RNA-Seq of Volvariella volvacea
After the sample is tested and found to be qualified, magnetic beads coated with Oligo(dT) are used to enrich the mRNA of eukaryotes. Then, a fragmentation buffer is added to randomly break the mRNA. Using the mRNA as a template, the first strand of cDNA is synthesized with six-base random primers (random hexamers), and then a buffer, dNTPs, and DNA polymerase I are added to synthesize the second strand of cDNA. Subsequently, the double-stranded cDNA is purified using AMPure XP beads. The purified double-stranded cDNA undergoes end repair, addition of an A tail, and ligation of sequencing adapters. Finally, size selection of the fragments is performed using AMPure XP beads, and then PCR enrichment is carried out to obtain the final cDNA library.