GSM6730619: H3K27ac, SCN, ZT3, biol rep 1; Mus musculus; ChIP-Seq
Chromatin immunoprecipitation (ChIP) was conducted by Diagenode ChIP-seq/ChIP-qPCR Profiling service (Diagenode Cat# G02010000). Briefly, the chromatin was prepared using the True MicroChIP Kit (Diagenode Cat# C01010130). Samples were fixed with 1% FA for 8 min. Chromatin was sheared using Bioruptor® Pico sonication device (Diagenode Cat# B01060001) combined with the Bioruptor® Water cooler for 7 cycles using a 30'' [ON] 30'' [OFF] settings. Then, shearing was performed in 0.65 ml Bioruptor® Pico Microtubes (Diagenode Cat# C30010011). 500 ng of chromatin were immune-precipitated using 1 µg of each of the following Diagenode antibodies H3K4me3 (C15410003; Lot Nr. A1051D), H3K27ac (C15410196; Lot Nr. A1723-0041D) and Rabbit IgG (C15410206; Lot Nr. RIG001). Chromatin corresponding to 1% was set apart as Input. H3K4me3 and H3K27ac immune-precipitated genomic DNA along with their corresponding input samples was sent to Oxford Genomics Centre, University of Oxford for library preparation and paired-end sequencing on NovaSeq6000 platform (Illumina).