GSM8390284: A485 0.5uM, H3K27ac, rep2; Homo sapiens; ChIP-Seq
For spike-in normalized PerCell ChIP-seq, human Rh4 and mouse C2C12 cells were fixed in 1% formaldehyde for 10 minutes before quenching with 125 mM glycine. Cells were combined in a ratio of 3 human:1 mouse cells prior to sonication. Sonicated chromatin was immunoprecipitated with antibodies against H3K27ac (Active Motif #39133) or CTCF (CST #2899). Input and ChIP-enriched DNA was decrosslinked and purified using Qiagen MinElute PCR Purification columns prior to library preparation. ChIP and Input DNA libraries were prepared by blunt end repair using the Lucigen End-It DNA End-Repair Kit, 3' A-tailing by Klenow fragment (3'-5' exo-) (NEB), adaptor ligation by T4 DNA ligase(NEB), and size selection on an E-Gel 2% EX agarose gel. Libraries were amplified with barcoded primers for 14 cycles and isolated from unreacted primers by gel purification. Libraries were sequenced on either the HiSeq4000 or NovaSeq6000 platforms, each running in PEx150bp mode.