heterozygous second instar day 3_Replicate-2
Total RNA was extracted using TrizolTM reagent (Thermo Fisher Scientific) and homogenized using a benchtop homogenizer bead and high impact zirconium beads. Library construction and high-throughput sequencing services were hired at Novogene Inc. (Sacramento, CA, USA). The libraries were sequenced using Illumina NovaSeq 6000, obtaining paired-end reads with 150 bp length and  ~50M reads per sample.