title: GSM1679873: J-31 ESC; Mus musculus; Bisulfite-Seq
library_construction_protocol: generation protocol: E3.5 blastocysts were retrieved from the uterine horn by flushing and collected under a dissection microscope. A whole blastocyst was plated onto MEF feeder (Globalstem) containing 2i-LIF medium (Ying et al., 2008).  The cells were cultured for 4 days without changing medium. After that, the cells were cultured for additional 3-4 days and fed every two days with fresh 2i-LIF medium until picking. Outgrowth from the culture was picked, trypsinized, and re-plated onto MEF feeders containing KO DMEM (Invitrogen) supplemented with 15% FBS (Hyclone), 1mM L- glutamine, 100 uM MEM non-essential amino acids, and 0.1 mM beta-mercaptoethanol, and LIF. Genomic DNA was collected using phenol/chloroform extraction mRRBS libraries were prepared as previously described (Boyle et al. 2012). Briefly, genomic sequences were digested using the MspI restriction enzyme, which cuts at the sequence CCGG. Digested fragments were subjected to bisulfite treatment which converts unmethylated cytosine residues to uracil
attributes.tag: GEO Accession
attributes.value: GSM1679873