Adult mouse hearts and livers were isolated in ice cold PBS. Single cell suspensions were obtained by dissociation of tissue with IKA Ultra Turrax T5 FU, followed by dounce homogenization. Chromatin was cross-linked with 2% formaldehyde in PBS with 10% FCS for 10 min at room temperature, nuclei were isolated and cross-linked DNA was digested with a primary restriction enzyme recognizing a 4 bp restriction site (DpnII), followed by proximity ligation. Cross-links were removed and a secondary restriction enzyme digestion (Csp6I), followed again by proximity ligation. 200 ng of the resulting 4C template was used for subsequent PCR reactions using Expand Long Template Polymerase (Roche). PCR products were purified using the High Pure PCR Product Purification Kit (Roche, 11732676001).
GEO Accession
GSM3964183