GSM9235697: larvae_wt_2; Drosophila melanogaster; RNA-Seq
50 ng of total RNA was used for isolation of mRNA using oligo(dT) Magnetic beads (NEB, USA) according to the manufacturer's instructions. RNA-Seq libraries were prepared with the NEBNext® Ultra RNA Library Prep kit  (NEB, USA) per the manufacturer's instructions. Purified mRNA was used as a started material for library preparation. After first and second strands synthesis cDNA was end-repaired and terminal adenosine residues were added using the NEBNext reagents. Indexed adapters were ligated, after which the material was purified with Ampure XP beads (Beckman Coulter). PCR amplification was performed using indexed primers for 12 cycles using the Q5 Hot Start HiFi PCR Master Mix (NEB). The PCR-amplified library was purified using Ampure XP beads, quantified using fluorometry with DS-11 (DeNovix, USA) and its quality was assessed on a Bioanalyzer 2100 system (Agilent).  Diluted libraries were clustered on a pair-read flowcell and sequenced using a NovaSeq 6000 system (Illumina, USA).