title: GSM2405855: Input_Hippocampal_neurons_pooled; Mus musculus; MeDIP-Seq
library_construction_protocol: C57BL/6J (Janvier Labs, France) hippocampi of E18.5 embryos were dissected and collected in 15 ml Hanks’ Balanced Salt Solution (HBSS, PAA, Germany). After centrifugation (5min; 800xg) hippocampi were dissociated by adding 1ml 0.05% trypsin-ethylenediaminetetracetic acid (PAA) at 37°C for 15min. After adding 1ml of fetal bovine serum (FBS, Life Technologies) and 1ml of neurobasal (NB) medium supplemented as previously described (Vezzali R et al. 2016) samples were homogenized by repeated passages through a pipette tip. After 10min centrifugation at 1000Xg, cells were resuspended in complete NB medium containing 22µM L-glutamic acid (Sigma, Germany), counted, and seeded on poly-ornithine (0.1 mg/ml, Sigma) and laminin (1μg/ml, Sigma) coated plates. Medium change was performed every 96h by exchanging half of the volume of the medium with fresh NB medium with complements and AraC (1µg/ml for the first, and 0.5µg/ml for the second medium change, Sigma). At day in vitro (DIV) 11 cells were treated with 55mM KCl at 37ºC, 5%CO2 at different time points before harvesting. The company Active motif processed the samples, constructed the library and analyzed the data.
identifiers.namespace: GEO
identifiers.id: GSM2405855