full-length transcriptome of Misgurnus bipartitus: multi tissues
Total RNA was extracted from tissue samples, and the concentration, purity and integrity of the RNA were examined by instruments. Use Clonetech SMARTerTM PCR cDNA Synthesis Kit to synthesize the full-length cDNA of mRNA. A primer with Oligo dT is used for A-T base pairing with polyA as a primer for reverse synthesis of cDNA, and a primer is added to the end of the reverse synthesized full-length cDNA. The obtained full-length cDNA was amplified by PCR, and the product was purified by PB magnetic beads and quantified by Qubit 3.0. Purify the amplified full-length cDNA with PB magnetic beads, and remove some small fragments of cDNA below 1kb. End-repair the full-length cDNA and connect the SMRT dumbbell linker. Perform exonuclease digestion of the fragments that are not connected to the adapter, and then use PB magnetic beads for purification again to obtain a sequencing library.