PacBio sequencing of Mongolotettix japonicus
To construct the library for PacBio sequencing, qualified RNA extracted from each individual were mixed in equal amounts and the mixed RNA was reverse transcribed into full-length cDNA using SMARTer PCR cDNA Synthesis Kit (Clontech, CA, USA). The KAPA HiFi PCR Kits was used to amplify the full-length cDNA by PCR and BluePippin Size Selection system (Sage science, USA) was used to select the PCR product. The cDNA products with the length of 16 kb were finally retained. After repaired the end of full-length cDNA and connected the SMRT dumbbell-type connector, the cDNA products were then subjected to construction of SMRTbell Template libraries using SMRTBell Template Prep Kit.