title: GSM5230850: RNA_Ring1b_WT_pESC; Mus musculus; ssRNA-seq
library_construction_protocol: pESCs were cultured in DMEM (Kohjin-bio) with 20% fetal bovine serum (Sigma), MEM nonessential amino acids, L-glutamine, 2-mercaptoethanol, and LIF on mitomycin C-treated (Sigma) primary MEF feeder layers. For cKO, 4OHT was added to the medium to a final concentration of 800 nM. Total RNA was prepared from cultured cells using the Allprep DNA/RNA mini kit (Qiagen) following vendor's instructions. Each RNA-seq library was prepared from 1 μg of total RNA, using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB), following the supplier's instructions. Libraries were quantified with the KAPA Library Quantification Kit (Illumina). Samples were multiplexed and adjusted to 2 nM concentration, followed by Illumina HiSeq2000 sequencing. Reads were generated by 50 bp single end sequencing.