RNA-seq Analysis of Pleurotus eryngii

Total RNA was extracted using TRIzol, according to the manufacturer’s instructions (Invitrogen, Carlsbad, CA, USA). The quality of the isolated RNA was detected by a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The mRNA was purified, and the cDNA library was constructed according to the method of Illumina RNA seq libraries of Shanghai Meiji Biomedical Technology (Shanghai, China). Sequencing was performed on an Illumina Novaseq 6000. The raw sequencing data were filtered using fastp software to obtain high-quality data (clean data) for subsequent analysis [30 (link)]. The filtered data were aligned to the reference genome (reference species: Pleurotus eryngii; reference genome version: GCA_015484515.1; reference genome source: https://www.ncbi.nlm.nih.gov/genome/45890?genome_assembly_id=1500182 accessed on 20 February 2022), and the data comparison results were counted and evaluated again after quality control, mainly including sequencing coverage situation analysis. Three biological replicates were used for the RNA-seq experiments.