title: Second generation sequencing of Sanghuangporus sanghuang transcriptome
design: An NEBNext UltraRNA Library Prep Kit (NEB) was used to extract the total RNA from S. sanghuang according to the manufacturer's instructions. The purity, concentration, and integrity of the RNA samples were tested using Nanodrop GX to guarantee the use of qualified specimens for transcriptome sequencing. The mRNA was affluent in oligonucleotide(dT) linked magnetic beads of mRNA Capture Beads (Novozymes). Then, the mRNA was randomly divided into short segments using a fragment buffer. The template was the cleaved RNA fragment. The first strand of cDNA was synthesized by reverse transcription. Then, the second strand of cDNA was formed by adding buffer, dNTPs, RNase H, and DNA polymerase. The magnetic beads purified the double-stranded cDNA. The double-stranded cDNA was end-to-end repaired. Poly-A tails were added, ligated to the sequencing connector, and composed with DNA clean beads (Novozymes) to select cDNAs of approximately 205 bp in length. PCR amplification was performed to enrich the cDNA template. Finally, the library was sequenced.