RNA-Seq of Oryctes
The total RNA was extracted from gut tissue using a Qiazol lysis reagent according to the manufacturers instructions (QIAGEN; Cat No.: 79306). The RNA samples were treated with DNase I for 1hr at 37C and then their concentrations were measured using a spectrophotometer and integrity was ensured through analysis of RNA on a 1% (w/v) agarose gel. After checking the RNA quality, total RNA samples from five different geographical regions, which passed the desired quality were pooled. A sample was submitted to Novogene Genomics Singapore Pte Ltd for RNA sequencing (RNA-Seq). To sequence the total RNA, the eukaryote ribosomal RNA was removed using a VAHTS Total RNA-seq (HMR) Library Prep Kit for Illumina.