GSM6235400: Microglia H3K27me3 rep1; Homo sapiens; ChIP-Seq
10x10^6 million microglia cells were fixed with 1% formaldehyde/PBS  followed by quenching with 0.125 M glycine. After washing with  cold PBS, pellet was resuspended in swelling buffer, cytosol was removed and nuclei sonicated. Lysates were cleared and 2-8 microgram of chromatin were incubated with corresponding amounts of antibody overnight at 4°C. The next day, chromatin was captured with Protein A/G beads, washed and eluted with TE buffer, 1% SDS, 100 mM NaCl. 0.5 mg/ml Proteinase K were added, and samples were incubated for 2 h at 55°C and overnight at 65°C. The next day, RNA was digested. Supernatant was removed from the magnetic beads and DNA was purified with AMpure beads XP clean up according to manufacturer's instructions. ChIP libraries were prepared using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®  (NEB) and NEBNext® Multiplex Oligos for Illumina® (Index Primers Set 1) (NEB) according to manufacturer's instructions.