GSM9873918: Non_EP_5; Homo sapiens; RNA-Seq
Total RNA was extracted from frozen nasal tissue samples. Nasal polyp tissues from patients with eosinophilic CRSwNP (eCRSwNP) and non-eosinophilic CRSwNP (neCRSwNP), and inferior turbinate mucosa tissues from healthy controls were immediately rinsed with sterile saline, stored in RNase-free tubes, rapidly frozen in liquid nitrogen, and stored at −80°C until RNA extraction. The concentration and purity of total RNA were assessed using a NanoDrop-2000 spectrophotometer (Thermo Fisher Scientific, USA). RNA integrity was evaluated using an Agilent 2100 Bioanalyzer (Agilent Technologies, USA), and qualified RNA samples were used for subsequent library construction. RNA sequencing libraries were constructed using total RNA. Polyadenylated mRNA was enriched from total RNA using Oligo(dT) magnetic beads. The enriched mRNA was fragmented into approximately 300 bp fragments using ion fragmentation. First-strand and second-strand cDNA were synthesized, followed by cDNA amplification and fragment size selection. Sequencing libraries with approximately 450 bp insert sizes were obtained after library preparation and quality assessment. Indexed libraries were pooled and sequenced on an Illumina NovaSeq 6000 platform using paired-end 150 bp sequencing.