GSM4655034: PHH_HUM183221; Homo sapiens; RNA-Seq
Total RNA was extracted from cultured cells, using a PureLink RNA Mini Kit (Thermo Fisher Scientific). A cDNA library was generated from 10 ng of total RNA, using a SuperScript VILO cDNA Synthesis Kit (Thermo Fisher Scientific). Amplification, primer digestion, and adapter ligation were performed using an Ion AmpliSeq Transcriptome Human Gene Expression Kit (Thermo Fisher Scientific). The cDNA library was purified using Agencourt AMPure XP Reagent (Beckman Coulter) and quantified using Ion Library TaqMan Quantitation Kit (Thermo Fisher Scientific), followed by dilution to 75 pM with water and pooled equally. Eight samples per pool were sequenced using an Ion 540 Chip Kit (Thermo Fisher Scientific) simultaneously, using IonS5 XL (Life technologies) and Ion Chef instrument systems (Life technologies) with Ion 540 Kit-Chef (Life technologies). All procedures were performed per the manufacturers' protocols.