GSM8153321: Cfra_H3K4me3_rep1; Creolimax fragrantissima; ChIP-Seq
Fixed cell pellets are treated with 2 ul of 3%SDS, to achieve a final concentration of 0.5% SDS. Cells are thoroughly re-suspended and lysed on ice for 10 min. Lysates are transferred into 0.1 ml Bioruptor Microtubes (Diagenode C30010015, Liège, Belgium) and the chromatin was sheared using an NGS Bioruptor Sonicator (Diagenode) at High Intensity and cycles of 30'' ON/30'' OFF. 100 ng chromatin per species was pooled per ChIP. Pooled chromatin was incubated at 4°C for 12−14 h with rotation with 2.5 µg anti-H3K27ac (Abcam, ab4729) or  2.5 µg anti-H3K4me3 (Millipore, 07-473). Immunoprecipitated complexes were captured using a mix of Protein A (16-661, Sigma-Aldrich) and Protein G magnetic beads (16-662, Sigma-Aldrich), washed, and reverse crosslinked for 30 min at 55°C followed by an hour at 68°C. Immunoprecipitated DNA was purified using SPRI beads (A63881, Beckman Coulter). The NEBNext Ultra II DNA Library Prep Kit (New England BioLabs) was used to produce the resulting ChIP libraries according to the manufacturer's protocol.