GSM6266272: Ngn2 Rep2.Input; Mus musculus; ChIP-Seq
Cells were collected 24h after induction, washed with PBS and fixed with 1% freshly prepared formaldehyde FA solution in PBS for 7 min at room temperature. Fixation was then quenched and cells were lysed. Chromatin was sheared using Covaris E220 High  Performance Focused Ultrasonicator. IP was performed using FLAG M2 antibody (Sigma, F1804) overnight at 4C. Samples were washed, reverse criss-linked overnight, RNase treated and NGS libraries were prepared from the isolated DNA. ChIP libraries were prepared using NEBNext Ultra-II kit (New England Biolabs) following manufacturer's protocol. Quality of the libraries was assessed with Fragment Analyzer™