title: RNA-Seq of Panchlora nivea: adult male whole body
design: Total RNA was isolated.  mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. Fragmentation was carried out using divalent cations under elevated temperature. First strand cDNA was synthesized using random hexamer primer. Second strand cDNA synthesis was performed using DNA Polymerase I and RNase H. NEBNext Adaptor with hairpin loop structure were ligated to prepare for hybridization. cDNA fragments of preferentially 150~200 bp in length, were selected. Then PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer. PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 system