RNA-seq of Coptotermes gestroi: Termite fed with pretreated phosphoric acid sugarcane bagasse (PASCB_3)
Specimens of C. gestroi were maintained at 25 C ( 2) with 10% of humidity and fed on cardboard in appropriated boxes. One hundred workers and 10 soldiers of C. gestroi were placed in the nest chamber (with sterile sand and 10% of humidity) connected to a food chamber. As food, three different lignocellulosic materials were used: Raw Sugarcane Bagasse (SCB), Pre-treated Phosphoric Acid Sugarcane Bagasse (PASCB) and Delignified Sugarcane Bagasse (DELSCB). One gram of substrate was placed on each chamber and five replicates were established for each diet, and all experiments were maintained at 25 1C for 21 days. Afterwards, 100 workers were collected from the food chamber, frozen with liquid nitrogen and kept at -80 C until RNA extractions. Total RNA (10 g) was extracted from 50 workers of each biological replicate from the feeding experiment using Trizol reagent protocol (Thermo). The total RNA was purified using the RNeasy Plant Mini Kit (Qiagen) following manufacturers instructions. Good quality RNAs (RIN > 8.0) were submitted to rRNA depletion using the RiboZero rRNA Removal Kit under manufacturer instruction with a slight modification: at depletion step, we used a blend of rRNA removal solution from RiboZero Gold and RiboZero Bacteria kits (1:1) aiming to deplete both eukaryotic and prokaryotic rRNAs. Finally, the depleted RNAs were purified using Ampure XP beads. A total of 50 ng of depleted RNA for each replicate were used for library preparation using Stranded TruSeq RNA Sample Preparation kit, following the manufacturers instructions. For each library, paired-end sequences of 100 bp were generated in a single lane using HiSeq 2500 (Illumina).