GSM8143846: siNT, RPA, ChIP; Homo sapiens; ChIP-Seq
Cells were crosslinked with 1% formaldehyde for 10minutes at 37°C. Crosslinking reaction was quenched with 125mM glycine for 5minutes and cells were washed 3 times with PBS. Cell pellets were resuspended in 2.5mL lysis buffer A (5mM Pipes pH 8.0, 85mM KCl, 0.5% NP40, 1mM PMSF) supplemented with protease inhibitors and incubated for 10minutes on ice. Libraries were amplified for N-1 cycles (being N the optimum Cq determined by qPCR reaction) using NEBNext High-Fidelity Polymerase (New England Biolabs, M0541). Libraries were purified with Sera-Mag Select Beads (GE Healthcare, 29343052)