NextSeq 2000 paired end sequencing; ChIP seq for H4K5ac, JunD , and MED1 in Parental and Paclitaxel resistant L3.6pl cell line
cells were crosslinked using 1% formaldehyde for 20 minutes and nuclear pellets were sonicated in sonication buffer (150mM NaCl, 20 mM EDTA, 50 mM Tris-HCl (pH 8), 1% v/v NP-40, 0.5% v/v sodium deoxycholate, 20 mM NaF, 0.1% SDS) for 30 cycles using a Bioruptor Pico (Diagenode) and a cycle setting of 30 s on/off. Samples were precleared by 50% slurry of Sepharose 4B (GE Healthcare) and incubated with antibody overnight DNA was extracted after decrosslinking using Phenol/chloroform/isoamyl. Libraries were prepared using the MicroPlex Library Preparation Kit v2 (Diagenode) according to manufacturerâ€™s protocol.