GSM7029414: 22; Homo sapiens; RNA-Seq
RNA isolation was performed with E.Z.N.A. Total RNA Kit (Omega Bio-Tek, USA) and RNeasy Plus Mini Kit (Qiagen, Germany). The cells were lysed in Lysis Buffer with beta-mercaptoethanol, and centrifuged on columns with wash buffers. RNA was eluted with RNAase-free DEPC-water. The concentration of RNA was measured on Qubit 3.0, the quality of RNA was determined by capillary electrophoresis on TapeStation (AgilentTechnologies, USA) and using R6K ScreenTape (Agilent Technologies, USA #5067-5367). RIN was 6,7-9,8. RNA libraries were prepared with NEXT flex Rapid Directional qRNA-SeqKit using indexed barcodes NEXTflex-qRNA-8nt-Barcodes (NOVA-5198-02, Bioo Scientific, PerkinElmer Applied Genomics, USA) according manufacture`s protocols. Ribosomal RNA depletion was performed with NEBNext® rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7400, New England Biolabs Inc., USA).