GSM9282474: H9 cell line, IKZF3 ChIP; Homo sapiens; ChIP-Seq
fixed 5-10 million cells per IP were lysed in prepared Buffer A for 10 min at room temperature. After lysing, nuclei were digested with prepared Buffer B and Micrococcal Nuclease for 20 min at 37 °C. After adding 50 mM EDTA to stop digestion, nuclei were collected (16,000× g, 4°C, 1 min) and sonicated with two pulses of 40 seconds each and 30 s incubation on wet ice. 10 µg chromatin was incubated with primary antibody and equal amount of IgG antibody overnight. Chromatin was then incubated with 30 µL Dynabeads Protein G (Invitrogen #10004D) for 4 hours and washed with low-salt and high-salt washing buffer once. Elution and de-crosslinking were performed overnight at 65 °C. Eluted samples were purified by QIAquickTM PCR Purification Kit (Qiagen #28104) NEBNext ChIP-Seq Library Prep Master Mix Set for Illumina (NEB #E6240) was used according to the manual's instructions. Library quality was determined by TapeStation prior to being sequenced on the Illumina NovaSeq X plus platform.