HiSeq X Ten sequencing; The H3K4me3 pattern of mouse renal cortex tissues treated with dexmedetomidine against lipopolysaccharide-induced sepsis
The H3K4me3 pattern of mouse renal cortex tissues treated with dexmedetomidine against lipopolysaccharide-induced sepsis
Mouse kidney tissues were removed capsule, and cut into small pieces in cold PBS, and added RIPA buffer (0.5% SDS) on ice for 30 min, then centrifuged at 13000 rpm for 15 min. The supernatant was harvested and treated by sonication to break genomic DNA into 200-500 bp. 10% were stored as input and the rest of supernatant was quantified the equal DNA concentration, then incubated with 1 μg IP grade antibodies of H3K4me3 (Activemotif, USA) 4 °C overnight. Protein-A beads were incubated next day at 4 °C for target protein pull down. template DNA was repaired to 3'-dA overhang and added the ligated adapter. The DNA library was eliminated the unligated adapters and selected the appropriate size for sequence using an Illumina Hiseq 2000 platform.