title: GSM5336286: ChIPseq_ESC_Ngn2_uninduced_IgG_r3; Mus musculus; ChIP-Seq
library_construction_protocol: Three 15 cm dishes per ChIP-seq replicate were seeded (6x106) for neural generation three days before the experiment. Chromatin Immunoprecipitation (ChIP) was carried out as previously described (Lienert, F. et al. Nat. Genet., 2011) with following modifications: (1) Neurons were washed once with PBS, collected in PBS and cross-linked in 10ml PBS with 1% Formaldehyde (Sigma, 252549-25ml) for 10 min at room temperature, (2) chromatin was sonicated for ~22 cycles of 20 sec. on and 40 sec. off using a Diagenode Bioruptor Pico, (3) 75-200ug of chromatin was used per IP, (4) protein A/G magnetic Dynabeads (Thermo Fisher Scientific) were used.