GSM7061985: t30-TCDD-AhR-ChIP; Homo sapiens; ChIP-Seq
Chromatin for ChIP was prepared as previously described(Kouwenhoven et al., 2010). ChIP assays were performed following a standard protocol (Novakovic et al., 2016) with minor modifications. On average, 0.5M keratinocytes were used in each ChIP. For histone mark H3K27ac, 2x ChIP reactions were pooled to prepare 1x ChIP-seq sample; for AHR, 4x ChIP reactions are pooled to prepare 1 ChIP-seq sample. Antibodies against H3K27ac (Diagenode #C15410174) and AHR (Santa Cruz Biotechnology, #sc-5579) were used in each ChIP assay. Resulted DNA fragments from four independent ChIP assays were purified and subjected to a ChIP-qPCR quality check. 5ng DNA fragments were pooled and proceeded on with library construction using KAPA Hyper Prep Kit according to the standard protocol. The prepared libraries were then sequenced using the NextSeq 500 (Illumina) according to standard Illumina protocols.