2.11. RNA‐seq library construction and data processing

Total RNA of WT and Rnf20 knockout reprogrammable cells at day 3 was harvested using Trizol (Invitrogen). RNA‐seq libraries were prepared with AMPure XP beads mRNA Purification Kit (Beckman) and TruSeq Stranded mRNA Library Prep Kit (Illumina). Sequencing was carried out with Illumina HiSeq 2000. Raw paired‐end sequencing reads were aligned to mouse GRCm38/mm10 reference genome by HISAT2 (2.1.0). 26 FPKMs (fragments per kilobase of transcript per million mapped reads) were calculated using Cufflinks (v2.1.1). The genes with FPKMs>1, fold changes >2 and FDR<0.01 were identified as the differentially expressed genes. GO analyses were performed using DAVID 27 and WebGestalt. 28 Table S2 contains the datasets of RNA‐seq.