GSM1625984: ChIP-Seq of H3K27me3 in TPP Macrophages; Homo sapiens; ChIP-Seq
Chromatin from 0.5x10E6 macrophages was cross-linked for ChIP reactions with 1 % formaldehyde. Nuclei were lysed and chromatin was sheared with the Covaris S220 ultrasound system. For ChIP antibody reactions polyclonal rabbit antibodies were used (H3K4me1 (Abcam), H3K27Ac (Abcam), H3K27me3 (Millipore)). Washing and DNA purifications as well as size selections were performed with AMPure XP SPRI beads (Beckman Coulter). Published library construction protocol was adapted (Blecher-Gonen et al., Nature Protocols, 2013) and performed with 0.5 ng ChIP DNA. Barcodes from the NEXTflex adapter oligonucleotides kit (Bioo Scientific) were ligated to the ChIP-seq DNA fragments. Final DNA purification with additional size selection was performed with SPRI beads. ChIP-seq libraries were sequenced on the HiScanSQ/Hiseq 1000 sequencer (Illumina)