RNA-Seq of Agrilus zanthoxylumi: adult male abdomen
After the total RNA sample is qualified, the mRNA is enriched with magnetic beads with Oligo (dT), and a fragment buffer is added to the obtained mRNA to make the fragment into short fragments. The fragmented mRNA is used as a template and six bases are used. Random primers were used to synthesize the first strand of cDNA, and buffers, dNTPs, RNaseH, and DNA Polymerase I were added to continue to synthesize the second strand of cDNA, purified by QIAQuickPCR kit and eluted with EB buffer. The purified double-stranded cDNA is eluted and then subjected to end repair, base A, and sequencing adapter treatment, and then the target size fragment is recovered by agarose gel electrophoresis and PCR amplified to complete the entire library preparation. The constructed library was sequenced using the Illumina platform, and the sequencing strategy was PE150.