Global Transcriptome Profiling of the Pine Shoot Beetle, Tomicus yunnanensis (Coleoptera: Scolytinae)
Total RNA was extracted from each 20 female and male adult beetles using TRIzolH Reagent (Invitrogen) according to the manufacturer’s instructions. RNA quality and yield were assessed by 2100 Bioanalyzer (Agilent technologies) with a minimum RNA integrated number value of 8. The cDNA library was prepared using Illumina’s kit following manufacturer’s recommendations. Briefly, messenger RNA was isolated from 20 mg total RNA (pooled RNA of female and male adults) using oligo(dT) magnetic beads and fragmented into short sequences using divalent cations under elevated temperature. First and second strand cDNA were synthesized from cleaved RNA fragments. After the end repair and ligation of adaptors, the products were cleaned up with a QIAquick PCR Purification Kit to create the final cDNA library.