GSM2224623: downstream CTCF viewpoint in the liver; Mus musculus; OTHER
4C templates was prepared as described previously (Simonis et al., 2009). Adult mouse hearts and liver were dissociated and homogenized in ice-cold PBS with 10%FCS to obtain single cell suspension. Chromatin was cross-linked with 2% formaldehyde in 40ml PBS with 10% FCS for 10 min at room temperature, nuclei were isolated in 25ml cold lysis buffer for 1h, and cross-linked DNA was digested with DpnII. Digestion was followed by proximity ligation, removal of cross-links, a secondary restriction digestion with Csp6I and a second proximity ligation. 200 ng of the resulting 4C template was used for the subsequent PCR reaction using Expand Long Template Polymerase (Roche). The PCR products were purified using the High Pure PCR Product Purification Kit (Roche) and the QIAquick PCR purification kit (Qiagen).
GEO Accession
GSM2224623