GSM9385747: ChIP_S7KO3; Mus musculus; ChIP-Seq
For anti-H3K36ac ChIP-Seq, 50,000 Spg per sample were fixed with 1% methanol-free PFA for 10'. The reaction was quenched before washing, nuclei purification and chromatin shearing in a Covaris M220 sonicator. Samples were ple-cleared and clarified by centrifugation. SNAP-ChIP K-AcylStat Panel (1/20, Epicypher) was added as a spike-in before immunoprecipitation overnight. Magnetic beads were previously blocked and conjugated with anti-H3K36ac antibody. ChIP samples were washed, crosslink reversed and ChIP / Input DNA was purified and stored at -20ºC until library preparation. Libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit for Illumina with multiplexed NEBNext Oligos (New England Biolabs). Purified Input and ChIP DNA samples were 3'-end repaired. Adaptors were ligated before PCR amplification for 13 cycles and libraries were purified and pooled for sequencing.