GSM8705423: A549 cells, control, INPUT; Homo sapiens; ChIP-Seq
Approximately 2 million cells were used for each ChIP sample. The cells were cross-linked with 1% formaldehyde at room temperature for 10 min. After quenching and cell lysis,chromatin was sonicated at 20% power for 3 min using a sonicator (SONICS, VCX150). For each sample, ChIP product was used for library preparation. The ChIP product was treated with End Prep Enzyme Mix for end repairing, 5' Phosphorylation and dA-tailing in one reaction, followed by ligation to adaptors with a “T” base overhang. Adaptorligated DNA was then recovered using Beads. Each sample was then amplified by PCR using P5 and P7 primers, with both primers carrying sequences which can anneal with flowcell to perform bridge PCR and P7 primer carrying index allowing for multiplexing. The PCR products were cleaned up using beads, validated using an Agilent 2100 Bioanalyzer, and quantified by Qubit 3.0 Fluorometer. Then libraries with different indices were multiplexed and loaded on an Illumina NovaSeq 6000 instrument according to manufacturer's instructions (Illumina, San Diego, CA, USA).