GSM8308828: ChIP_961C_HoxBlinc_KO_IgG; Mus musculus; ChIP-Seq
10M cells were collected by centrifugation and then washed by 1×PBS. Washed cells were cross-linked by 1% formaldehyde and quenched by 125mM glycine. Crosslinked cell pellet was resuspended by ChIP lysis buffer and subjected to chromatin shearing using Bioruptor (Diagenode) with 30s on and 30s off condition. Sonicated chromatin was incubated with anti-CTCF, anti-V5, anti-H3K4me3 or anti-MLL1 antibody at 4C overnight. Prewashed Dynabeads™ Protein A or G beads (Invitrogen) were added to antibody-bounded chromatin and incubated for 4h at 4C. Antibody/Protein beads bounded chromatin were washed 5 times and reverse-crosslinked. ChIP DNA Clean & Concentrator™ (ZYMO research) was used for DNA purification. ChIP-seq library was prepared with NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB).