Pupa
Four life stages of coconut rhinoceros beetle, Oryctes rhinoceros L., viz., early instar larva, late instar larva, pupa and adult, were sampled and total RNA was extracted from the whole body using Direct-zol RNA MiniPrep kit (Zymo, Freiburg, Germany), using manufacturers protocol. The quality and purity of the extracted RNA were assessed using an Agilent Technologies 2100 Bioanalyzer with the Agilent RNA chip and possessed RIN number (RNA Integrity Number) value > 8.0. Sequencing libraries from the resulting total RNAs were prepared (TruSeq mRNA-Seq kit and protocol from Illumina, Inc., San Diego, USA), followed by library generation (TruSeq RNA Sample Preparation Kit v2; Illumina Inc., San Diego, USA). All the four samples were sequenced using an Illumina HiSeq 2500 in rapid run mode (paired-end 100-bp reads).