GSM8075759: NC-2; Homo sapiens; RNA-Seq
Total RNA was extracted from RPS27 siRNA group and RPS27 siRNA NC group using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). Following RNA extraction, the DNA was digested with DNase I. RNA quality was determined by measuring the A260/A280 ratio using a NanodropTM One/OneC spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Electrophoresis (1.5% agarose gel) was performed to confirm the RNA integrity. Qualified RNAs were quantified using the Qubit™ RNA Assay Kit (Life Technologies, Carlsbad, CA, USA). An RNA-seq library was prepared from 2 µg of total RNA using a KCTM Stranded mRNA Library Prep Kit for Illumina (cat. no. DR08402, Wuhan Seq health Co., Ltd., Wuhan, China). Polymerase chain reaction (PCR) products corresponding to 200–500 bp were enriched. For high-throughput sequencing, the libraries were prepared following the manufacturer's instructions. A NovaSeq 6000 sequencer (Illumina, San Diego, CA, USA) with a PE150 model was used for quantification and final sequencing.