title: GSM8438850: F36P AM-ERG H3K27ac ChIP; Homo sapiens; ChIP-Seq
library_construction_protocol: All samples were carried out using SimpleChIP® Enzymatic Chromatin IP Kit (CST, 9003) following the manufacturer's instructions. 10^7 cultured F36P cells per sample were fixed with 1% formaldehyde and then quenched with glycine. After washing and cell lysis, the chromatin was fragmented with 0.7 µl of micrococcal nuclease (MNase) at 37oC for 20 min and nuclei were completely lysed by sonication. The 10 µg of chromatin in each reaction was incubated with 10 µl of Acetyl-Histone H3 (Lys27) antibody overnight at 4oC with rotation. Immunoprecipitation was performed with protein G magnetic beads. Following elution, reverse-crosslinks, and purification, DNA was used for further sequencing. The DNA fragments were repaired, A-tailed and further ligated with Illumina adapter. The final DNA library was obtained by size selection and PCR amplification.