RNA-Seq of Hsp70 knockdown Artemia cyst
Total RNA of 250 individuals were extracted by TriSure Reagent (Bioline) following manufacturer's procedures. The mRNA was isolated by using a bead-based Next Poly (A) mRNA magnetic isolation module (NEB, UK). Then the transcriptome library was prepared by using Ultra II RNA library preparation kit (NEB, UK). RNA was fragmented at high heat in the presence of divalent metal ions followed by cDNA synthesis and second strand synthesis. The double stranded DNA was end-repaired and ligated with Illumina adaptor followed by index PCR to incorporate sample-specific 8-bp dual barcode.  The library was quantified using Denovix High Sensitivity Kit (Denovix) and sequenced on Illumina NovaSeq 6000 platform to produce 150 bp paired-end reads