GSM4462232: SUM159_ZNF446_ChIP_rep2; Homo sapiens; ChIP-Seq
Cells were crosslinked with 1% formaldehyde for 15                     min at 37°C. Crosslinking was quenched with 0.125 M glycine for 5 min at 4°C,                     followed by multiple washes with cold PBS. Nuclei were harvested via hypotonic                     lysis and subjected to sonication in a lysis buffer containing 1% SDS. Sheared                     chromatin lysates were then pre-cleared with Protein A/G agarose beads for 1                     hour, followed by immunoprecipitation with appropriate antibody overnight at                     4°C. Protein A/G agarose beads blocked with BSA were added to lysates for 2                     hours the next day, followed by extensive washes. Protein/DNA complexes were                     eluted from the beads with 100 mM NaHCO3 and 1% SDS, and de-crosslinking was                     carried out overnight at 65°C. RNA and protein were digested with RNase A and                     Proteinase K, respectively. DNA was purified using phenol/chloroform/isoamyl                     alcohol (25:24:1) and precipitated overnight at -20°C. Libraries of ChIP DNA                     were prepared using the KAPA Hyper Prep Kit (KAPA Biosystems) according to                     manufacturer's instructions, and samples were multiplexed with Illumina adapters                     (NEB).