GSM1437734
GSM1437734: RORalpha Liver ZT22 ChIP-seq; Mus musculus; ChIP-Seq
ChIP experiments were performed as previously described (Feng et al., 2011) with minor changes. Mouse liver was harvested, minced and cross-linked in 1% formaldehyde for 20min, followed by quenching with 1/20 volume of 2.5M glycine solution for 5 minutes, and two washes with 1× PBS. Nuclear extracts were prepared by Dounce homogenization in ChIP buffer (50mM Tris-HCl pH 7.5, 140mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% NaDOC). Chromatin fragmentation was performed by sonication in lysis buffer (50mM Tris-HCl pH 8.0, 1%SDS, 10mM EDTA), using the Bioruptor (Diagenode). Proteins were immunoprecipitated in ChIP buffer, and cross-linking was reversed overnight at 65°C in lysis buffer (50mM Tris-HCL, 10mM EDTA, 1% SDS, pH 8), and DNA isolated using phenol/chloroform/isoamyl alcohol. DNA was amplified according to ChIP Sequencing Sample Preparation Guide provided by Illumina, using adaptor oligo and primers from Illumina, enzymes from New England Biolabs and PCR Purification Kit and MinElute Kit from Qiagen.
GEO
GSM1437734
GEO Accession
GSM1437734