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MB Sample ID: SA194793
| Local Sample ID: | SS-020 |
| Subject ID: | SU002155 |
| Subject Type: | Insect |
| Subject Species: | Drosophila melanogaster |
| Taxonomy ID: | 7227 |
| Age Or Age Range: | 3rd instar larvae |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002155 |
| Subject Type: | Insect |
| Subject Species: | Drosophila melanogaster |
| Taxonomy ID: | 7227 |
| Age Or Age Range: | 3rd instar larvae |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| SS-020 | SA194793 | FL023908 | 2/2;daGAL4/daGAL4 | Genotype of fly lines |
| SS-020 | SA194793 | FL023908 | C | Genotype Shorthand |
| SS-020 | SA194793 | FL023908 | Low Nutrient (LN) | Diet |
| SS-020 | SA194793 | FL023908 | LN | Condition |
Collection:
| Collection ID: | CO002148 |
| Collection Summary: | Ten randomly selected, wandering L3 larvae (100-115 hrs post-egg laying in SD diet; 125-140 hrs in LN diet) per biological replica (5 in total) were collected, briefly rinsed in ultrapure water, and flash-frozen in liquid N2. |
| Sample Type: | Larvae |
Treatment:
| Treatment ID: | TR002167 |
| Treatment Summary: | The fly lines were maintained on a standard (SD) or a low-nutrient (LN) diet, with a 12/12h of light/dark photoperiod cycle at 25 °C. The eggs were allowed to develop until reaching the wandering L3 larval stage for collection. The LN diet consisted of 3.5% yeast extract, 1% agar, and antimicrobials (0.1% nipagin and 0.5% propionic acid). |
Sample Preparation:
| Sampleprep ID: | SP002161 |
| Sampleprep Summary: | : Frozen larvae were homogenized in 1 ml cold (-80 °C) 80% methanol on dry ice using a probe homogenizer. Samples were clarified by centrifugation for 5 min at 16,000 x gmax at 4 °C and the supernatants were transferred to new Eppendorf tubes. The remaining pellets were resuspended in RIPA lysis buffer and total protein content was determined by the BCA method. A volume of the supernatant equivalent to 5 µg of protein was transferred to a glass vial, dried using an EZ-2 Elite evaporator (SP Scientific), and stored at -80 °C until analysis by LC-MS. |
Combined analysis:
| Analysis ID | AN003378 | AN003379 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | UltiMate 3000RSLC HPLC (Thermo Scientific) | UltiMate 3000RSLC HPLC (Thermo Scientific) |
| Column | Phenomenex Luna NH2 (150 x 2mm) | Phenomenex Luna NH2 (150 x 2mm) |
| MS Type | ESI | ESI |
| MS instrument type | Ion trap | Ion trap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | intensity (area under the curve) | intensity (area under the curve) |
Chromatography:
| Chromatography ID: | CH002497 |
| Chromatography Summary: | Separation was achieved using a linear gradient from 15% to 90% A over 18 min. This was followed by an isocratic step at 90% A for 9 min and re-equilibration to the initial 15% A for 7 min. |
| Instrument Name: | UltiMate 3000RSLC HPLC (Thermo Scientific) |
| Column Name: | Phenomenex Luna NH2 (150 x 2mm) |
| Column Temperature: | 27 C |
| Flow Rate: | 200 ul/min |
| Solvent A: | 100% water; 5 mM ammonium acetate, pH 9.9 |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003145 |
| Analysis ID: | AN003378 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Ion trap |
| MS Type: | ESI |
| MS Comments: | MS1 scan range 65-975 m/z. MS/MS with inclusion list; target others when idle. |
| Ion Mode: | POSITIVE |
| MS ID: | MS003146 |
| Analysis ID: | AN003379 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Ion trap |
| MS Type: | ESI |
| MS Comments: | MS1 scan range 65-975 m/z. MS/MS with inclusion list; target others when idle. |
| Ion Mode: | NEGATIVE |
