Transcriptome Analysis of Dastarcus helophoroides (Coleoptera: Bothrideridae) Using Illumina HiSeq Sequencing
Total RNA for transcriptome analysis was isolated from whole adults of D. helophoroides using Trizol reagent (Sangon Biotech, Shanghai, China). Total RNA was treated by DNase I to remove DNA. Next, magnetic beads with Oligo (dT) were used to isolate mRNA. The mRNA was mixed with fragmentation buffer and fragmented into short fragments. Next, the cDNA was synthesized using the mRNA fragments as templates. Short fragments were purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. Next, the short fragments were connected with adapters. Suitable fragments were selected for PCR amplification as templates.Finally, the library was sequenced using the Illumina HiSeq 2000 sequencer (Beijing Genomics Institute, BGI, Shenzhen, Guangdong, China). After sequencing, raw image data were transformed by base calling into sequence data, which were called raw data or raw reads and were stored in the fastq format.