title: RNA-seq of Tribolium castaneum: late larvae
design: Total RNA was isolated from six larvae of ds-dy and IB groups 5 days post injection using RNAiso Plus (TAKARA, Tokyo, Japan) following the manufacturers protocol. Total RNA quantity and purity were assessed using Nanodrop 2000c and Agilent 2100 Bioanalyzer. Oligo (dT) magnetic beads were used to select Poly (A) mRNAs. After purification, the target RNA was fragmented and reverse-transcribed into double-stranded cDNA. Two specific primers were used to amplify the ligation product. The PCR product was denatured by heat, and the single strand DNA was cyclized by splint oligo and DNA ligase. Prepared libraries were then sequenced using the BGISEQ-500.