alias: E-MTAB-6994:Input_WT_Day_Rep2_p
title: Illumina HiSeq 4000 paired end sequencing; Histone H3K27ac ChIP-seq binding regions in mouse liver
design: Histone H3K27ac ChIP-seq binding regions in mouse liver
library_construction_protocol: Male and female Reverb alpha global knockout mice and their wild-type littermate controls, aged between 9-16 weeks, were housed on a 12hr:12hr light:dark cycle, with ad libitum access to food and water. Mice were injected at ZT6 or ZT18 with 1mg/kg dexamethasone (Sigma) i.p.. One hour later, mice were culled by cervical dislocation, and livers harvested and flash frozen.  ChIP was performed using the ChIP-IT High Sensitivity kit (Active Motif) according to manufacturer instructions. A TissueRuptor (Qiagen) was used for 45s at maximum speed to disrupt liver tissue, followed by dounce homogenising for 70 strokes to release cell nuclei. Nuclear sonication was conducted with the EpiShear probe sonicator (Active Motif, 3.5mm probe) for 8 rounds of 2 minutes per sample, at 37% amplitude. 15ug sheared chromatin was incubated with anti-H3K27Ac antibody (Active Motif, Lot No: 31814008). Antibody was pulled down with magnetic protein G agarose microspheres (ReSyn Biosciences). After elution and de-crosslinking ChIP DNA was purified as per ChIP-IT High Sensitivity kit instructions. 5-10ng ChIP DNA was submitted to the University of Manchester Genomics Facility for library preparation and paired-end sequencing on the Illumina HiSeq 4000 platform.
identifiers.namespace: UNIVERSITY OF MANCHESTER
identifiers.id: E-MTAB-6994:Input_WT_Day_Rep2_p