Dual RNAseq of bacteriome of 4th stage larvae Sitophilus oryzae, replicate 3
Total RNA was extracted with TRIzol Reagent (Invitrogen, ref.: 15596026) following the manufacturers instructions. Nucleic acids were then purified using the NucleoSpin RNA Clean up kit (Macherey Nagel, ref.: 740948). Genomic DNA was removed from the samples with the DNA-free DNA removal kit (Ambion, ref.: AM1906). Total RNA concentration and quality were checked using the Qubit Fluorometer (Thermo Fisher Scientific) and Tapestation 2200 (Agilent Biotechnologies). Ribo-depletion and dual RNA-seq strand-specific cDNA libraries were obtained starting from 100 ng of total RNA using the Ovation Universal RNA-seq System (NuGEN) following the manufacturers instructions. Libraries were sequenced on a Nextseq 500 sequencer (Illumina), using the NextSeq 500/550 High Output Kit (Illumina).