title: GSM6463421: K562 Con Input ChIP-seq; Homo sapiens; ChIP-Seq
library_construction_protocol: Nuclei were digested with 100 units of MNase at 37°C for 15 min. Mono or di-nucleosomes sized chromatin was incubated with antibodies for 3 h at 4°C and recovered with protein G or A agarose beads. ChIP DNA (10 ng for K27ac, Input, 0.5 ng for GATA1, TAL1, LDB1) was processed using NEBNext ChIP-seq library (New England Biolabs) with manufacturer's instructions. The ends of ChIP DNA were repaired by NEBNext Ultra II End Prep enzyme mix and ligated with NEBNext adaptors. Adaptor-ligated DNA was selected at 200 bp size using NEBNext sample purification bead and amplified using the adaptor primers. Amplification cycles were 7 cycles for H3K27ac and input and 12 cycles for GATA1, TAL1 and LDB1. The fragments are purified using NEBNext sample purification beads.