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MB Sample ID: SA194791
Local Sample ID: | SS-019 |
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-019 | SA194791 | FL023908 | 2/2;daGAL4/daGAL4 | Genotype of fly lines |
SS-019 | SA194791 | FL023908 | C | Genotype Shorthand |
SS-019 | SA194791 | FL023908 | Low Nutrient (LN) | Diet |
SS-019 | SA194791 | 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 |