GSM5570047: OCI-Ly7_OCABi_OCT2_rep1; Homo sapiens; ChIP-Seq
10 million cells were fixed in 1% formaldehyde for 10 min at room temperature, and fragmentation of fixed chromatin was obtained from isolated nuclei by sonication. To enrich for short chromatin fragments (200 - 700 bp), 5μg of antibody was added to the chromatin lysate and incubated overnight at 4 °C. The following day, Dynabeads protein A was added and incubated with rotation at 4 °C for 1.5 hours. Enriched DNA was isolated through extensive wash steps, subsequent reverse cross-linking, and purification using DNA Clean & Concentrator Kit. ChIP-seq libraries were prepared from 2-5 ng ChIP DNA. After end-repair, A-tailing, and ligation with barcodes, barcoded DNA was amplified by 12 cycles of PCR using Phusion® High-Fidelity DNA Polymerase. 50bp single-end.