title: RNA-seq of mus musculus
design: For total RNA-seq, 1ml Trizol(Invitogen) were added to cells and RNA were extracted according to manufacturer's protocol. For subcelluar RNA-seq, cells were treated with buffer A (HEPES-KOH 50 mM pH 7.5, 10 mM KCl, 350 mM sucrose, 1 mM EDTA, 1 mM DTT, 0.1% Triton X-100) for 5 min on ice and then homogenized with a T 10 basic ULTRA-TURRAX homogenizer at 4th gear for 1min. The nuclei were harvested by brief centrifugation (2000g, 5 min), while the supernatant was collected as the cytoplasmic fraction. Nuclei pelltet were resuspended with same volume buffer A. Then 1ml Trizol were added to both nuclear and cytoplasmic fraction and subjected for RNA extracion. All RNA were subjected to polyA selection (Ambion, 61006) before library preparation.