GSM9103959: HS02 DMSO A2; Homo sapiens; ChIP-Seq
Flasks were crosslinked for using 10% formaldehyde for 10 min at room temperature and quenched with 25M glycine for 5 min. Nuclear extracted was performed and DNA was digested with MNase (50units/10^6 cells) followed by sonication (30% amplitude, 6 cycles, 10 sec ON, 30 sec OFF). Immunoprecipitation was performed overnight with rabbit anti-BRD4 antibodies (#BL-149-2H5, Fortis) conjugated to Dynabeads Protein A (ThermoFisher). After elution, DNA was reverse crosslinked, thenRNase and proteinase K treated. DNA was extracted using the ChIP DNA Clean & Concentrator Kit (Zymo) and analyzed using Nanodrop8000 and Tapestation 4150 bioanalyzer (Agilent). Samples were sent to Novogene for library construction. From the quality analysis report "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. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. The library was then sequenced using the Illumina NovaSeq platform (12G raw data per sample; 150bp, paired end reads).