |
Sample Preparation:
| Sampleprep ID: | SP000551 |
| Sampleprep Summary: | To initiate the experiment, each plasma sample was mixed with the same volume and labeled as Quality control (QC) sample. Plasma sample (100 µl, including QC sample) was added into new Eppendorf tube with ice-cold methanol (300 µl), vortex mixing for 1 min, standing for 1 min, and make sure the solution volume had no obvious observable difference. Then samples were precipitated for 20 min at -20°C, and centrifuged at 14000 rpm for 20 min at 4°C to precipitate the proteins. A total of 250 µl of protein free supernatant was collected and dried with nitrogen at 37°C. The concentration of extractant was diluted depending on the mass spectrum response signals. |
| Sampleprep Protocol Filename: | Zhou999_20161213_161728_PR_SP_sampleprep.docx |
Combined analysis:
| Analysis ID | AN000796 | AN000797 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Waters 2777C | Waters 2777C |
| Column | Waters Acquity BEH C18 (150 x 2mm,1.7um) | Waters Acquity BEH C18 (150 x 2mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Waters Synapt G2 XS QTOF | Waters Synapt G2 XS QTOF |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | peak intensity | peak intensity |
Chromatography:
| Chromatography ID: | CH000572 |
| Chromatography Summary: | Chromatographic separation was performed on a ACQUITY UPLC BEH C18 analytical column (100×2.1 mm, 1.7 µm, Waters, Milford, USA) using a 2777C UPLC system (Waters). The flow rate was 0.40 mL/min and the mobile phase was composed of solvents A (95% H2O/5% acetonitrile + 0.1% formic acid) and B (95% acetonitrile/5% H2O＋0.1% formic acid).The gradient program was optimized as follows: 0–0.1 min, 100% A; 0.1–0.6 min, 100% A-50% A; 0.6–5 min, 50%–0% A; 5–8 min, 0% A; 8–10 min, 0% A-100% A. Injection volume was 10 µl. The eluent from the column was directly added in to the mass spectrometer without split. |
| Instrument Name: | Waters 2777C |
| Column Name: | Waters Acquity BEH C18 (150 x 2mm,1.7um) |
| Flow Gradient: | 100% acetonitrile |
| Flow Rate: | 0.40 mL/min |
| Solvent A: | 95% water/5% acetonitrile; 0.1% formic acid |
| Solvent B: | 95% acetonitrile/5% water; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000703 |
| Analysis ID: | AN000796 |
| Instrument Name: | Waters Synapt G2 XS QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| MS ID: | MS000704 |
| Analysis ID: | AN000797 |
| Instrument Name: | Waters Synapt G2 XS QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
