PacBio seq of Anguilla japonica: testis
1.Oligo (dT) enrichment of mRNA containing polyA;2.Reverse transcribe mRNA into cDNA using SMARTer PCR cDNA Synthesis Kit;3.PCR amplification is used to enrich and synthesize cDNA, and the optimal conditions for PCR are determined through cyclic optimization;4.Using magnetic beads to screen fragments for large-scale PCR to obtain sufficient total cDNA content;5.Construct a full-length transcriptome library by performing damage repair, end repair, and connecting SMRT dumbbell shaped adapters using full-length cDNA;6.Nucleic acid exonuclease digestion removes the unconnected sequence at both ends of cDNA;7.Finally, by combining primers and binding DNA polymerase, a complete SMRT bell library is formed.