GSM5218728: DND41_GSI_Resistance_H3K27ac_ChIPseq_rep2; Homo sapiens; ChIP-Seq
We performed H3K27ac ChIP-seq as previously described (Petrovic et al. 2019). Briefly, we sonicated and cleared chromatin samples prepared from 107 fixed cells with recombinant protein G–conjugated Agarose beads (Invitrogen, cat# 15920-010) and subsequently immunoprecipitated these cells with antibodies recognizing H3K27ac (Active Motif, cat# 39133). We captured Antibody-chromatin complexes with recombinant protein G–conjugated Agarose beads, washed them with Low Salt Wash Buffer, High Salt Wash Buffer, LiCl Wash Buffer and TE buffer with 50mM NaCl and eluted them. After reversal of cross-linking, we performed RNase and Proteinase K (Invitrogen, cat# 25530-049) treatment and purified DNA with QIAquick PCR Purification Kit (Qiagen, cat# 28106). We then prepared libraries using the NEBNext Ultra II DNA library Prep Kit for Illumina (NEB, cat# E7645S). We validated indexed libraries for quality and size distribution using a TapeStation 2200  (Agilent). We performed paired-end sequencing (38 bp+38 bp) on a NextSeq 550.