Poly A RNA-Seq of Leptinotarsa decemlineata: adult whole spinosad treated insect
Samples with RNA integration number above 8 were selected for library preparation. Poly(A) enrichment was performed from 20 g of RNA to enrich for mRNA using Dynabeads mRNA Direct Micro Purification Kit. An amount of 100 ng of poly(A)-enriched RNA was fragmented with RNase III and purified using magnetic bead purification module RNA-Seq v2 kit (Life Technologies). Size distribution of the fragmented RNA was assessed on Agilent TapeStation using High Sensitivity RNA ScreenTape. A total of 50 ng of fragmented poly(A)-enriched RNA was used to prepare whole transcriptome library (RNA-Seq v2, Life Technologies). Yield and size distribution of library were analyzed on Agilent TapeStation using D1000 ScreenTape. Barcoded libraries were equally pooled and amplified onto Ion Sphere Particles (ISPs) supplied by Ion PI Hi-Q OT2 kit (Life Technologies). ISPs enriched with template library were loaded onto Ion PI chip v3 and next-generation sequencing (NGS) was performed on Ion Proton System (Thermo Fisher Scientific).