GSM9248408: Kc167, CP190 R1, CTCF R1, Pc R1 Input in mock cells, bio rep 1; Drosophila melanogaster; ChIP-Seq
2–4×107 cells were fixed by adding 1% formaldehyde directly to cells in culture medium for 10 min at RT with gentle agitation. 0.125 M glycine was used to quench formaldehyde with gentle agitation for 5 min at RT. Cells were centrifugated at 600 x g for 5 min and washed twice with ice-cold PBS. Chromatin preparation and chromatin immunoprecipitation (ChIP) were performed following earlier described methods (Bag et al., 2021), with minor modifications to the chormatin sonication steps. The modifications are as follows, cells were incubated in lysis buffer for 10 min on ice before sonication, which was completed using the Bioruptor (Diagenode) using 15 cycles of 30s on and 30s off. Libraries were prepared according to the manufacturer's protocol (NEB; Ultra II for DNA Library Prep Kit; E7645).