title: GSM9266057: ChIP-Seq, human iPSC-derived microglia, untreated, control, without antibody, ChIP, rep3; Homo sapiens; ChIP-Seq
library_name: GSM9266057
library_construction_protocol: iPSC-derived microglia (iMGL) were seeded at 4 × 10⁶ cells/ml in 10 cm dishes, crosslinked with 1% formaldehyde for 10 min at RT, and quenched with 10X glycine for 5 min. Cells were washed with cold PBS, scraped in PBS + protease inhibitors, and nuclei were isolated. Chromatin was digested with micrococcal nuclease at 37 °C for 20 min and briefly sonicated to yield 150–900 bp fragments. Immunoprecipitation was performed overnight at 4 °C with IgG, or STAT1 antibodies. Complexes were captured using Protein G magnetic beads (2 h, 4 °C), washed with low/high salt buffers, and eluted. Crosslinks were reversed with 5 M NaCl and proteinase K at 65 °C for 2 h. DNA was purified via spin columns. ChIP DNA libraries were prepared using the Illumina ChIP-Seq DNA Sample Prep Kit (Illumina,IP‑102‑1001) following end-repair, A-tailing, and adapter ligation steps, followed by PCR amplification and size selection. Final libraries were purified, quantified, and quality-checked before sequencing on an Illumina platform.
identifiers.namespace: GEO
identifiers.id: GSM9266057_r1