RNAseq of midgut of day 10 Sitophilus oryzae females, replicate 2
1. Biological material Sitophilus oryzae weevils are reared on wheat grains at 27.5 C and at 70% relative humidity. The Bouriz strain was because it is free of any facultative symbionts, including Wolbachia, and it harbors only S. pierantonius. For each replicate, 10 females of Day 10 were dissected to extract midgut and ovaries. ,then stored at -80 C until RNA preparation. 2. RNA isolation and libraries construction for RNAseq Total RNA was extracted with the Ambion RNAqueous-Micro kit following the manufacturer instructions. RNA was incubated with 1 U/ g of RQ1 RNase-free DNase for 30 min, at 37C then purified with RNeasy micro kit (QIAGEN) clean-up protocol. Sequencing was performed on an Illumina NextSeq500 instrument following the manufacturer's protocol and generate single-end data.