GSM6255471: H2A.Z (Chd4 WT, Rep2); Mus musculus; ChIP-Seq
mES cells were fixed using 1% formaldehyde and lysed in ChIP lysis buffer (50 mM Tris PH 8.0, 100 mM NaCl, 5 mM EDTA PH 8.0, 0.5% SDS, 50 mM NaF, 20 mM b-glycerophosphate and protease inhibitor cocktail). Nuclei were isolated by spin down at full speed for 15 min and suspended in IP buffer (A 2:1 mix 2 of ChIP SDS lysis buffer and ChIP dilution buffer containing 100 mM Tris PH 8.0, 5 mM EDTA PH 8.0 and 5% Igepal CA630) followed by chromatin shearing to 200-500b and chromatin immunoprecipitation using indicated antibodies and protein G Sepharose beads. Precipitated chromatins were isolated by decrosslinking and purification with Qiaquick PCR purification kit and subjected to sequencing library preparation directly. DNA libraries were prepared following NEBNext Ultra II DNA Library Kit for Illumina (New England Biolabs) guidelines.