title: Illumina HiSeq 1500 paired end sequencing; Two distinct modes of DNMT1 recruitment ensure the stable maintenance DNA methylation
design: Two distinct modes of DNMT1 recruitment ensure the stable maintenance DNA methylation
library_construction_protocol: cell culture samples were collected by trypsinisation Mouse ESCs were maintained on 0.2% gelatin-coated dishes in Dulbecco's modified Eagle's medium (Sigma) supplemented with 16% fetal bovine serum (FBS, Sigma), 0.1 mM beta-mercaptoethanol (Invitrogen), 2 mM L-glutamine (Sigma), 1× MEM Non-essential amino acids (Sigma), 100 U/mL penicillin, 100 ug/mL streptomycin (Sigma), recombinant LIF (ESGRO, Millipore), and 2i (1 uM PD032591 and 3 uM CHIR99021 (Axon Medchem, Netherlands)). Column purification For RRBS, genomic DNA was isolated using a QIAamp DNA Mini Kit (QIAGEN), after an overnight lysis and proteinase K treatment. Preparation of the RRBS library was carried out as described previously (Boyle, P. et al. Gel-free multiplexed reduced representation bisulfite sequencing for large-scale DNA methylation profiling. Genome Biol. 13, R92 (2012).), with the following modifications: bisulfite treatment was performed using an EZ DNA Methylation-Gold™ Kit (Zymo Research Corporation) according to the manufacturer's protocol except that libraries were eluted in 2 x 20 µL M-elution buffer.
attributes.tag: Experimental Factor: genotype
attributes.value: PAF15KRKR