GSM8174705: PVR_K36me3_2; Mus musculus; ChIP-Seq
Isolated primary retinal pigment epithelium cells were lysed in lysis buffer (0.5% Tween-20, 0.1% SDS, 0.5% NP-40, and Protease Inhibitor Cocktail) for 10 min on ice. Then MNase buffer (2 mM CaCl2, 100 mM Tris-HCl pH 8.0) containing 0.02 U MNase was added and incubated at 37°C for 5 min. The reaction was terminated by adding stop buffer (55 mM EDTA, 110 mM Tris-HCl pH 8.0) and incubated with 2x RIPA buffer (280 mM NaCl, 0.1% SDS, 1% Triton X-100, 5 mM EGTA, 0.2% sodium deoxycholate, and Protease Inhibitor Cocktail) for 10 min on ice. Then the sample was centrifuged at 14500 rpm for 15 min, the supernatant was collected and added with RIPA buffer (10 mM Tris-HCl pH 8.0, 1 mM EDTA, 140 mM NaCl, 0.1% SDS, 1% Triton X-100, and 0.1% sodium deoxycholate). Immunoprecipitation was performed using 1 μg antibody. The library preparation according to the VAHTS Universal DNA Library Prep Kit for Illumina V3 Kit (ND607, Vazyme)