Small RNA-seq of Ostrinia furnacalis egg masses and adult's gonads

Total RNAs were prepared using Trizol reagent (Invitrogen) according to the manufacturer's protocol. Small RNA was prepared from total RNA as reported previously (Shoji et al., 2017). Ten to 15 micrograms of total RNA was separated by electrophoresis in denaturing 24% polyacrylamide gels containing 4 M urea, and stained with SYBRGold (Invitrogen). Small RNA fractions containing around small RNAs of 20 to 30 nts were recovered using the ZR small-RNA PAGE Recovery Kit (ZYMO Research). Library construction of small RNA was performed by using TruSeq RNA Sample Prep Kits (Illumina) for egg samples and by using Small RNA Cloning Kit (TaKaRa) for gonad samples , according to the manufacturer's protocols. Sequencing was carried out with the Illumina HiSeq 2500 platform.