title: GSM1679862: J-20 EGC; Mus musculus; Bisulfite-Seq
library_construction_protocol: generation protocol: Genital ridges containing primordial germ cells (PGCs) were retrieved from E11.5/12.5 mouse embryos. PGCs were collected from the genital ridges by trypsinization and centrifugation. Subsequently, isolated PGCs were cultured on stem cell factor (SCF)-secreting MEF feeder containing 2i-LIF medium (Ying et al., 2008) supplemeted with 60 ng/ml exogenous SCF and 10 ng/ml bFGF.  The cells were cultured for 3 days without changing medium. After that, the cells were cultured for additional 7-11 days and fed every other day 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: GSM1679862