title: GSM8945963: ChipSeq_FNP_E11.5_WT_H3K27ac; Mus musculus; ChIP-Seq
library_name: GSM8945963
library_construction_protocol: Dissected tissue from E8.5 embryos (i.e. dorsal most portion of the neural plate) and from E10.5 and E11.5 embryos (i.e. FNP, Mx, Md and PA2) were micro-dissected and incubated in trypsin 0.5%/EDTA 1X at 37 °C for 10 minutes, rinsed (DMEM 1X/ FBS 10%, followed by two washes in ice-cold 1× DMEM), dissociated by pipetting, cross-linked with 1% formaldehyde in PBS 1X for 10 minutes at room temperature (RT) and quenched with glycine (final concentration 125 mM) for 5 minutes at RT. Cells were spun down (1600 rpm, 10 minutes, 4 °C) and rinsed in ice-cold PBS. CNCCs from E11.5 Wnt1::Cre;ROSARFP and Wnt1::Cre;ROSARFP;Ezh2 flox/flox (Ezh2cKO) embryos were filtered and collected by FACS. ChIP-seq experiments with anti-H3K4me2 (Millipore, Cat. #07-030), anti-H3K27me3 (Millipore, Cat. # 07-449), anti-H3K27ac (abcam, ab4729) and anti-CTCF (Cell Signaling, CST 2899) antibodies were performed as described in Minoux et al., 2017 and Kessler et al., 2023. ChIP-seq libraries were generated using bar-coded adapters (NEB, E7335) in combination with the NEBNext® Ultra™ DNA Library Prep Kit for Illumina® (NEB, E7370) according to manufacturers' instructions. The quality of the libraries and size distribution was assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies). Sequencing was performed on an Illumina HiSeq 2500 machine (50 bp read length, single-end) or an Illumina NovaSeq 6000 machine (50 bp read length, paired-end) .
identifiers.namespace: GEO
identifiers.id: GSM8945963_r1