GSM9893906: ARPE-19 cells, PM2.5 200 micrograms/mL, biological replicate 2; Homo sapiens; RNA-Seq
Total RNA was extracted, purified, and quality assessed by Genomics BioSci & Tech Co., Ltd. One microgram of purified total RNA from each sample was used for library preparation. The purified RNA was used for the preparation of the sequencing library by TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego, CA, USA) following the manufacturer's recommendations. Strand-specific mRNA sequencing libraries were prepared from 1 μg of purified total RNA using the TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego, CA, USA) according to the manufacturer's instructions. Briefly, polyadenylated mRNA was enriched from total RNA using oligo(dT)-coupled magnetic beads and fragmented at an elevated temperature. First-strand cDNA was synthesized using random primers and reverse transcriptase, followed by second-strand cDNA synthesis. The resulting double-stranded cDNA was purified, end-repaired, 3′-adenylated, and ligated to sequencing adapters. Adapter-ligated fragments were enriched by PCR and purified using the AMPure XP system (Beckman Coulter, Beverly, MA, USA). Library fragment size and quality were assessed using the Qsep400 System (BiOptic Inc., Taiwan), and library concentration was measured using a Qubit 2.0 Fluorometer (Thermo Fisher Scientific, Waltham, MA, USA). Qualified libraries were sequenced on an Illumina NovaSeq 6000 platform using 150-bp paired-end sequencing by Genomics, BioSci & Tech Co. (New Taipei City, Taiwan).