RNA-Seq of Lentinula edodes strain 1303: untreated control mycelia (replicate 2)
First, mRNA was enriched from total RNA using oligo(dT) magnetic beads. The purified mRNA was randomly fragmented into short segments using divalent cations under elevated temperature. Using the fragmented RNA as a template and random oligonucleotides as primers, double-stranded cDNA was synthesized. The purified double-stranded cDNA underwent end repair, dA-tailing, and sequencing adapter ligation. The ligation products were purified and subjected to size selection to obtain cDNA fragments of the desired length. Subsequently, PCR amplification was performed, and the PCR products were purified again to obtain the final sequencing library.