GSM5625442: RNAseq_shCtrl_Rep3; Mus musculus; RNA-Seq
RNA samples were extracted directly from cultured cells using Trizol reagent (Invitrogen) followed by isopropanol precipitation, and sample quality control was performed with Agilent Bioanalyzer 2100. All of the samples had an RNA integrity number ranging from 9.9 to 10. For RNA-seq library preparation, 1 μg of total RNA was depleted of ribosomal RNA using the RiboMinus Eukaryote System v2 kit (Invitrogen), following manufacturer instructions. Ribo-RNA was resuspended in 17 μL of EFP buffer (Illumina), heated to 94 °C for 8 min, and used as input for the first-strand synthesis, using the TruSeq RNA Sample Prep kit, following manufacturer instructions. Poly(A) RNA-seq library was performed by using the TruSeq RNA Sample Prep kit, following the manufacturer's instructions.