RNA-Seq of Chilo infuscatellus
Total RNA was extracted from pooled samples of 1st to 6th instar larvae using TRIzol reagent (Invitrogen, Carlsbad, CA), following the manufacturer's protocol. Small RNA sequencing was outsourced to Hangzhou Lianchuan Biotechnology Co., Ltd. Briefly, small RNA libraries were prepared using TruSeq Small RNA Sample Prep Kits, with 35 g of total RNA separated on a 15% TBE urea gel. Small RNA fragments (18-28 nt) were purified and ligated to 3' and 5' adaptors using T4 RNA ligase (Ambion). The ligated products were reverse transcribed using SuperScriptII (Invitrogen) and amplified by PCR. cDNA products were purified by gel extraction and sequenced on the Illumina HiSeq2000 platform. Post-sequencing, data were quality controlled and filtered for further analysis.