ChIP-seq (histone modification H3K27ac) of human: PBMCs 5
Cells were homogenized and chromatin cross-linked with formaldehyde (final concentration 1%) for 10 minutes. The cross-linking reaction was stopped by adding 2 M glycine (final concentration 0.125 M), and the samples were incubated for 5 minutes. Chromatin was fragmented using a sonicator to obtain fragments 150-500 bp long. Chromatin immunoprecipitation was performed with antibodies to histone modifications H3K4me3 or H3K27ac for 6 hours at +4 C. Next, chromatin was washed from nonspecific products and DNA was extracted using phenol-chloroform. DNA libraries were prepared according to the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina protocol. Paired-end sequencing was performed on a NovaSeq 6000 platform (Illumina).