Protocols: P2AIII and attP2 (control) were crossed to uas-rpr (X). Crosses were put in collection cages containing apple juice agar plates at kept at 25oC in a 12h light/dark cycling incubator. 1st instar larvae were picked from apple agar plates and placed into vials containing standard fly at 100 larvae/15mL vial. Larvae were reared at 25oC in a 12h light/dark cycling incubator until further experimentation. 10 male larvae were collected at 3rd instar wandering stage across independent replicates per sample and snap-frozen in liquid N2. 10 male larvae were collected across independent replicates per sample and snap-frozen in liquid N2. Larvae were transferred to Lysing Matrix E homogenization tubes with 1ml of TRIzol and ruptured on high settings inf a FastPrep tissue homogenizer. The supernatant was transferred to a fresh tube and spun down for 2 minutes at max speed. 800µl of TRIzol sample was transferred to prespun phaselock tube and 200µl Chloroform added. Phases were separated by spinning at 12000 rcf, 15 min, 4°C. Upper aqueous phase was transferred to a fresh tube and mixed with 500 µl isopropanol. The resulting mix was spun at 20000 rcf, 15 min, 4°C. All supernatant was then drained and the pellet resuspended in 30 µl DNAse solution (Ambicon, final conc. 0.2U/µl) and incubate for 1h at 37°C. RNA was purified using the RNeasy Plus kit by addid 270µl RTL buffer and then isolating according the the manufacturers instructions. Libraries were generated by the Max Planck Genome Centre (Cologne). Libraries were stranded, from polyA enriched RNA.
ENA-FIRST-PUBLIC
2022-09-14
ENA-LAST-UPDATE
2022-09-14
External Id
SAMEA10535033
INSDC center alias
Max Planck Institute for Infection Biology, Cellular Microbiology
INSDC center name
Max Planck Institute for Infection Biology, Cellular Microbiology
INSDC first public
2022-09-14T20:17:34Z
INSDC last update
2022-09-14T20:17:34Z
INSDC status
public
Submitter Id
E-MTAB-11095:7 attP2 control rep3
age
96
broker name
ArrayExpress
common name
fruit fly
developmental stage
third instar larva stage
genotype
attP2 control
organism part
whole organism
phenotype
wild type
sample name
E-MTAB-11095:7 attP2 control rep3
sex
male
strain
yw