GSM8012583: Embryo, 14-15 stage, H3K27ac E7, ChIP-seq, rep1; Drosophila melanogaster; ChIP-Seq
Stage 14-15 embryos from a line containing E10::GFP and 7::DsRed transgenes (Preger-Ben Noon, Davis, and Stern 2016) were cross-linked, dissociated and isolated nuclei were immunostained with anti-GFP and anti-DsRed antibodies and the appropriate secondary antibodies. The E10::GFP and 7::DsRed nuclei, which constitute the majority of svb expressing nuclei, were then isolated by fluorescence activated cell sorter (FACS). Chromatin from 250,000 nuclei of each cell sub-populations (n=3) was isolated and used for ChIP with anti-H3K27ac and anti-H3 antibodies (Abcam) using the iDeal ChIP-seq kit (Diagenode). Libraries were prepared using the Ovation Ultralow V2 DNA-Seq library preparation kit (NuGen) according to the manufacturer instructions. Libraries were sequenced on a NextSeq 550 system (Illumina) with 50 bp SE reads.