H3K27ac ChIP-Seq of HMEC-hTert breast epithelial  cells treated with Bex-Carv
Cells were cross-linked in 1% methanol-free ultrapure formaldehyde for 10 min at RT. Formaldehyde was quenched by glycine to 0.125M final concentration. The fixed cells were lysed and cell nuclei pellet was collected. Chromatin was sheared by Bioruptor Plus, diagenode sonicator. The sheared chromatin was cleared through the incubation with magnatic beads coated with nonspecific IgG antibody. Then the sheared and cleared chromatin was incubated overnight at 4C with a mixture of protein G and protein A (1:1) beads coated with antibody against H3K27ac (Cell Signaling, 8173s, Lot# 6). The protein-DNA complexes were eluted and de-crosslinked. DNA samples were purified by the addition of RNase and proteinase K. The last purification step was performed using Qiagen MinElute PCR purification kit (REF 28004).  DNA concentration was measured by the Qubit dsDNA HS assay. ChIP-seq libraries were prepared using 10 ng of DNA and Illuminas TruSeq ChIP Sample Preparation. Sequencing was performed using the Illumina's NextSeq 500 instrumental mode.