GSM7989034: Expression of AKT1 WT; Drosophila melanogaster; RNA-Seq
Ovaries were dissected in S2 medium (Schneider's insect medium supplemented with 10% fetal calf serum). Ovaries (30 pairs) were washed three times in calcium-free phosphate buffered saline (PBS) and incubated at 22°C for 30 min with vigorous shaking (2000rpm) in 1 ml of 0.05% trypsin and 5mg/mL collagenase in PBS. Supernatant was removed, filtered through a 30µM nylon mesh into tubes containing 1 ml of S2 medium, pelleted at 1000g for 5 min in a Microcentrifuge, and washed in 1 ml of PBS. For RNAseq experiment, the pellets were resuspend directly in TRI-reagent and store at -80°C. RNA extraction was performed using the TRI-reagent supplier's instructions (Molecular Research Center, Cincinnati, Ohio, USA). TruSeq mRNA stranded Illumina