GSM9568102: HEK293T, ChIP-seq, WT, AM tag; Homo sapiens; ChIP-Seq
HEK293T cells transfected with pMYS-IG vector, MECOM-WT or MECOM-R750W with AM tag were fixed with 1% formaldehyde (Sigma) and then quenched with glycine. After washing and cell lysis, the chromatin was fragmented with 0.75 µl micrococcal nuclease (MNase) at 37 C for 15–20 min and nuclei were completely lysed by sonication. Chromatin was then harvested, immunoprecipitated, and DNA purified using the SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) (Cell Signaling, #9003). Anti-AM tag (active motif, catalog #91112) was used as the antibody for immunoprecipitation overnight at 4C with rotation. Add A and adapter ligation to purified DNA, the DNA fragments were amplified by PCR.ChIP-seq libraries were sequenced using Illumina Novaseq 6000 with paired-end mode.