GSM7816469: dease4; Homo sapiens; RNA-Seq
Total RNA was isolated from PBMCs using the Magzol Reagent (Magen, China), as per manufacturer's protocol. The quantity and integrity of the RNA was assessed using the K5500 micro-spectrophotometer (Beijing Kaiao, China) and the Agilent 2200 TapeStation (Agilent Technologies, USA), respectively. Briefly, rRNAs were removed from the total RNA using the QIAseq FastSelect-rRNA HMR Kit (QIAGEN, Germany). The RNA was then treated with RNase R (Epicenter, USA) and fragmented to approximately 200 bp the purified RNA fragments were subjected to first strand and second strand cDNA synthesis, followed by adaptor ligation and enrichment with a low cycle according to manufacturer's instructions for the NEBNext® Ultra™ RNA Library Prep Kit for Illumina (NEB, USA). The library products were evaluated using the Agilent 2200 TapeStation and Qubit (Thermo Fisher Scientific, USA), and further sequenced using an Illumina (Illumina, USA), through the paired-end 150 bp approach, at Ribobio Co. Ltd. (Ribobio, China).