characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-04-13T16:27:35Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-04-13T16:27:35Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112653207
characteristics.characteristic: External Id
characteristics.text: Department of Ecology & Evolution University of Luasanne
characteristics.characteristic: INSDC center name
characteristics.text: 2023-04-13T16:27:35Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-04-13T16:27:35Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14650918
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12656:MEL_LE_F_2
characteristics.characteristic: Submitter Id
characteristics.text: 2 to 10
characteristics.unit: day
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: fruit fly
characteristics.characteristic: common name
characteristics.text: Protocols: All adult samples were collected from flies aged between 2-10 days. Antenna were collected by flash freezing flies in liquid nitrogen and agitating them over a sieve connected to a collection dish. Antenna were selected from the collection dish using a pipette under a dissecting scope. Forelegs and proboscis with maxillary palps were collected from individual files using forceps and microscalpels under a dissecting scope. Third instar larva were collected from vials by floating them in 75% sucrose water and washed. Larva heads were removed under a dissecting scope using a microscalpel. For each replicate, ~10 third instar larval heads, ~25 proboscis, ~50 legs, ~5 ovipositors and ~100 antennae were collected. After sorting sexes on C02, all flies were left in food vials for at least 24h before the dissections. All adult samples were collected from flies aged between 2-10 days. Antenna were collected by flash freezing flies in liquid nitrogen and agitating them over a sieve connected to a collection dish. Antenna were selected from the collection dish using a pipette under a dissecting scope. Forelegs and proboscis with maxillary palps were collected from individual files using forceps and microscalpels under a dissecting scope. Third instar larva were collected from vials by floating them in 75% sucrose water and washed. Larva heads were removed under a dissecting scope using a microscalpel. For each replicate, ~10 third instar larval heads, ~25 proboscis, ~50 legs, ~5 ovipositors and ~100 antennae were collected. After sorting sexes on C02, all flies were left in food vials for at least 24h before the dissections. Fly strains were reared on a standard yeast/cornmeal/agar medium supplemented with Carolina 4-24 Formula and maintained in a 12:12 hr light:dark cycle at 25 degrees. Adults between 2 to 10 days old were sex-sorted on CO2 at least 24h before the dissections. Dissected tissues were homogenized in 200ul of Trizol (Invitrogen) using a Precellys24 (6800rpm, 2x30s with 10s breaks ; Bertin Technology) followed by a standard Trizol RNA extraction. The final mRNA concentration was measured using a DeNovix Ds-11 FX spectrophotometer. mRNA libraries were prepared using KAPA Stranded mRNA-seq Kit (Roche) following manufacturer’s instructions (Version 5.17). Briefly, 500ng of total RNA diluted in 50ul of RNAse-free water was first placed on supplied mRNA capture magnetic beads to allow the isolation of mature, polyadenylated, mRNA which was subsequently fragmented to a size of 100-200bp. Double-strand cDNA was then synthesized, marked by A-tailing and barcoded with 2.5ul of TruSeq RNA UD Indexes (Illumina). SPRI select beads (Beckman Coulter) were used for cleanup.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: Drosophila melanogaster
characteristics.characteristic: organism
characteristics.text: Drosophila melanogaster
characteristics.characteristic: organism
characteristics.text: foreleg
characteristics.characteristic: organism part
characteristics.text: Drosophila melanogaster
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: B54
characteristics.characteristic: strain
characteristics.text: MEL_LE_F_2
characteristics.characteristic: title