title: RNAseq of setd2KO mESC
design: RNAseq of setd2KO mESC
library_name: 21-028_4_ESC_SN_p
library_construction_protocol: Naive murine embryonic stem cells (mESC) were derived freshly (FVB or mixed 129/B6)  and were routinely cultured on gelatin-coated plates in t2i/L media: NDiff (N2B27) (Takara #y40002) supplemented with titrated 2i (0.2 uM PD0325901 and 3 uM CHIR 99021), 1,000 U/ml leukaemia inhibitory factor (LIF), 1% FBS, 1% penicillin streptomycin and maintained in humidified atmosphere at 37C and 5% CO2. Cells were passaged every 2 days by dissociation with TrypLE and medium was changed daily. RNA-sequencing of total RNA from mouse Embryonic Stem Cells.  Total RNA was isolated using the Monarch. Total RNA Miniprep Kit (NEB #T2010S), including gDNA removal step and on-column DNase I digestion.  Total RNA was quantified using Qubit II and quality assessed with high-sensitivity RNA Screen Tape (Agilent 5067-5579).  rRNA was depleted using the NEBNext rRNA Depletion Kit v2 (Human/Mouse/Rat) (cat.no. E7400X)  Stranded RNA-seq libraries were prepared with the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (cat.no. E7760L) (input: 500 ng of total RNA) following the manufacturer's instructions.