GSM5941001: RAW264.7 HDAC3KO-Ace-2; Mus musculus; ChIP-Seq
Lysates were clarified from sonicated nuclei and H3K27ac-DNA complexes or IgG-DNA complexes were isolated with antibody. Library preparation for ChIP-seq was using KAPA HyperPlus Kit (KAPA Biosystems, #KK8514) according to manufacturer's protocol. Briefly, 50 μl sample was proceed to End Repair and A-tailing at 65 ℃ for 30 min. Different adapter was added into each sample and incubated at 20 ℃ for 15 min. After ligation, DNA was bound to the beads and washed by 80% ethanol for 3 times, followed by 7-8 cycles PCR. The DNA was performed 1x bead-based cleanup again and the product was ready for sequencing which was done by company (Novogene).